具有非FcγR依赖性激动活性的肿瘤坏死因子(TNF)受体超家族(TNFRSF)受体-激活抗体融合蛋白(具有非FcγR依赖性激动活性的TNFRSF受体-激活抗体融合蛋白；TRAAFFIAA)
阅读说明：本技术 具有非FcγR依赖性激动活性的肿瘤坏死因子(TNF)受体超家族(TNFRSF)受体-激活抗体融合蛋白(具有非FcγR依赖性激动活性的TNFRSF受体-激活抗体融合蛋白；TRAAFFIAA) (Tumor Necrosis Factor (TNF) receptor superfamily (TNFRSF) receptor-activating antibody fusion proteins with non-Fc γ R dependent agonistic activity (TNFRSF receptor-activating antibody fusion proteins) 是由 安德烈亚斯·拜尔哈克 尤利亚妮·库姆斯 约翰内斯·内尔克 哈拉尔德·瓦赞特 于 2018-12-20 设计创作，主要内容包括：本发明涉及具有非FcγR依赖性激动活性的肿瘤坏死因子(TNF)受体超家族(TNFRSF)受体-激活抗体融合蛋白,及其组合物和方法。(The present invention relates to Tumor Necrosis Factor (TNF) receptor superfamily (TNFRSF) receptor-activating antibody fusion proteins having non-Fc γ R dependent agonistic activity, and compositions and methods thereof.)
技术领域
本发明涉及具有非FcγR依赖性激动活性的肿瘤坏死因子(TNF)受体超家族(TNFRSF)受体-激活抗体融合蛋白(缩写为TRAAFFIAA)，及其组合物和方法。
背景技术
肿瘤坏死因子(TNF)受体超家族的受体及其激活
大多数肿瘤坏死因子(TNF)受体超家族(TNFRSF)的受体是1型或3型跨膜蛋白，其胞外结构域具有1-6个结构保守的富含半胱氨酸的结构域(CRD)。另外，存在一些分泌的或GPI锚定的受体分子，由于它们具有CRD，因此被认为是TNFRSF的成员。除上述结构之外，TNFRSF的所有受体没有其他共有的结构特征。根据结构和功能的相似性，可以在TNFRSF中定义三个亚组(图1)：
1.死亡受体亚组中的TNFRSF受体，由其在胞内结构域中结构保守的蛋白质-蛋白质相互作用结构域定义，即它们的“死亡结构域”(DD)，从而命名。这些受体中的一部分而非全部能诱导细胞凋亡或坏死，但也可以触发其他作用/信号通路。例如CD95(也称为Fas或Apo1)、TRAILR1(DR4)和TRAILR2(DR5)。
2.TRAF互作亚组中的TNFRSF受体，其不具有DD但具有短的氨基酸序列基序，通过基序募集TNF受体相关因子(TRAF)家族的衔接蛋白。借助于这些TRAF蛋白，该TNFRSF亚组的受体可以激活各种信号通路。例如TNFR2、CD40、4-1BB、OX40、CD27、Fn14和BaffR。
3.TNFRSF的诱骗受体，其不具有胞内结构域，并被分泌出来且被质膜外表面的GPI基团锚定。这些TNFRSF受体自身不传递信号，但控制亚组1和2的某些受体的活性。
通常，TNFRSF的信号可转导受体通过与TNF超家族(TNFSF)的配体相互作用而自然激活。TNFSF配体的分配受C端保守域——TNF同源域(THD)的支配。通过其THD，TNFSF配体形成同三聚体分子，在少数情况下还形成异三聚体分子。除了分泌的LTalpha同型三聚体外，TNF家族的所有配体最初都作为II型膜蛋白表达。因此，THD具有胞外定位。但是，通过THD和TNFSF配体的跨膜结构域之间的蛋白水解过程，在这些情况下也可以形成可溶性三聚体配体分子(图1)。单个TNFRSF受体分子被招募到由配体三聚体的相邻原聚体之间形成的接触表面区域中。因此，一个TNFSF配体分子结合TNFRSF的三个受体；这适用于可溶性和膜结合的TNFSF配体分子。目前所知，膜结合的TNFSF配体的结合总是导致强烈的受体活化。但是，可溶性TNFSF配体结合后胞内信号传导途径的强劲有效激活的程度，取决于特定的TNFRSF受体。因此，TNFRSF受体如TNFR1、GITR、LTbR或DR3被可溶性配体三聚体的结合而强烈激活，而TNFRSF受体如TNFR2、Fn14、TRAILR1、TRAILR2、CD95、OX40、CD27、4-1BB和CD40不能或者极低地受可溶性配体三聚体刺激。在这些情况下，所述可溶性TNF配体甚至可以充当其膜结合形式的竞争性抑制剂。为什么第二组的TNFRSF受体被其膜结合配体的刺激作用比被其可溶性配体分子的刺激作用要好得多的问题尚未得到完全解决。然而，目前认为三聚体配体-受体复合物不足以刺激大多数信号传导途径，并且这些复合物中的至少两种必须以二级方式相互作用以用于有效地启动信号传导。据此，TNFRSF的某些受体显示出其具有尽管弱，但不依赖配体的自体亲和力。与可溶性配体相比，膜结合的配体三聚体的膜定位导致空间预取向、扩散降低，并且相互作用配偶体在TNFRSF受体表达细胞和TNFSF配体表达细胞之间的接触区局部浓度非常高。因此，在结合膜结合的配体的情况下，最初形成的三聚体配体-受体复合物都可能发生自发性二次聚集。然而，在结合可溶性配体三聚体的情况下，第二组的TNFRSF受体的自体亲和力可能太弱以至于不能确保所述二次聚集。根据该模型，与膜结合配体类似，分别包含两个或更多个三聚体配体结构域的可溶性融合蛋白和TNFSF配体复合物可以激活第二组的TNFRSF受体。此外，还显示仅将三聚体可溶性TNFSF配体仅在细胞或其他表面上的锚定就足以激活第二组的TNFRSF受体(参见Wajant et al.,Cancer Lett.2013May 28；332(2):163-74.)。
激活TNFRSF的受体的第二种可能性是使用抗体。考虑到各种临床概念，特别是旨在激活TNFRSF的细胞死亡诱导或免疫刺激受体的涉及肿瘤疾病的治疗，使用抗体是非常重要的。因此，在临床研究中正在测试或已经测试了靶向TNFRSF受体TRAILR2、CD40、Fn14、CD27、OX40和4-1BB的抗体。在这一方面，长期以来一直忽略了抗TNFRSF受体特异性抗体何时以及在什么情况下以受体刺激方式起作用的问题。通常认为这主要是抗体具体识别的表位的问题。尤其是过去几年，在这方面的研究表明，与可溶性TNFSF配体相似，主要是所述分子的化合价和呈递方式决定了是否发生受体活化，即所述抗体是否作为激动剂。因此，对于识别TNFRSF受体的各种二价IgG1和IgG2抗体(其未被可溶性配体充分刺激)，发现只有当它们被第二抗体或G蛋白以二级方式交联时，或者这些抗体可以同时与细胞结合的Fcγ受体(FcγRs)结合时，其才能有效地充当激动剂(参见Wajant 2015,Cell DeathDiffer.2015Nov；22(11):1727-41和图2-3)。
这可能导致看似自相矛盾的情况，即阻断配体结合的拮抗抗体在与FcγRs结合后充当强激动剂，即不是识别的表位，而主要是抗体与其抗原结合的形式决定抗体的作用(参见Trebing et al.,MAbs.2014 Jan-Feb；6(1):297-308的图3D，出于所有目的通过引用整体并入本文)。此外，发明人观察到已经被可溶性配体有效激活的、识别TNFRSF受体的IgG抗体通常也以激动剂的方式作用，而没有进一步的交联或结合FcγR(图2，参见包含有关αTNF1和αLTR的实施例小组)。
对识别(不受可溶性TNF配体和抗体刺激的)TNFRSF受体的二价IgG抗体的激动活性的要求复杂，甚至阻碍其在临床方法中的使用。由于所述抗TNFRSF受体抗体结合FcγR不仅导致TNFRSF受体的活化，而且刺激Fcγ受体，所以可能发生不希望发生的反应，这使临床应用复杂化或甚至排除了其临床用途。与活化的FcγR的结合可能，例如导致ADCC、CDC或吞噬作用介导的表达TNFRSF受体的细胞的去除/破坏。为了防止这种情况，可以使用更强烈或更优选地与抑制性FcγR、CD32B相互作用的IgG突变体。但是，CD32B的免疫抑制活性也可能造成困扰。然而，更具体地说，不能保证在表达靶TNFRSF受体的靶细胞附近总能找到足够数量的表达CD32B的细胞。尽管IgM抗体和IgG3抗体极易发生自体聚集，但由于它们的高亲和力，在不存在FcγR结合的情况下可以以激动剂的方式作用——类似于以二级方式寡聚化的可溶性配体三聚体——与IgG1、IgG2和IgG4抗体相比，所述抗体的开发更加费力且更不成熟。
因此，存在靶向TNFRSF受体的改良剂的需求。具体地，期望获得有效且更可靠地靶向TNFRSF的受体，并且与现有的TNFRSF靶向制剂相比更不可能具有有害副作用的制剂。
本发明解决了上述问题并满足了本领域中的上述需求。
如本发明所述的抗TNFRSF受体抗体融合蛋白与细胞结合的结构或与胞外基质的结构结合，所述结合的方式不依赖于抗体部分的抗原结合结构域以及可能存在的天然或突变的Fc结构域。意外的是，发明人发现所述融合蛋白，即包含抗TNFRSF受体抗体(或其抗原结合部分)的融合蛋白，在与靶细胞结构或胞外基质相互作用后充当强激动剂，即不依赖于FcγR结合的TNFRSF受体刺激方式(参见，例如图4-12作为非限制性实施例)。
上述发现是有利的，有如下理由：例如，不依赖FcγR的刺激是有利的，因为它不限于表达FcγR的情形。另外，这也是有利的，因为可以避免由于FcγR的刺激而引起的副作用。此类副作用包括ADCC、CDC或吞噬作用去除/破坏表达TNFRSF受体的细胞，以及FcγRCD32B的免疫抑制活性。
因此，一方面，本发明描述了抗体融合蛋白的形式，其允许有效刺激TNFRSF的受体而不结合FcγR。在适当选择与抗TNFRSF受体抗体融合的结构域后，如本发明所述的具有不依赖FcγR的激动活性的，肿瘤坏死因子(TNF)受体超家族(TNFRSF)受体激活抗体融合蛋白(TRAAFFIAA)也可以是双功能的和/或具有前药性质。如本文所述，“双功能的”是指细胞表面或胞外基质的结构(例如细胞表面或胞外基质的蛋白质)，当被能以不依赖FcγR的方式结合所述结构的结构域结合时，其被抑制或刺激。TRAAFFIAA是双功能的，是由于它不仅激活TNFRSF受体信号传导，而且还调节靶分子结构的活性。如本文所述，前药被肿瘤相关因子(例如酶，缺氧)激活。例如，靶向肿瘤抗原的TRAAFFiAA仅在肿瘤中获得激动活性，并如前药一般作用。
从本发明人的工作中可立即显而易见的是，在TRAAFFIAA中使用经适当选择的固定化结构域，可以在体内获得限于靶结构的局部激动效应。从而有可能避免可能限制治疗的全身性副作用。因此，TRAAFFIAA不仅可以以不依赖FcγR的方式有利地使用抗体刺激TNFRSF受体，而且还开拓了新的应用领域，包括临床应用。
因此，本发明包括以下优选实施方案：
1.一种融合蛋白，包含：
i)抗TNFRSF受体抗体或其抗原结合部分，和
ii)能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域。
2.如第1项所述的融合蛋白，其中，所述结构为表达TNFRSF受体的细胞的细胞表面结构，或与表达TNFRSF受体的细胞相邻的细胞的细胞表面结构。
3.如第1项所述的融合蛋白，其中，所述结构为表达TNFRSF受体的细胞的胞外基质的结构，或与表达TNFRSF受体的细胞相邻的细胞的胞外基质的结构。
4.如第2项所述的融合蛋白，其中，所述结构为细胞表面蛋白或糖，优选为细胞表面蛋白。
5.如第3项所述的融合蛋白，其中，所述结构为胞外基质蛋白或糖，优选为胞外基质蛋白。
6.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分为单克隆。
7.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分为全长抗体、Fab2片段、Fab片段或其中的VH和VL已被TNFRSF受体特异性scFv片段取代的抗体。
8.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分不包含Fc结构域。
9.如第1-7项中任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分为抗体变体，所述抗体变体与一种或多种FcγR类型的结合能力降低；优选为包含N297A突变的全长抗体。
10.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分为IgG1、IgG2或IgG4抗体或其抗原结合部分。
11.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分为二价抗体或其抗原结合部分。
12.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分选自抗TNFR2抗体或其抗原结合部分、抗CD40抗体或其抗原结合部分、抗CD95抗体或其抗原结合部分、抗Fn14抗体或其抗原结合部分、抗TRAILR2抗体或其抗原结合部分、抗TRAILR1抗体或其抗原结合部分、抗CD27抗体或其抗原结合部分、抗OX40抗体或其抗原结合部分、抗4-1BB抗体或其抗原结合部分、抗BaffR抗体或其抗原结合部分、抗TACI抗体或其抗原结合部分、或者抗BCMA抗体或其抗原结合部分组成的组。
13.如前述任一项所述的融合蛋白，其中，如i)所述的抗体或其抗原结合部分选自抗TNFR2抗体或其抗原结合部分、抗CD40抗体或其抗原结合部分、抗CD95抗体或其抗原结合部分、或者抗Fn14抗体或其抗原结合部分组成的组。
14.如前述任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述的scTNFSF配体优选为TNFR2、GITR、4-1BB、BaffR、TACI、CD40、Fn14或OX40的配体。
15.如前述任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或抗体的抗原结合部分，所述抗体或抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构。
16.如第15项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的所述结构的抗体或抗原结合部分为能结合PD-L1的检查点抑制剂抗体或其抗原结合部分，优选为Avelumab，或其PD-L1结合部分。
17.如第15项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的结构的抗体或抗原结合部分为抗CD27抗体或其抗原结合部分。
18.如第15-17项中任一项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分为结合一种或多种FcγR类型能力降低的抗原结合部分，所述抗原结合部分优选为Fab2片段、scFv片段或Fab片段，更优选为scFv片段或Fab片段。
19.如第15-18项中任一项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的结构的抗原结合部分为Fab片段。
20.如第15-18项中任一项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的结构的抗原结合部分为scFv片段。
21.如第15-20项中任一项所述的融合蛋白，其中，所述能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分或其抗原结合部分选自抗CD20抗原结合部分、抗CD70抗原结合部分、抗CD19抗原结合部分、抗EGFR抗原结合部分、抗Her2抗原结合部分、抗Fn14抗原结合部分、抗CD40L抗原结合部分、或者抗PD1L抗原结合部分组成的组。
22.如前述任一项所述的融合蛋白，其中，如ii)所述的结构域不包含Fc结构域。
23.如第1-14项中任一项所述的融合蛋白，其中，如ii)所述的结构域不包含抗体或其抗原结合片段。
24.如第1-14项或第23项中任一项所述的融合蛋白，其中，如ii)所述的结构域与细胞表面的所述结构和/或胞外基质的所述结构的所述结合是不依赖抗原的。
25.如第1-16项和第18-24项中任一项所述的融合蛋白，其中，如ii)所述的结构域不包含抗TNFRSF受体抗体或其抗原结合部分。
26.如前述任一项所述的融合蛋白，其中，如ii)所述的结构域包含干扰素或白介素结构域。
27.如前述任一项所述的融合蛋白，其中，如ii)所述的结构域包含，并优选由IL-2、IL-4、IL-10、IFNα、IFNβ或IFNγ组成；或者包含，并优选由IL-2、IL-4、IL-10、IFNα、IFNβ或IFNγ的变体组成，所述变体能以不依赖FcγR的方式结合细胞表面的所述结构和/或胞外基质的所述结构；其中所述IL-2、IL-4、IL-10、IFNα、IFNβ或IFNγ优选为人IL-2、人IL-4、人IL-10、人IFNα、人IFNβ或人IFNγ。
28.如前述任一项所述的融合蛋白，其中，与包含如i)所述的抗TNFRSF受体抗体或其抗原结合部分但不包含如ii)所述的结构域的蛋白相比，所述融合蛋白能增强对所述TNFRSF受体的刺激。
29.抗TNFRSF受体抗体或其抗原结合部分的融合蛋白，其特征在于，所述融合蛋白能以不依赖抗原和不依赖FcγR的方式结合所述细胞表面的结构或所述胞外基质的结构。
30.如第29项所述的融合蛋白，其与所述结构结合后能增强对TNFRSF受体的刺激。
31.如第29项或第30项所述的融合蛋白，所述融合蛋白能结合TNFRSF受体TNFR2、CD40、CD95、Fn14、TRAILR2、TRAILR1、CD27、OX40、4-1BB、BaffR、TACI或BCMA。
32.如第29-31项中任一项所述的融合蛋白，其中，所述融合蛋白为包含能结合细胞表面的所述结构或胞外基质的所述结构的scFv结构域的融合物，或者包含scTNFSF配体或其他能结合细胞表面的所述结构或胞外基质的所述结构的蛋白质结构域的融合物。
33.如第29-32项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为Fab2或Fab片段。
34.如前述任一项所述的融合蛋白，其中，所述融合蛋白选自包含如SEQ ID NO:19和23所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:21和23所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:22和23所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:24和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:25和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:26和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:28和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:29和31所示的氨基酸序列的融合蛋白、包含如SEQID NO:30和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:32和36所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:34和36所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:35和36所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:118和23所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:119和36所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:120和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:121和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:122和31所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:123和124所示的氨基酸序列的融合蛋白、包含如SEQ ID NO:125和36所示的氨基酸序列的融合蛋白。
35.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为抗CD40抗体或其抗原结合部分；所述抗CD40抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:37所示的氨基酸序列的CDR1、如SEQ ID NO:38所示的氨基酸序列的CDR2和如LDY所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:39所示的氨基酸序列的CDR1、如SEQ ID NO:40所示的氨基酸序列的CDR2和如SEQ ID NO:41所示的氨基酸序列的CDR3。
36.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为抗CD95抗体或其抗原结合部分；所述抗CD95抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:42所示的氨基酸序列的CDR1、如SEQ ID NO:43所示的氨基酸序列的CDR2和如SEQ ID NO:44所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ IDNO:45所示的氨基酸序列的CDR1、如SEQ ID NO:46所示的氨基酸序列的CDR2和如SEQ IDNO:47所示的氨基酸序列的CDR3。
37.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为抗DR5抗体或其抗原结合部分；所述抗DR5抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:48所示的氨基酸序列的CDR1、如SEQ ID NO:49所示的氨基酸序列的CDR2和如SEQ ID NO:50所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ IDNO:51所示的氨基酸序列的CDR1、如SEQ ID NO:52所示的氨基酸序列的CDR2和如SEQ IDNO:53所示的氨基酸序列的CDR3。
38.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是抗Fn14抗体或其抗原结合部分；所述抗Fn14抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:54所示的氨基酸序列的CDR1、如SEQ ID NO:55所示的氨基酸序列的CDR2和如SEQ ID NO:56所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ IDNO:57所示的氨基酸序列的CDR1、如SEQ ID NO:58所示的氨基酸序列的CDR2和如SEQ IDNO:59所示的氨基酸序列的CDR3。
39.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是抗Fn14抗体或其抗原结合部分；所述抗Fn14抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:84所示的氨基酸序列的CDR1、如SEQ ID NO:85所示的氨基酸序列的CDR2和如SEQ ID NO:86所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ IDNO:87所示的氨基酸序列的CDR1、如SEQ ID NO:88所示的氨基酸序列的CDR2和如SEQ IDNO:89所示的氨基酸序列的CDR3。
40.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是包含重链的抗TNFR2抗体或其抗原结合部分；所述抗TNFR2抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:60所示的氨基酸序列的CDR1、如SEQ ID NO:61所示的氨基酸序列的CDR2和如SEQ ID NO:62所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:63所示的氨基酸序列的CDR1、如SEQ ID NO:64所示的氨基酸序列的CDR2和如SEQ ID NO:65所示的氨基酸序列的CDR3。
41.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是抗4-1BB抗体或其抗原结合部分；所述抗4-1BB抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:126所示的氨基酸序列的CDR1、如SEQ ID NO:127所示的氨基酸序列的CDR2和如SEQ ID NO:128所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:129所示的氨基酸序列的CDR1、如SEQ ID NO:130所示的氨基酸序列的CDR2和如SEQ ID NO:131所示的氨基酸序列的CDR3。
42.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为HBBK4或其4-1BB结合部分。
43.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是抗4-1BB抗体或其抗原结合部分；所述抗4-1BB抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:132所示的氨基酸序列的CDR1、如SEQ ID NO:133所示的氨基酸序列的CDR2和如SEQ ID NO:134所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:135所示的氨基酸序列的CDR1、如SEQ ID NO:136所示的氨基酸序列的CDR2和如SEQ ID NO:137所示的氨基酸序列的CDR3。
44.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为urelumab或其4-1BB结合部分。
45.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分是抗4-1BB抗体或其抗原结合部分；所述抗4-1BB抗体或其抗原结合部分包含重链，所述重链包含如SEQ ID NO:138所示的氨基酸序列的CDR1、如SEQ ID NO:139所示的氨基酸序列的CDR2和如SEQ ID NO:140所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:141所示的氨基酸序列的CDR1、如SEQ ID NO:142所示的氨基酸序列的CDR2和如SEQ ID NO:143所示的氨基酸序列的CDR3。
46.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为utomilumab或其4-1BB结合部分。
47.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第35项所述的抗TNFRSF受体抗体交叉竞争结合CD40的抗体或其抗原结合部分。
48.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第36项所述的抗TNFRSF受体抗体交叉竞争结合CD95的抗体或其抗原结合部分。
49.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第37项所述的抗TNFRSF受体抗体交叉竞争结合DR5的抗体或其抗原结合部分。
50.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第38项所述的抗TNFRSF受体抗体交叉竞争结合Fn14的抗体或其抗原结合部分。
51.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第39项所述的抗TNFRSF受体抗体交叉竞争结合Fn14的抗体或其抗原结合部分。
52.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第40项所述的抗TNFRSF受体抗体交叉竞争结合TNFR2的抗体或其抗原结合部分。
53.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第41项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
54.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第42项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
55.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第43项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
56.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第44项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
57.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第45项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
58.如第1-33项中任一项所述的融合蛋白，其中，所述抗TNFRSF受体抗体或其抗原结合部分为能与第46项所述的抗TNFRSF受体抗体交叉竞争结合4-1BB的抗体或其抗原结合部分。
59.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或其抗原结合部分，所述抗体或其抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中所述抗体或其抗原结合部分为抗CD20抗体或其抗原结合部分，其包含重链，所述重链包含如SEQ ID NO:66所示的氨基酸序列的CDR1、如SEQ ID NO:67所示的氨基酸序列的CDR2和如SEQ ID NO:68所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:69所示的氨基酸序列的CDR1、如SEQ ID NO:70所示的氨基酸序列的CDR2和如SEQ ID NO:71所示的氨基酸序列的CDR3。
60.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或其抗原结合部分，所述抗体或其抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中所述抗体或其抗原结合部分为抗CD19抗体或其抗原结合部分，其包含重链，所述重链包含如SEQ ID NO:72所示的氨基酸序列的CDR1、如SEQ ID NO:73所示的氨基酸序列的CDR2和如SEQ ID NO:74所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:75所示的氨基酸序列的CDR1、如SEQ ID NO:76所示的氨基酸序列的CDR2和如SEQ ID NO:77所示的氨基酸序列的CDR3。
61.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或其抗原结合部分，所述抗体或其抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中所述抗体或其抗原结合部分为抗CD70抗体或其抗原结合部分，其包含重链，所述重链包含如SEQ ID NO:78所示的氨基酸序列的CDR1、如SEQ ID NO:79所示的氨基酸序列的CDR2和如SEQ ID NO:80所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:81所示的氨基酸序列的CDR1、如SEQ ID NO:82所示的氨基酸序列的CDR2和如SEQ ID NO:83所示的氨基酸序列的CDR3。
62.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或其抗原结合部分，所述抗体或其抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中所述抗体或其抗原结合部分为抗CD70抗体或其抗原结合部分，其包含重链，所述重链包含如SEQ ID NO:90所示的氨基酸序列的CDR1、如SEQ ID NO:91所示的氨基酸序列的CDR2和如SEQ ID NO:92所示的氨基酸序列的CDR3；包含轻链，所述轻链包含如SEQ ID NO:93所示的氨基酸序列的CDR1、如SEQ ID NO:94所示的氨基酸序列的CDR2和如SEQ ID NO:95所示的氨基酸序列的CDR3。
63.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体或其抗原结合部分，所述抗体或其抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中所述抗体或其抗原结合部分为a)能与如第59项所述的结合CD20的抗CD20抗体交叉竞争的抗CD20抗体或其抗原结合部分，或b)能与如第60项所述的结合CD19的抗CD19抗体交叉竞争的抗CD19抗体或其抗原结合部分，或c)能与如第61项所述的结合CD70的抗CD70抗体交叉竞争的抗CD70抗体或其抗原结合部分，或d)能与如第62项所述的结合CD70的抗CD70抗体交叉竞争的抗CD70抗体或其抗原结合部分。
64.如第1-28项和第35-63项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体的抗原结合部分，所述抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分为包含如SEQ ID NO:96所示的氨基酸序列的抗CD20 scFv片段，或包含与如SEQ IDNO:96所示的氨基酸序列至少90％同一性的抗CD20 scFv片段。
65.如第1-28项和第35-63项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体的抗原结合部分，所述抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分为包含如SEQ ID NO:98所示的氨基酸序列的抗CD19 scFv片段，或包含与如SEQ IDNO:98所示的氨基酸序列至少90％同一性的抗CD19 scFv片段。
66.如第1-28项和第35-63项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体的抗原结合部分，所述抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分为包含如SEQ ID NO:144所示的氨基酸序列的抗CD70 scFv片段，或包含与如SEQ IDNO:144所示的氨基酸序列至少90％同一性的抗CD70 scFv片段。
67.如第1-28项和第35-63项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含抗体的抗原结合部分，所述抗原结合部分能结合细胞表面的所述结构和/或胞外基质的所述结构，并且其中能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分为包含如SEQ ID NO:145所示的氨基酸序列的抗CD70 scFv片段，或包含与如SEQ IDNO:145所示的氨基酸序列至少90％同一性的抗CD70 scFv片段。
68.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述scTNFSF配体为TNFR2的配体，所述配体为具有如SEQ IDNO:102所示的氨基酸序列或与其具有至少90％同一性的scTNF80。
69.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述scTNFSF配体为具有如SEQ ID NO:103所示的氨基酸序列或与其具有至少90％同一性的scBaff。
70.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述scTNFSF配体为具有如SEQ ID NO:104所示的氨基酸序列或与其具有至少90％同一性的scGITRL。
71.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述scTNFSF配体为具有如SEQ ID NO:105所示的氨基酸序列或与其具有至少90％同一性的sc41BBL。
72.如第1-28项和第35-58项中任一项所述的融合蛋白，其中，如ii)所述的结构域包含scTNFSF配体，并且其中所述scTNFSF配体为具有如SEQ ID NO:106所示的氨基酸序列或与其具有至少90％同一性的GITLL。
73.如前述任一项所述的融合蛋白，其中，所述细胞表面的结构或胞外基质的结构为免疫细胞的细胞表面的结构或胞外基质的结构。
74.如前述任一项所述的融合蛋白，其中，所述细胞表面的结构或胞外基质的结构为成纤维细胞的细胞表面的结构或胞外基质的结构。
75.如前述任一项所述的融合蛋白，其中，所述细胞表面的结构或胞外基质的结构为肿瘤细胞的细胞表面的结构或胞外基质的结构。
76.一种用于药物的组合物，其包含如前述任一项所述的融合蛋白。
77.一种核酸或一组核酸，其编码如第1-75项中任一项所述的融合蛋白。
78.一种生产如第1-75项中任一项所述的融合蛋白的方法，所述方法包括在至少一种类型的宿主细胞中表达如第77项所述的核酸或一组核酸，并收获所述融合蛋白。
附图说明
图1：TNFSF和TNFRSF的配体和受体。
图2：受体特异性IgG引起的TNFRSF受体的FcγR依赖的刺激。用鼠FcγR2B的表达质粒或空载体转染的Hek293细胞与所示细胞系的细胞共培养，所示细胞系中所示TNFRSF受体的刺激导致IL8的产生。在与各种TNFRSF受体特异性IgG(指示克隆名称)孵育过夜后，使用ELISA捕获IL8的产生。请注意，TNFR1和LTβR是已经被可溶性配体三聚体强烈激活的TNFRSF成员示例，并且在不表达FcγR的细胞中，它们也可以被受体特异性抗体强烈激活。所示的其他TNFRSF受体结合可溶性TNF配体没有或仅有限激活，并且在不存在FcγR结合的情况下不被抗体激活或仅低效地激活。然而，这些抗TNFRSF受体在抗体与FcγR结合后被强烈激活。
材料与方法：用编码鼠FcγR2B或空载体的表达质粒pCMV-SPORT6(SourceBicoscience)瞬时转染不表达(或仅适度表达)所示TNFRSF受体和FcγR的Hek293细胞(ATCC)。如Kums et al.,MAbs.2017Apr；9(3):506-520中所述，使用聚乙烯亚胺(PEI)将FcγR表达质粒或空载体转染Hek293细胞。第二天，收获Hek293转染子，将20.000个细胞的等分试样加入到前一天已接种(10000–20000个细胞/孔)的96孔板的孔中，这些细胞可以响应目标TNFRSF受体的激活并产生IL8。然后将共培养物用所示的抗TNFRSF受体抗体(3μg/ml)刺激过夜，一式三份，最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图3：受体特异性IgG引起的TNFRSF受体TNFR2的FcγR依赖的刺激。用所示FcγR2B的表达质粒或空载体转染的Hek293细胞与刺激TNFR2诱导产生IL8的细胞共培养。在与所示TNFR2特异性IgG孵育过夜后，使用ELISA捕获IL8的产生。
材料与方法：用编码人活性FcγR CD32A(FcγR2IIA)、人抑制性FcγR CD32B(FcγRIIB)或空载体的表达质粒pCMV-SPORT6(来自Bicoscience)瞬时转染不表达TNFR2或FcγR的Hek293细胞(ATCC)。如Kums et al.,MAbs.2017Apr；9(3):506-520中所述，使用聚乙烯亚胺(PEI)将FcγR表达质粒或空载体转染Hek293细胞。第二天，收获Hek293转染子，将20.000个细胞的等分试样加入到前一天已接种稳定表达TNFR2的HT1080细胞(密度为20000个细胞/孔)的96孔板的孔中。然后将共培养物用所示的抗人TNFR2抗体(1μg/ml)刺激过夜，一式三份，所述抗体由自身生成并生产。显示了被抗体识别的TNFR2的富含半胱氨酸的结构域(CRD)亚结构域(CRD1-CRD4)。所使用的细胞中TNFR2的激活导致增强的IL8产生。因此，为了确定TNFR2活性，最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图4：抗CD40 IgG1 G28.5的scBaff融合蛋白与BaffR、TACI或BCMA结合后，显示出增强的CD40刺激活性。(A)融合蛋白的结构。(B)空载体与分别编码BaffR、BCMA和TACI的表达质粒转染Hek293细胞。第二天，将转染子与HT1080-CD40细胞共培养，所述细胞不表达BaffR、BCMA和TACI，但在CD40刺激后强烈产生IL8。用指示浓度的抗CD40(G28.5)-HC:scBaff刺激共培养物，一式三份。次日，在不同共培养物的上清液中测定人IL8的量。(C)将表达BaffR和TACI的L363细胞、Hek293细胞(BaffR、BCMA和TACI阴性)和HT1080-CD40细胞共培养，然后用指示浓度的抗CD40(G28.5)-HC:scBaff刺激共培养物。第二天，在共培养物的上清液中再次测定人IL8的量。请注意，scBaff中Baff的三个原聚体仅包含全长Baff分子的胞外TNF同源结构域。
材料与方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的Hek293细胞和FcgR表达载体，用编码BaffR、TACI和BCMA的表达质粒或空载体(＝阴性对照)瞬时转染不表达CD40、BaffR、TACI或BCMA的Hek293细胞。收获转染子，并将30.000个细胞的等分试样加入到前一天已以20000个细胞/孔的密度接种了HT1080-CD40细胞(稳定转染CD40的HT1080细胞)的96孔板的孔中。可选地，将HT1080-CD40细胞与50.000个细胞/孔的内源性表达BaffR和TACI的骨髓瘤细胞系L363共培养。所述HT1080-CD40细胞对CD40的激活产生大量响应。将共培养物用TRAAFFIAA抗CD40(G28.5)-IgG1(N297)-scBaff刺激过夜，一式三份，最后通过ELISA测定IL8的产量。
图5：抗CD95 IgG1 E09的scBaff融合蛋白与BaffR、TACI或BCMA结合后显示出增加的CD95刺激活性。(A)融合蛋白的结构。(B)空载体或编码BaffR、BCMA和TACI的表达质粒转染Hek293细胞。第二天，将转染子与HT1080细胞共培养，所述细胞不表达BaffR、BCMA和TACI，并在CD95刺激后发生凋亡。用指示浓度的抗CD95(E09)-HC:scBaff处理共培养物并在次日通过结晶紫染色评估粘附生长的HT1080细胞的活性。(C)将表达BaffR和TACI的L363细胞、Hek293细胞(BaffR、BCMA和TACI阴性)与HT1080细胞共培养。再次用抗CD95(E09)-HC:scBaff刺激共培养物过夜，最后通过结晶紫染色评估HT1080的活性。请注意，scBaff中Baff的三个原聚体仅包含全长Baff分子的胞外TNF同源结构域。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的Hek293细胞和FcgR表达载体，用编码BaffR、TACI和BCMA的表达质粒或空载体(＝阴性对照)瞬时转染不表达BaffR、TACI或BCMA和表达可忽略量的CD95的Hek293细胞。收获转染子，并将30.000个细胞的等分试样加入前一天已以20000细胞/孔的密度接种HT1080细胞的96孔板的孔中。可选地，将HT1080细胞与50.000细胞/孔的内源性表达BaffR和TACI的骨髓瘤细胞系L363共培养。在没有敏化剂的情况下，HT1080细胞对CD95诱导的细胞死亡敏感。将共培养物用TRAAFFIAA抗CD95(E09)-IgG1(N297)-scBaff刺激过夜，一式三份，最后通过粘附生长的HT1080细胞的结晶紫染色测定细胞死亡诱导。
图6：结合TNFR2的抗CD40-IgG1融合蛋白在结合TNFR2后具有增强的CD40刺激活性。将既不表达TNFR2也不表达CD40的HeLa细胞、在TNFR2刺激后仅产生中等水平IL8的HeLa-TNFR2转染子与既不表达TNFR2也不表达CD40的HT1080细胞以及在CD40刺激后产生非常高水平的IL8的HT1080-CD40转染子共培养，并用指示浓度的靶向TNFR2的抗CD40融合蛋白(抗CD40(G28.5)-HC:scTNF80)刺激。第二天，测定不同共培养物的上清液中的IL8的量。请注意，scTNF80中TNF80的三个原聚体仅包含全长鼠TNF分子的胞外TNF同源结构域，包含赋予TNFR2特异性的突变。如前所述，以96孔形式进行测定，一式三份。
图7：结合CD19的抗CD95-IgG1融合蛋白与表达CD19的细胞共培养时，在HT1080细胞中显示增强的细胞死亡诱导。将不表达CD19的CD95敏感细胞系HT1080与Jurkat(CD19阴性)细胞和BJAB(CD19阳性)细胞共培养，并与结合CD19的抗CD95融合蛋白(抗CD95(E09)-HC:scFv:CD19)以指示浓度孵育。第二天，通过结晶紫染色测定粘附生长的HT1080细胞的活性。
材料与方法：实验在96孔板上进行，一式三份。HT1080细胞按每孔20000个细胞接种。接下来加入Jurkat和BJAB细胞(每孔30000个)以及指示浓度的抗CD95(E09)-HC:scFv:CD19。通过结晶紫染色贴壁生长的HT1080细胞测定细胞死亡诱导。借助于未处理的HT1080细胞(＝100％活性)和用含有CHX、Velcade和Fc-CD95L的高毒性混合物处理的HT1080细胞(＝0活性)对细胞活性归一化。
图8：TNFR2特异性抗体C4的IL2融合蛋白以依赖IL2R的方式增强刺激TNFR2。将既不表达TNFR2也不表达IL2R的Hek293细胞用编码IL2R的亚基的表达质粒的混合物或者空载体转染。然后将Hek293转染子与表达TNFR2的HeLa转染子共培养，并用指示浓度的抗TNFR2(C4)-HC:IL2刺激。第二天，使用ELISA捕获可通过TNFR2诱导产生的IL8。
材料与方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码IL-2受体亚基的表达质粒混合物瞬时转染Hek293细胞，或空载体作为阴性对照瞬时转染Hek293细胞。收获转染子，将20.000个细胞的等分试样加入前一天已以20000个细胞/孔的密度接种了HeLa-TNFR2细胞的96孔板的孔中。所述HeLa-TNFR2细胞响应TNFR2的激活而产生IL8。将共培养物用TRAAFFIAA抗TNFR2(C4)-HC:IL2刺激过夜，一式三份，最后通过ELISA测定IL8的产生。
图9：TNFR2特异性抗体C4的锚定GITR和41BB的融合蛋白在结合GITR和41BB后显示出增强的TNFR2活化。(A)所使用的刺激TNFR2的锚定人GITR和鼠4-1BB的TRAAFFIAA的域结构。(B，C)用空载体或编码人GITR(B)或鼠41BB(C)的表达质粒转染既不表达GITR也不表达41BB的细胞。然后将转染子与响应TNFR2激活而产生IL8的HeLa-TNFR2细胞共培养。将共培养物用指示浓度的抗TNFR2(C4)-HC:scGITRL(B)和抗TNFR2(C4)-HC:sc(mu)4-1BBL(C)刺激。第二天，采用IL8特异性ELISA测定IL8的产生。(D)所使用的鼠GITR锚定的TNFR2刺激的TRAAFFIAA的域结构。(E)将用空载体或鼠GITR的表达质粒转染的细胞与HeLa-TNFR2细胞共培养。在用抗TNFR2(C4)-HC:(mu)GITRL刺激后，通过ELISA监测第二天IL8的产生。请注意，与人GITRL以及TNFSF的所有其他人和鼠配体相反，鼠GITRL是二聚体分子，并与两个而不是三个受体分子相互作用。因此，将鼠GITRL的单个原聚体融合到重链的C末端，以构建单个(mu)GITRL二聚体，其作为相应抗体-(mu)GITRL的锚定结构域。相反，使用scTNFSF配体(例如scGITRL、sc(mu)41BBL和scTNF80)作为锚定结构域融合至重链，使得每个抗体融合蛋白产生两个锚定结构域。请注意，scGITRL和sc(mu)41BBL中的GITRL和(mu)41BBL的三个原聚体仅包含全长配体分子的胞外TNF同源结构域。同样，(mu)GITRL仅指鼠全长鼠GITRL的THD。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码人或鼠GITR或鼠41BB的表达质粒或空载体(＝阴性对照)瞬时转染不表达GITR、41BB或TNFR2的Hek293细胞。收获转染子，将20000个细胞的等分试样加入已以20000个细胞/孔的密度接种HeLa-TNFR2细胞的96孔板的孔中。所述HeLa-TNFR2细胞响应TNFR2激活而产生IL8。将共培养物用所示的TRAAFFIAA刺激过夜，一式三份，最后通过ELISA测定IL8的产生。
图10：锚定细胞表面抗原CD20的CD40特异性抗体G28.5的scFv融合蛋白以依赖CD20的方式增强刺激CD40。将CD20阴性Jurkat细胞和CD20阳性BJAB细胞与可应答CD40的HT1080-CD40细胞共培养，并用指示浓度的抗CD40(G28.5)-HC:scFv:CD20刺激。第二天，使用ELISA捕获可通过CD40诱导产生的IL8。使用具有N297A突变的IgG1分子的变体，所述变体在体内具有抑制Fc结构域的ADCC活性。
材料与方法：实验在96孔板上进行，一式三份。以每孔20000个细胞接种HT1080-CD40细胞。第二天加入Jurkat(CD20-)和BJAB细胞(CD20+)(每孔30000个)，以及指定浓度的抗CD40-HC:scFv:CD20。最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图11：抗体E09的抗CD95-IgG1 Fab2片段的scFv:CD20融合蛋白在与表达CD20的细胞共培养时，在HT1080细胞中显示出增强的细胞死亡诱导。将CD95敏感细胞系HT1080与Jurkat(CD20阴性)细胞和BJAB细胞(CD20阳性)共培养。用指示浓度的结合CD20的抗CD95融合蛋白抗CD95-Fab2-HC:scFv:CD20处理共培养物。第二天，通过结晶紫染色确定粘附生长的HT1080细胞的生存力。
材料与方法：实验在96孔板上进行，一式三份。以每孔接种20000个细胞接种HT1080细胞。接下来共同添加Jurkat和BJAB细胞(每孔30000个)以及指示浓度的TRAAFFIAA。借助于未处理的HT1080细胞(＝100％活性)和用含有CHX、Velcade和Fc-CD95L(＝0活性)的高毒性混合物处理的HT1080细胞对HT1080的细胞活性归一化。
图12：TNFR2特异性抗体C4的靶向细胞表面抗原的scFv融合蛋白以依赖抗原的方式增强刺激TNFR2。将CD19和CD20阴性的Jurkat细胞以及CD19和CD20阳性的Raji细胞与表达TNFR2的HeLa转染子共培养，并分别用指示浓度的抗TNFR2-IgG1(N297A)-HC:scFv:CD19(上图)或抗TNFR2-IgG1(N297A)-HC:scFv:CD20(下图)刺激。第二天，使用ELISA捕获可通过TNFR2诱导的IL8的产生。使用了不能触发ADCC的IgG1变体N297A。这里使用的亲本TNFR2特异性mAb C4是内部制备的。
材料与方法：实验在96孔板上进行，一式三份。上图：以每孔20000个细胞接种HeLa-TNFR2细胞。第二天，加入Jurkat(CD19-/CD20-)和RAJI细胞(CD19-/CD20-)(每孔30000)以及指示浓度的抗TNFR2-IgG1(N297A)-HC:scFv:CD19(上图)或者抗TNFR2-IgG1(N297A)-HC:scFv:CD20(下图)。最后通过ELISA(BD Biosciences)分析评估共培养上清液中由TNFR2激活产生的IL8含量。
图13：抗CD40 G28.5的scBaff融合蛋白的生化性质。(A)纯化的抗CD40(G28.5)-IgG1(N297)(I)、抗CD40(G28.5)-IgG1(N297)-HC:scBaff(II)、抗CD40(G28.5)-FAB2(III)和抗CD40(G28.5)-FAB2-HC:scBaff(IV)通过SDS-PAGE分离。然后通过可识别各种抗体融合蛋白的重链和轻链中存在的Flag表位的mAb M2进行Western blotting(左图)或银染(右图)对蛋白进行可视化。请注意，构建体II的两条链的大小非常相似，因此无法通过SDS-PAGE分离。(B)标记蛋白混合物和蛋白I至IV的凝胶过滤分析。(C)蛋白质I至IV的示意图。
材料与方法：抗体融合蛋白的制备和纯化如Kums et al.,SDS-PAGE,2017所述，银染和western blotting的方法如Lang et al.,2016所述。
Kums J,Nelke J,RüthB,
Lang I,Füllsack S,Wyzgol A,Fick A,Trebing J,Arana JA,Weisenberger D,Wajant H,Binding Studies of TNF Receptor Superfamily(TNFRSF)Receptors on Intact Cells.J Bio Chem.2016Mar 4；291(10):5022-37。
图14：抗CD40 G28.5的IgG1(N297A)与FAB2变体的scBaff融合蛋白在与内源性表达BaffR和TACI的淋巴瘤/骨髓瘤细胞结合后，表现出增强的CD40刺激活性。将MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞或缺少所有Baff结合受体表达的Jurkat细胞作为对照，与不表达BaffR、BCMA和TACI但在CD40刺激后会强烈产生IL8的HT1080-CD40细胞共培养。用指示浓度的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff或抗CD40(G28.5)-FAB2-HC:scBaff刺激共培养物，一式三份。第二天，在不同共培养物的上清液中测定人IL8的量。
材料与方法：HT1080-CD40细胞在96孔板中培养过夜(每孔20.000个细胞)。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat细胞，并以指示浓度的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff刺激所得共培养物过夜，一式三份。然后通过ELISA分析细胞上清液中产生的IL8。请注意，与HT1080-CD40细胞相比，MM1S、L363、BJAB和Jurkat细胞不产生IL8或产生可忽略不计的IL8。
图15：与可溶性Baff(TNC-Baff)的竞争抑制了抗CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff中IL8的产生。HT1080-CD40细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞共培养，Jurkat细胞作为对照。请注意，HT1080-CD40和Jurkat细胞不表达Baff相互作用受体(BaffR、TACI、BCMA)。在存在和不存在过量可溶性TNC-Baff(5μg/ml)的情况下，用200ng/ml的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff或抗CD40(G28.5)-FAB2-HC:scBaff刺激共培养，一式三份。第二天，在各种共培养物的上清液中测定人IL8的量。
材料与方法：HT1080-CD40细胞在96孔板中培养过夜(每孔20.000个细胞)。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat细胞，并在存在和不存在可溶性TNC-Baff(5μg/ml)的情况下，用200ng/ml CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff刺激所得共培养物过夜，一式三份，所述可溶性TNC-Baff是含有Tenascin-C三聚结构域的可溶性Baff的稳定形态(Berg et al.,2007)。然后通过ELISA分析细胞上清液中IL8的产生。请注意，与HT10890-CD40细胞相比，MM1S、L363、BJAB和Jurkat细胞不产生IL8或产生可忽略不计的IL8。
Berg D,Lehne M,MüllerN,Siegmund D,Münkel S,Sebald W,Pfizenmaier K,Wajant H,Enforced covalent trimerization increases the activity of the TNFligand family members TRAIL and CD95L,Cell Death Differ.2007Dec；14(12):2021-34,EPUB 2007Aug 17。
图16：抗CD95 IgG1 E09或其FAB2片段的scBaff融合蛋白与具有BaffR和TACI内源性表达的淋巴瘤/骨髓瘤悬浮细胞结合后，诱导贴壁HT1080细胞死亡。将贴壁生长的HT1080细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)悬浮细胞共培养，Jurkat细胞作为对照。请注意，HT1080和Jurkat细胞不表达与Baff相互作用的受体(BaffR、TACI、BCMA)。在1μg/ml CHX的存在下，用指示浓度的抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激共培养物，一式三份。第二天，通过剩余附着的HT1080细胞的结晶紫染色确定细胞活力。
材料与方法：HT1080细胞的接种密度为20.000个细胞/孔。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat悬浮细胞，并将所得的共培养物用1μg/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激过夜，一式三份。最后，通过贴壁生长的HT1080细胞的结晶紫染色确定细胞死亡诱导。借助未经处理的HT1080细胞(＝100％活力)和以0.03％叠氮、50μg/ml CHX和500ng/ml Fc-CD95L(＝0％活力)的高毒性混合物处理的HT1080细胞的活力进行了归一化。
图17：与可溶性Baff(TNC-Baff)的竞争抑制了HT1080细胞和Baff相互作用受体表达细胞的共培养物中抗CD95(E09)-IgG1(N297A)-HC:scBaff和抗CD95(E09)-FAB2-HC:scBaff诱导的HT1080杀伤。HT1080细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞共培养，Jurkat细胞作为对照。请注意，HT1080-CD40和Jurkat细胞不表达Baff相互作用受体(BaffR、TACI、BCMA)。在存在和不存在过量的可溶性TNC-Baff(5μg/ml)的情况下，以1μg/ml CHX刺激共培养物与20ng/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff，一式三份。第二天，通过剩余附着的HT1080细胞的结晶紫染色确定细胞活力。
材料与方法：HT1080细胞的接种密度为20.000个细胞/孔。第二天，每孔加入w0.000个MM1S、L363、BJAB或Jurkat悬浮细胞，并在存在和不存在可溶性TNC-Baff(5μg/ml)的情况下，用20ng/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激过夜共培养物，一式三份。最后，通过贴壁生长的HT1080细胞的结晶紫染色确定细胞死亡诱导。借助未经处理的HT1080细胞(＝00％活力)和以0.03％叠氮、50μg/ml CHX和500ng/ml Fc-CD95L(＝0％活力)的高毒性混合物处理的HT1080细胞的活力进行了归一化。TNC-Baff是包含tenascin-C的三聚结构域的可溶性Baff的一种稳定形态(Berg et al.,2007)。
Berg D,Lehne M,Müller N,Siegmund D,Münkel S,Sebald W,Pfizenmaier K,Wajant H.,Enforced covalent trimerization increases the activity of the TNFligand family members TRAIL and CD95L,Cell Death Differ,2007Dec；14(12):2021-34。
图18：抗CD95(E09)-IgG1(N297A)-HC:scBaff而不是抗CD95(E09)-IgG1(N297A)激活BaffR表达细胞中的凋亡半胱氨酸蛋白酶。将BJAB细胞(表达BaffR)和作为阴性对照的不表达任何Baff受体(BaffR、TACI、BCMA)的Jurkat细胞用指示浓度的抗CD95(E09)-IgG1(N297A)-HC:scBaff和抗CD95(E09)-IgG1(N297A)处理过夜。第二天，通过westernblotting分析总细胞裂解物的caspase-8的p18片段和caspase-3的p17片段的生成，两种片段是这些半胱氨酸蛋白酶的凋亡活化的指示剂。作为阳性对照，用500ng/ml Fc-CD95L处理Jurkat和BJAB细胞，所述两种细胞系均可被其诱导凋亡。
图19：IgG2同种型的TNFR2特异性抗体C4锚定细胞表面抗原CD20的scFv融合蛋白以依赖CD20的方式增强刺激TNFR2。将CD20阴性Jurkat和CD20阳性BJAB细胞与TNFR2反应性HeLa-TNFR2细胞共培养，并用指示浓度的抗TNFR2(C4)-IgG2-HC:scFv:CD20刺激。第二天，使用ELISA测定了可以经由TNFR2诱导产生的IL8。
材料与方法：以每孔20.000个HeLa-TNFR2转染子(Weiss et al.,1997)接种在96孔板中。第二天，加入CD20阳性BJAB细胞或作为阴性对照的20.000个缺乏内源性表达CD20的Jurkat细胞。然后用指示浓度的抗TNFR2(C4)-IgG2-HC:scFvCD20处理共培养物，次日通过ELISA测定IL8的产生。
Weiss T,Grell M,Hessabi B,Bourteele S,Müller G,Scheurich P,WajantH.v,Enhancement of TNF receptor p6-mediated cytotoxicity by TNF receptor p80:requirement of the TNF receptor-associated factor-2binding site,J Immunol,1997Mar 1；158(5):2398-404。
图20：IgG1同种型的TNFR2特异性抗体C4锚定细胞表面抗原CD70的scFv融合蛋白以依赖CD27L/CD70r方式增强刺激TNFR2。(A)融合蛋白的结构。(B，C)用空载体或编码CD27L(或称CD70)的表达质粒转染Hek293细胞。第二天，将转染子与在TNFR2表达时产生IL8的HeLa-TNFR2转染子共培养。用指示浓度的抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(1F6)(B)或抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(2H5)处理共培养物(C)。最后，通过ELISA测定IL8的产生。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码CD27L的表达质粒(＝阴性对照)瞬时转染不表达TNFR2或CD27L(CD70)的Hek293细胞。收获转染子，将20.000个细胞等分添加到前一天已以20.000个细胞/孔的密度接种了HeLa-TNFR2细胞的96孔板的孔中。用TRAAFFIAA，即抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(1F6)(B)和抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(2H5)(C)刺激共培养物过夜，一式三份。2H5和1F6是两种不同的人CD70特异性抗体。
图21：4-1BB特异性抗体HBBK4、CD40特异性抗体G28.5和CD95特异性抗体E09的IgG1(N297)变体的scFv:CD20融合蛋白诱导与表达CD20的细胞共培养的HT1080-41BB、HT1080-CD40和HT1080(内源性表达CD95)细胞中IL8的产生。(A)融合蛋白示意图。(B-D)将所示的HT1080变体与用空载体或CD20表达质粒转染的Hek293细胞共培养。然后用指示浓度的结合CD20的IgG1(N297A)融合蛋白抗41BB(HBBK4)-IgG1(N297A)-HC:scFv:CD20(B)、抗CD40(B28.5)-IgG1(N297A)-HC:scFv:CD20(C)和抗CD95(E09)-IgG1(N297A)-HC:scFv:CD20(D)处理共培养物。第二天，通过ELISA测定IL8的产生。对于抗CD95变体，加入半胱氨酸蛋白酶抑制剂ZVAD以防止细胞凋亡。
材料与方法：HT1080-41BB转染子(Wyzgol et al.,2009)、HT1080-CD40转染子(Wyzgol et al.,2009)和HT1080细胞(内源性表达CD95)以20.000个细胞/孔的密度接种。第二天，加入20.000个已于前一天用空载体(EV)或CD27L(＝CD70)表达质粒转染的Hek293细胞。用指示浓度的抗41BB(HBBK4)-IgG1(N297A)-HC:scFvCD20(B)、抗CD40(G28.5)-IgG1(N297A)-HC:scFvCD20(C)和抗CD95(E09)-IgG1(N297A)-HC:scFvCD20(D)刺激共培养物过夜，一式三份。后者在20μg/ml ZVAD的存在下加入，以防止细胞凋亡诱导。通过ELISA测定IL8。
Wyzgol A,Müller N,Fick A,Munkel S,Grigoleit GU,Pfizenmaier K,WajantH.,Trimer stabilization,oligomerization,and antibody-mediated cell surfaceimmobilization improve the activity of soluble trimers of CD27L,CD40L,41BBL,and glucocorticoid-induced TNF receptor ligand,J Immunol,2009Aug 1；183(3):1851-61。
图22：4-1BB特异性抗体HBBK4、CD40特异性抗体G28.5和CD95特异性抗体E09的IgG1(N297)变体的scFv:CD20融合蛋白诱导与CD20阳性的BJAB细胞共培养的HT1080-41BB、HT1080-CD40和HT1080(内源性表达CD95)细胞中增强的IL8产生。(A-C)将所示的HT1080变体与CD20阳性BJAB细胞共培养，缺少内源表达CD20的Jurkat细胞作为阴性对照。然后用指示浓度的结合CD20的IgG1(N297A)融合蛋白抗41BB(HBBK4)-IgG1(N297A)-HC:scFv:CD20(A)、抗CD40(G28.5)-IgG1(N297A)-HC:scFv:CD20(B)和抗CD95(E09)-IgG1(N297A)-HC:scFv:CD20(C)处理共培养物。第二天，通过ELISA测定IL8的产生。对于抗CD95变体，加入半胱胺酸蛋白酶抑制剂ZVAD以防止细胞凋亡。
材料与方法：HT1080-41BB转染子(Wyzgol et al.,2009)、HT1080-CD40转染子(Wyzgol et al.,2009)和HT1080细胞(内源性表达CD95)以20.000个细胞/孔的密度接种。第二天，加入BJAB细胞(CD20阳性)或20.000个Jurkat细胞(CD20阴性)。用指示浓度的抗41BB(HBBK4)-IgG1(N297A)-HC:scFvCD20(A)、抗CD40(G28.5)-IgG1(N297A)-HC:scFvCD20(B)和抗CD95(E09)-IgG1(N297A)-HC:scFvCD20(C)刺激共培养物过夜，一式三份。后者在20μg/ml ZVAD的存在下加入，以防止细胞凋亡诱导。通过ELISA测定IL8。
Wyzgol A,Müller N,Fick A,Munkel S,Grigoleit GU,Pfizenmaier K,WajantH.,Trimer stabilization,oligomerization,and antibody-mediated cell surfaceimmobilization improve the activity of soluble trimers of CD27L,CD40L,41BBL,and glucocorticoid-induced TNF receptor ligand,J Immunol,2009Aug 1；183(3):1851-61。
图23：TRAILR2特异性抗体Conatumumab(Cona)的IgG1(N297A)同种型与CD70特异性scFv 9G2的融合蛋白以依赖CD70的方式增强刺激促炎性(A)和细胞毒性(B)TRAILR2信号转导。用编码CD70的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与TRAILR2反应性HT1080细胞(无CD70表达)共培养。为使HT1080细胞对TRAILR2信号敏感，用2.5μg/ml CHX(B)或CHX加20μM半胱氨酸抑制剂ZVAD(A)处理共培养物。CHX抑制具有高周转率的TRAILR2信号抑制剂FLIP蛋白的表达。ZVAD可以防止细胞凋亡，从而可以评价TRAILR2的基因诱导特性。然后用指示浓度的抗TRAILR2(Cona)-IgG1(N297A)-HC:scFv:CD70(9G2)进一步处理共培养物，再过一天后，通过ELISA(A)测定IL8的产生或通过结晶紫染色(B)测定细胞活力。
图24：IgG1(N297A)或Fab2形式的CD40特异性抗体C与衍生自靶向PD-L1的检查点抑制剂Avelumab(Ave)的scFv的融合蛋白以PD-1L限制性方式激活CD40。将编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与CD40反应性HT1080-CD40细胞共培养。然后用指示浓度的抗CD40(C)-IgG1(N297A)-HC:scFv:PD-L1(A)或抗CD40(C)-Fab2-HC:scFv:PD-L1(B)处理共培养物。第二天，通过ELISA测定IL8的产生，作为CD40激活的读数。
图25：IgG1(N297A)或Fab2形式的41BB特异性抗体HBBK与衍生自检查点抑制剂Avelumab(Ave)的scFv的融合蛋白以PD-1L限制性方式激活41BB。用编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与41BB反应性HT1080-41BB细胞共培养。然后用指示浓度的抗41BB(HBBK)-IgG1(N297A)-HC:scFv:PD-L1(A)或抗41BB(HBBK)-Fab2-HC:scFv:PD-L1(B)处理共培养物。第二天，通过ELISA测定IL41的产生，作为41BB激活的读数。
图26：PD-1L特异性抗体Avelumab(Ave)的IgG2与41BB特异性scFv的融合蛋白显示以PD-1L限制性方式增强41BB的活化。用编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与41BB反应性HT1080-41BB细胞共培养。然后用指示浓度的抗PD-L1(Ave)-IgG2-HC:scFv:41BB(HBBK)处理共培养物。第二天，通过ELISA测定IL41的产生，作为41BB激活的读数。所述实施例说明，抗TNFRSF受体抗体衍生的scFv在与识别细胞表面暴露的抗原的抗体融合后获得依赖锚定的激动剂活性。因此，TRAAFFIAA原理上与抗TNFRSF受体抗体(作为TNFRSF受体的刺激效应子结构域)和scFv(作为锚定结构域)一起作用，也以镜像形式与抗体(作为锚定结构域)和TNFRSF受体特异性scFv(作为效应子结构域)起作用。
图27：鼠CD27特异性抗体的IgG(N297A)与Fn14特异性scFv 18D1的融合蛋白显示出以鼠CD27限制性方式增强的Fn14活化。将编码鼠CD27的表达质粒瞬时转染的Hek293细胞(对Fn14响应差)或空载体瞬时转染的Hek293细胞与高度Fn14响应的HT1080细胞共培养。然后用指示浓度的抗muCD27-IgG1(N297A)-HC:scFv14(18D1)处理共培养物。第二天，通过ELISA测定IL8的产生，作为Fn14激活的读数。所述实施例再次说明，抗TNFRSF受体抗体衍生的scFv在与识别细胞表面暴露的抗原的抗体融合后获得依赖锚定的激动剂活性。因此，TRAAFFIAA原理上与抗TNFRSF受体抗体(作为TNFRSF受体的刺激效应子结构域)和scFv(作为锚定结构域)一起作用，也以镜像形式与抗体(作为锚定结构域)和TNFRSF受体特异性scFv(作为效应子结构域)起作用。
发明详述
定义和通用技术
除非下文另外定义，否则应根据本领域技术人员已知的通用含义来理解本发明中使用的术语。
本文引用的所有出版物、专利和专利申请出于所有目的通过引用整体并入本文。
本文所使用的“scTNFSF”是指单链形式的功能性(意味着受体结合)TNFSF配体。通常，一个scTNFSF包含三个单元的TNFSF原聚体的受体结合结构域，其中所述单元通过多肽接头连接并形成分子内三聚体。
本文所使用的术语“抗体”是指能特异性结合目的抗原的任何功能性抗体，如Paul,W.E,(Ed.).:Fundamental Immunology 2nd Ed.,Raven Press,Ltd.,New York,1989的第7章中概述，其通过引用并入本文。在没有特别限制的情况下，术语“抗体”包括来自任何适当来源物种的抗体，包括鸡和哺乳动物，例如鼠、山羊、非人灵长类动物和人。优选地，所述抗体是人源化抗体。所述抗体优选为可以通过本领域公知的方法制备的单克隆抗体。术语“抗体”包括IgG-1、-2、-3或-4、IgE、IgA、IgM或IgD同种型抗体。术语“抗体”包括单体抗体(例如IgD、IgE、IgG)或寡聚抗体(例如IgA或IgM)。术语“抗体”还包括但不限于分离的抗体和修饰的抗体，比如基因工程抗体，例如嵌合或人源化抗体。
所述抗体结构域的命名遵循本领域已知的术语。如本领域公知的，抗体的每个单体包含两个重链和两个轻链。其中，每条重链和轻链均包含可变结构域(对于重链而言称为VH，对于轻链而言称为VL)，这对于抗原结合而言是重要的。这些重链和轻链可变结构域包含(以N末端至C末端的顺序)区域FR1、CDR1、FR2、CDR2、FR3、CDR3和FR4(FR，构架区；CDR，互补决定区，也称为高变区)。抗体序列内上述抗体区域的鉴定和分配通常根据Kabat et al.(Sequences of proteins of immunological interest,U.S.Dept.of Health and HumanServices,Public Health Service,National Institutes of Health,Bethesda,Md.1983)或Chothia et al.(Conformations of immunoglobulin hypervariableregions,Nature,1989 Dec 21-28；342(6252):877-83.)的方法，或者可以通过使用在Giudicelli et al.(IMGT/V-QUEST,an integrated software program forimmunoglobulin and T cell receptor V-J and V-D-J rearrangement analysis,Nucleic Acids Res.2004 Jul 1；32(Web Server issue):W435-40)-在此通过引用并入本文-中所述的IMGT/V-QUEST软件来进行。优选地，通过使用IMGT/V-QUEST软件鉴定和分配上述抗体区域。
“单克隆抗体”是来自基本上同质的抗体群的抗体，其中所述抗体在序列上基本上相同(即，除了含有天然存在的序列修饰例如在其N-和C-末端的氨基酸修饰的抗体的小部分不同以外)。与包含针对单个表位或针对许多不同表位的不同抗体的混合物的多克隆抗体不同，单克隆抗体针对同一表位，因此具有高度特异性。术语“单克隆抗体”包括(但不限于)从衍生自单细胞克隆的单克隆细胞群体获得的抗体，例如通过
“人源化抗体”是包含人序列和赋予与目标抗原结合特异性的一小部分非人序列的抗体。通常，人源化抗体是通过用与目标抗原结合的非人供体抗体(例如小鼠、兔、大鼠供体抗体)的高变区序列替换人受体抗体的高变区序列而产生的。在一些情况下，受体抗体的构架区序列也可以被供体抗体的相应序列替换。除了衍生自供体和受体抗体的序列外，“人源化抗体”可以包含其他(附加的或替代的)残基或序列，也可以不包含。此类其他残基或序列可用于进一步改善抗体特性，例如结合特性(例如降低Kd值)和/或免疫原性特性(例如降低人的抗原性)。产生人源化抗体的方法的非限制性实例在本领域是已知的，例如来自Riechmann et al.(Nature,1988 Mar 24；332(6162):323-7)或Jones et al.(Nature,1986 May 29-Jn 4；321(6069):522-5)。
术语“人抗体”是指包含人可变和恒定结构域序列的抗体。该定义涵盖具有单个氨基酸取代或修饰的人序列的抗体，其可用于进一步改善抗体特性，例如结合特性(例如降低Kd值)和/或免疫原性特性(例如降低人类的抗原性)。术语“人抗体”不包括赋予与目的抗原结合特异性的部分非人序列的人源化抗体。
本文所使用的抗体的“抗原结合部分”是指保留抗体特异性结合抗原的能力的抗体部分。所述能力可以，例如通过本领域已知的方法，通过确定抗原结合部分与抗体抗原特异性结合的能力来确定。抗原结合部分可以包含抗体的一个或多个片段。没有特别限制，所述抗原结合部分可以通过本领域已知的任何合适方法来产生，包括重组DNA方法和通过抗体的化学或酶促片段制备。抗原结合部分可以是Fab片段、F(ab')片段、Fab2片段、单链抗体(scFv)、单结构域抗体、双抗体或抗体的其他任何保留抗体抗感染能力的部分与抗原特异性结合。应当理解，根据本领域中已知的术语“Fab2”的含义，术语“Fab2”与术语“Fab2”、“Fab2”和“FAB2”同义。因此，出于本申请的目的，术语“Fab2”与术语“Fab2”、“Fab2”和“FAB2”可互换使用。根据本发明的“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”没有特别限制。例如，它可以是能结合肿瘤抗原的抗原结合部分。所述“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”优选选自抗CD20抗原结合部分、抗CD70抗原结合部分、抗CD19抗原结合部分、抗EGFR抗原结合部分、抗Her2抗原结合部分、抗Fn14抗原结合部分、抗CD40L抗原结合部分和抗PD1L抗原结合部分。可选地，根据本发明的“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”可以是抗FAP抗原结合部分、抗BCMA抗原结合部分或者抗Flt3抗原结合部分。
“抗体”(例如单克隆抗体)或者“抗原结合部分”可能已被衍生化或连接至不同的分子。例如，可以与抗体连接的分子是其他蛋白质(例如其他抗体)、分子标记(例如荧光、发光、有色或放射性分子)、药剂。所述抗体或抗原结合部分可以直接连接(例如以两种蛋白质之间的融合形式)，或者通过接头分子(例如本领域已知的任何合适类型的化学接头)连接。
有关包括本发明所述融合蛋白的本发明，“抗TNFRSF受体抗体或其抗原结合部分”的含义没有特别限制，只要它是针对至少一种TNFRSF受体的抗体或其抗原结合部分即可。在根据本发明的所有其他实施方案的优选实施方案中，所述抗TNFRSF受体抗体或其抗原结合部分可以是针对单一类型的TNFRSF受体的抗TNFRSF受体抗体或其抗原结合部分。在根据本发明所有其他实施方案的另一个优选实施方案中，所述抗TNFRSF受体抗体或其抗原结合部分可以是针对两种不同类型的TNFRSF受体的二价抗TNFRSF受体抗体或其抗原结合部分。
本文所使用的术语“结合”是指与目的抗原的特异性结合。优选地，所述Kd值小于100nM，更优选地小于50nM，进一步优选地小于10nM，更进一步优选地小于5nM，最优选地小于2nM。
本文所使用的术语“表位”是指抗原的一小部分，其形成抗体的结合位点。
在本发明的上下文中，出于表征抗体或抗体融合蛋白的结合特性的目的，本发明的抗体或者所述融合蛋白(例如TRAAFFIAA)任何结合或者竞争性结合目的抗原或者FcγR或者所述细胞表面或所述胞外基质的所述结构，优选使用荧光素酶标记的(例如Gaussiaprinceps luciferase(GpL))抗体或融合蛋白的变体，通过细胞结合研究(例如Kums etal.,MAbs,2017Apr；9(3):506-520中所述)或者表面等振离子共振测量作为参考标准测定。
术语“KD”或“KD值”是指本领域已知的平衡解离常数。在本发明的上下文中，这些术语是指本发明的抗体或融合蛋白(例如TRAAFFIAA)相对于具体的目的抗原或者目的FcγR或者相对于细胞表面或胞外基质的所述结构的平衡解离常数。所述平衡解离常数是复合物(例如抗原-抗体复合物)可逆地解离为其组分(例如抗原和抗体)的倾向的量度。对于根据本发明的抗体或融合蛋白，KD值优选通过GpL标记的所述抗体或融合蛋白(例如TRAAFFIAA)的变体与未修饰的目的抗体或融合蛋白(例如TRAAFFIAA)的胞内竞争性结合研究或表面等振离子共振测定。
本文所使用的结合本发明的所述抗体或其抗原结合部分，术语“能与抗体交叉竞争”以与具体蛋白质例如TNFRSF受体结合通常是指在使用本领域已知的分析方法，例如酶联免疫吸附测定法(ELISA)时，抗体或其抗原结合部分能交叉竞争。如本领域已知的，应当理解的是，术语“能与抗体交叉竞争”是指与所述具体蛋白质特异性结合的交叉竞争。本领域技术人员将能确定用于检测这种特异性结合的交叉竞争的合适条件。
本文所使用的“分离的抗体”或“分离的TRAAFFIA”或“分离的融合蛋白”已被鉴定并与其来源环境的大部分组分(按重量计)分离，例如，来自杂交瘤细胞培养物或用于生产的不同细胞培养物的组分(例如重组表达抗体或融合蛋白(例如TRAAFFIA)的生产细胞，例如CHO或HEK293细胞)。进行所述分离是为了充分去除可能干扰所述抗体或融合蛋白(如TRAAFFIA)适用所需用途的组分(例如，根据本发明的所述抗体或融合蛋白(如TRAAFFIA)的治疗用途)。制备分离的抗体或抗体融合蛋白的方法是本领域已知的，包括蛋白A色谱法、阴离子交换色谱法、阳离子交换色谱法、病毒保留过滤和超滤。优选地，通过Lowry蛋白测定法测定，所述分离的抗体或融合蛋白(如TRAAFFIA)制剂至少为70％纯(w/w)，更优选地至少为80％纯(w/w)，进一步优选地为至少90％纯(w/w)，更进一步优选地至少95％纯(w/w)，最优选地为至少99％纯(w/w)。
本文所使用的“双抗体”是二价抗原结合抗体的一小部分，其包含重链可变结构域，所述重链可变结构域通过肽接头连接至同一多肽链上的轻链可变结构域，所述肽接头太短而无法使同一条链上的两个结构域配对。这导致与另一条链的互补结构域配对，并导致具有两个抗原结合位点的二聚体分子组装。双价抗体可以是双价和单特异性的(例如具有两个同一抗原的结合位点的双抗体)，也可以是双价和双特异性的(例如具有两个抗原结合位点的双价抗体，一个是抗原的结合位点，另一个是另一抗原的结合位点)。双抗体的详细描述可以在Holliger P et al.(““Diabodies”:small bivalent and bispecificantibody fragments.”,Proc Natl Acad Sci U S A.1993 Jul 15；90(14):6444-8)中找到。
本文所使用的“单结构域抗体”(也称为“纳米抗体TM”)是由单个单体可变抗体结构域组成的抗体片段。产生单结构域抗体的结构和方法是本领域已知的，例如Holt LJ etal.(”Domain antibodies:proteins for therapy.”Trends Biotechnol.2003 Nov；21(11):484-90)、Saerens D et al.(”Single-domain antibodies as building blocksfor novel therapeutics.”Curr Opin Pharmacol.2008 Oct；8(5):600-8.Epub 2008 Aug22)，和Arbabi Ghahroudi M et al.(“Selection and identification of singledomain antibody fragments from camel heavy-chain antibodies.”FEBS Lett.1997Sep 15；414(3):521-6)。
在本文中与本发明有关的“融合蛋白”不限于特定类型的融合蛋白，只要融合蛋白的各部分通过共价键融合即可。例如，所述融合蛋白的各部分可以通过在一条或多条单多肽链中表达、通过一个或多个二硫键连接、通过化学偶联(优选通过使用点击化学的化学偶联)和/或通过任何其他本领域已知的适合蛋白质连接的共价连接而融合。优选地，所述融合蛋白的各部分通过在一条或多条单多肽链中的表达和/或通过一个或多个二硫键而融合。因此，在本发明的融合蛋白中，抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域，可通过在一条或多条单多肽链中表达、通过一个或多个二硫键连接、通过化学偶联(优选使用点击化学的化学偶联)和/或通过任何其他本领域已知的适合蛋白质连接的共价连接而融合。优选地，所述抗TNFRSF受体抗体或其抗原结合部分与能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域，通过在一条或多条单多肽链中表达和/或通过一个或多个二硫键连接而融合。在本发明的所有实施方案中，其中，能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域包括：
-抗体的抗原结合部分，所述抗原结合部分能与细胞表面的所述结构和/或胞外基质的所述结构结合，其中能与细胞表面和/或胞外基质的所述结构结合的所述抗原结合部分是与一种或多种FcγR类型结合的能力降低的抗原结合部分，所述抗原结合部分是Fab2片段；或
-能结合细胞表面的所述结构和/或胞外基质的所述结构的全长抗体；
非常优选地，所述抗TNFRSF受体抗体或其抗原结合部分与所述结构域通过化学偶联，优选使用点击化学的化学偶联来融合。
通常，就本发明的所有融合蛋白而言，应当理解的是，所述融合蛋白的各部分(例如抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域)可以通过接头序列融合。在这种情况下，本发明的融合蛋白将包含此类接头序列。合适的接头序列是本领域已知的，并且包括例如肽接头，但不限于此。例如，在本发明所述的通过在一条或多条单多肽链中表达而融合的抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域的融合蛋白中，所述一条或多条单多肽链可包含一个或多个肽接头序列，所述肽接头序列位于所述抗TNFRSF受体抗体或其抗原结合部分与能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域之间。
就本发明而言，术语“能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域”是指所述结构域能与不同于FcγR的所述细胞表面的结构和/或所述胞外基质的结构结合。除非另有说明，否则不排除所述结构域还可能包含能与FcγR结合的其他部分(例如Fc结构域)的可能性。
根据本发明，术语“包含”的每次出现可以可选地被术语“由...组成”代替。
方法与技术
通常，除非本文另有定义，否则本发明中使用的方法(例如，克隆方法或与抗体有关的方法)是根据本领域已知的方法进行的，例如，Sambrook et al.(“MolecularCloning:A Laboratory Manual.”,2nd Ed.,Cold Spring Harbor Laboratory Press,ColdSpring Harbor,New York 1989)、Ausubel et al.(“Current Protocols in MolecularBiology.”Greene Publishing Associates and Wiley Interscience；New York 1992)和Harlow and Lane(“Antibodies:A Laboratory Manual”,Cold Spring HarborLaboratory Press,Cold Spring Harbor,New York 1988)中所述的方法，所有其通过引用并入本文。
融合蛋白及其结构域与它们各自的靶蛋白的结合可以通过本领域已知的方法来评估。所述结合优选通过表面等振离子共振测定来评估。
根据本发明，对序列通过使用BLAST算法进行序列比对(参见Altschul et al.(1990)”Basic local alignment search tool.”Journal of Molecular Biology215.p.403-410.；Altschul et al.:(1997)Gapped BLAST and PSI-BLAST:a newgeneration of protein database search programs.NucleicAcids Res.25:3389-3402，其全部通过引用并入本文)。优选地，使用以下参数：最大靶序列10；和字长3；BLOSUM 62矩阵；空位罚分：存在空位11分，延伸1分；条件成分分数矩阵调整。因此，当与序列结合使用时，诸如“同一性”或“相同性”的术语是指通过使用BLAST算法获得的同一性值。
根据本发明，抗体融合蛋白可以通过本领域已知的任何方法产生，包括但不限于在Siegel DL(“Recombinant monoclonal antibody technology.”TransfusClinBiol.2002Jan；9(1):15-22，其通过引用并入本文)中提及的方法。
TNFRSF受体的刺激可以通过本领域已知的任何方法测定。例如，在TNFRSF受体为TNFR2、CD40、CD95、Fn14的情况下，可以通过使用基于细胞的IL8分泌测定法、并随后通过ELISA检测分泌的IL8。可用于此类基于细胞的IL8分泌测定的细胞的非限制性示例为HT1080 CD40转染子(可用于测定CD40刺激)、Widr细胞(可用于测定Fn14刺激)、H1080-细胞(可用于测定CD95刺激)，以及表达TNFR2的HeLa转染子(可用于测定TNFR2刺激)。此类测定的非限制性实施例可以在实施例1-3和相应的图中找到。对于当刺激时具有细胞毒性作用的TNFRSF受体，例如CD95，优选将细胞可渗透的半胱氨酸蛋白酶抑制剂例如ZVAD添加到测定中，以防止诱导细胞死亡。本发明的融合蛋白(例如TRAAFFIAAS)对各种TNFRSF受体的刺激也可以通过任何能检测经典NFκB途径的刺激(例如IκBα的磷酸化和降解、IKK激活、p65的磷酸化和核易位、检测靶基因)或NFκB旁路途径的刺激(例如p100降解、NIK积累)。TRAAFFIAA对CD95和TRAIL死亡受体(DR4、DR5)的刺激作用也可以通过各种测定细胞死亡诱导方法来评估。TRAAFFIA对CD40的刺激可进一步通过测定树突细胞的成熟度来评估。
本发明组合物的制备
根据用于制备药物组合物的已知标准来制备根据本发明的组合物。
例如，所述组合物以其可被适当地储存和施用的方式制备，例如，通过添加药学上可接受的组分，例如载体、赋形剂或稳定剂。
当向患者施用所述药物组合物时，这样的药学上可接受的组分就使用量而言是无毒的。添加到药物组合物中的药学上可接受的组分可以取决于所述药物组合物的具体预期用途和给药途径。
通常，用于本发明的药学上可接受的组分是根据本领域可获得的知识来使用的，例如来自Remington's Pharmaceutical Sciences，Ed.AR Gennaro,20th edition,2000,Williams&Wilkins,PA,USA。
序列
本申请使用的氨基酸序列以单字母氨基酸编码按照N-末端至C-末端的顺序表示。本申请使用的核苷酸序列以标准核酸代码按照5'至3'的顺序表示。星号表示蛋白质序列的末端。带下划线的核酸序列代表限制性核酸内切酶位点。
以下非限制性示例性序列用于本申请的实施例中：
表1:
SEQ ID NO:1:抗CD40-Flag-VH-heavy-full-scTNF80(mu)-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcgagttcacacgggacaaacctgtggctcatgtggtggccaatcatcaggtggaggaacagctggaatggctgagtcagagagcaaacgccctgctggcaaatgggatggacctcaaagacaatcagctcgtggtgcctgccgatggactgtacctggtgtactctcaggtcctgtttaagggacagggatgccccgattacgtgctgctcacccacactgtgtcacgcttcgccatctcataccaggagaaagtcaatctcctctccgccgtgaaatcaccatgtcctaaggatactcccgagggagccgaactgaaaccttggtacgaacccatctacctgggcggcgtgtttcagctggagaaaggcgatcagctctccgccgaagtgaatctgcccaaatacctcaactttagggaatccggacaggtctactttggcgtgattgccctgggaggcggatctggaggaggctctggcgggggatctgggggcggatccgacaaacctgtggcacacgtcgtggcaaaccatcaggtcgaggaacagctcgagtggctgtcacagagggccaatgccctgctggcaaatggaatggatctgaaggataatcagctcgtcgtgcctgccgacggcctctacctcgtctactctcaggtcctctttaagggacagggctgccccgactacgtcctgctcactcataccgtgagtcgcttcgctatttcataccaggaaaaagtcaacctgctgagtgctgtgaaatctccttgccctaaggatacccctgagggagccgaactcaaaccatggtacgagccaatctacctcggaggagtgtttcagctggaaaaaggggatcagctctccgccgaagtcaacctccccaaatacctcaatttccgggaatccggacaggtgtactttggagtcattgccctgggaggcggctctggcgggggatctggaggaggctccggaggaggcagtgacaaacccgtcgctcacgtggtggcaaatcatcaggtcgaggaacagctggaatggctgtctcagagagcaaacgctctcctcgccaatggaatggatctcaaggacaaccagctcgtcgtccctgccgatggactctacctggtctactctcaggtgctctttaagggacagggatgccccgattacgtcctgctcacacacaccgtgtctcgctttgctatttcataccaggagaaagtcaatctgctgtctgccgtcaaatctccttgtccaaaagacacacccgagggagccgaactcaaaccttggtacgagccaatctacctggggggagtgtttcagctggagaagggggatcagctctccgccgaagtgaatctcccaaaatacctcaattttcgggaatccggacaggtctactttggagtgattgccctgtag
SEQ ID NO:19:SEQ ID NO:1表达的蛋白质构建体：
SEQ ID NO:2:抗CD40-Flag-VH-heavy-full-pCR3(hIgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:20:SEQ ID NO:2表达的蛋白质构建体：
SEQ ID NO:3:抗CD40-Flag-VH-heavy-full-scFv-抗CD20-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaAatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagCcaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggcgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaGgggggaccaagctggaaatcaaacgttaa
SEQ ID NO:21:SEQ ID NO:3表达的蛋白质构建体：
SEQ ID NO:4:抗CD40-Flag-VH-heavy-full-scBaff-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:22:SEQ ID NO:4表达的蛋白质构建体：
SEQ ID NO:5:抗CD40-Flag-VL-light-full-pCR3:
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatcgtgatgactcagaacccactgtctctgcctgtgtctctgggggatgaggctagcatttcttgccgctcatctcagtcactggagaactccaatggcaacaccttcctgaattggtttttccagaaacccggccagtcacctcagctgctcatctaccgagtgagcaatcggtttagcggagtgcccgatcgattctctggctccggatctgggaccgactttaccctgaaaatctcacgagtggaggccgaggatctgggagtgtacttctgtctccaggtcacacatgtgccttacacatttggcggcggaacaactctcgaaatcaaaggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:23:SEQ ID NO:5表达的蛋白质构建体：
SEQ ID NO:6:C4-HC-full-IL2(mu)-pCR3(hIgG1)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattgcccggtaccgcccctaccagcagcagcacctctagctctacagccgaggctcaacaacaacaacaacagcaacagcagcagcagcagcacctggaacagctgctgatggacctgcaggaactgctgagccggatggaaaactaccggaacctgaagctgccccggatgctgaccttcaagttctacctgcccaagcaggccaccgagctgaaggatctgcagtgcctggaagatgagctgggccccctgagacacgtgctggatctgacccagagcaagagctttcagctggaagatgccgagaacttcatcagcaacatcagagtgaccgtcgtgaagctgaagggcagcgacaacaccttcgagtgccagttcgacgacgagagcgctaccgtggtggacttcctgcggagatggatcgccttctgccagagcatcatcagcaccagcccccagtaa
SEQ ID NO:24:SEQ ID NO:6表达的蛋白质构建体：
SEQ ID NO:7:C4-HC-heavy-konst-full-scFv-抗CD19-pCR3(N297A)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcgacattcagatgacgcagtctccatcctccatgtctgtatctctgggagacacagtcagcatcacttgccatgcaagtcagggcattagcagtaatatagggtggttgcagcagaaaccagggaaatcatttaagggcctgatctatcatggaaccaacttggaagatggagttccatcaaggttcagtggcagtggatctggagcagattattctctcaccatcagcagcctggaatctgaagattttgcagactattactgtgtacagtatgctcagtttccgtacacgttcggaggggggaccaagctggagctgaaacgtggtggtggtggttctggtggtggtggttctggcggcggcggctccagtggtggtggatcccaggttcagctgcagcaatctggacctgagctggtgaagcctggggcctcagtgaagatttcctgcaaagcttctggctacgcattcagtagctcttggatggactgggtgaagcagaggcctggacagggtcttgagtggattggacggatttatcctggagatggagatactaactacaatgggaagttcaagggcaaggccacactgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacctctgtggactctgcggtctatttctgtgcaaggtccattactacggtagtagggtggtacttcgatgtctggggcgcagggaccacggtcaccgtttcctcctaa
SEQ ID NO:25:SEQ ID NO:7表达的蛋白质构建体：
SEQ ID NO:8:C4-HC-heavy-konst-full-scFv-抗CD20-pCR3(N297A)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggaccctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:26:SEQ ID NO:8表达的蛋白质构建体：
SEQ ID NO:9:C4-HC-heavy-full-pCR3(IgG1)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:27:SEQ ID NO:9表达的蛋白质构建体：
SEQ ID NO:10:C4-HC-heavy-full-scGITRL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcggcggagggagcggacagctggaaactgccaaagaaccctgtatggccaaattcggaccactgcctagcaaatggcagatggctagtagcgaacctccatgtgtgaacaaagtgagcgattggaaactcgagatcctccagaatggactgtacctcatctacggacaggtcgccccaaatgccaattacaatgatgtggccccctttgaagtccggctctacaaaaacaaggatatgatccagaccctcaccaacaaatccaaaatccagaatgtgggcggcacatacgaactccatgtcggcgataccatcgatctcattttcaactctgaacaccaggtgctcaaaaacaacacctactggggaatcatcctgctggcaaaccctcagttcatctccggcggcggctctggcggcggatctggcggagggagtggcggaggctcacagctggagactgctaaagaaccctgtatggcaaaattcgggcccctgccctcaaaatggcagatggcctcctctgaaccaccctgtgtgaacaaagtgagtgattggaaactggaaatcctccagaacggcctctacctcatctacggacaggtggcacccaatgccaattacaacgacgtggcacccttcgaagtgagactgtacaaaaacaaggatatgatccagaccctcaccaacaaatcaaaaatccagaatgtcggagggacatacgaactccatgtgggagacactatcgacctcattttcaattccgaacatcaggtcctgaaaaacaacacttactggggcattattctgctcgccaatccacagtttattagtggaggcgggggatctgggggaggctccggcggagggagtggaggcggatctcaattccaattagagactgctaaggagccctgtatggctaagtttggaccattaccctcaaaatggcaaatggcatcttctgaacctccttgcgtgaataaggtgtctgactggaagctggagatacttcagaatggcttatatttaatttatggccaagtggctcccaatgcaaactacaatgatgtagctccttttgaggtgcggctgtataaaaacaaagacatgatacaaactctaacaaacaaatctaaaatccaaaatgtaggagggacttatgaattgcatgttggggacaccatagacttgatattcaactctgagcatcaggttctaaaaaataatacatactggggtatcattttactagcaaatccccaattcatctcctag
SEQ ID NO:28:SEQ ID NO:10表达的蛋白质构建体：
SEQ ID NO:11:C4-HC-heavy-full-sc(mu)41BBL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcggcggtggaagtggtactgaaccgcgacccgctctcactataaccaccagccctaacttgggaacgcgggagaataacgcggatcaagtaaccccagtttcccatatcgggtgtccaaataccactcaacaaggatcaccggtctttgctaagctcctggcgaaaaaccaagcgtccctctgtaatacaacccttaattggcactcccaagatggggcgggttcttcatacctctcacaggggctccgatacgaagaagacaaaaaggagttggtggtcgactcaccaggactttactatgtattcctggagttgaagctctctcctacattcaccaataccggtcacaaagtacagggttgggtgagccttgtgctccaggcgaagccgcaggtggacgacttcgacaaccttgcactcactgtagagctgttcccttgttccatggagaataaattggtggaccgctcttggagtcagctcctcctcctcaaggcgggtcatcgattgagcgttggccttcgggcatatcttcatggggcgcaagatgcataccgcgattgggaattgtcataccccaacacgacctcttttggcctgtttctggtcaaacccgacaatccgtgggaaggtgggggaagcggagggggttcaggaggaggatctgggggaggttcaactgaaccgaggcccgcgcttactatcacgacttccccaaatctggggactagggaaaacaatgccgatcaggtcactcctgttagtcacattggttgtcctaatacgacccagcaaggctctccggtgtttgccaaactgttggccaaaaatcaggcgtcactttgtaatacaacgctcaactggcatagtcaggatggggccggctcctcatacttgtctcaaggtcttaggtacgaagaagataagaaggagctggtggtagacagccccgggctctactatgtgttcctggagctcaaactgtcaccgacgttcactaacaccggtcataaggtacagggttgggtatccttggtgttgcaagcaaaaccccaggtggacgatttcgataatcttgcgcttactgtagagctctttccatgttcaatggaaaataaactggtcgataggagctggtcccaacttctccttcttaaagctggccatcgcctgagtgttggcctgagagcgtatcttcatggggcgcaggacgcttaccgggattgggaactgtcatacccaaacaccaccagctttgggctcttccttgtaaagccagacaatccgtgggaggggggaggcgggagtgggggcgggtctggagggggcagtggggggggtagtacggagccgcgccccgccctgaccatcacaacgtcacccaatcttgggactcgggagaataacgccgaccaggttacccctgtatcccatatcggttgtcctaatacgacacaacaaggcagtcctgtattcgctaaactcttggcaaaaaaccaggccagtctttgtaatacgacgcttaattggcatagccaggacggtgcgggcagctcctacctttcccaggggctcaggtatgaagaagataagaaagaactcgttgtagacagtcccggattgtattacgtttttttggaactcaagctctctccaaccttcaccaatacgggacataaggtccagggctgggtgagcctcgtactccaggctaagccgcaagttgacgatttcgataatctcgctcttacagtggagttgtttccctgtagtatggagaataagctcgtcgaccggtcttggagccaacttctgctgcttaaggctggtcaccggctcagtgtaggcctccgagcgtatttgcatggggcgcaggacgcctatcgagactgggagctttcctaccctaacacgaccagctttggactcttcttggtgaaacctgacaatccgtgggaataa
SEQ ID NO:29:SEQ ID NO:11表达的蛋白质构建体：
SEQ ID NO:12:C4-HC-heavy-full-(mu)GITRL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcccaactgccatcgagtcctgcatggttaagtttgaactatcatcctcaaaatggcacatgacatctcccaaacctcactgtgtgaatacgacatctgatgggaagctgaagatactgcagagtggcacatatttaatctacggccaagtgattcctgtggataagaaatacataaaagacaatgcccccttcgtagtacagatatataaaaagaatgatgtcctacaaactctaatgaatgattttcaaatcttgcctataggaggggtttatgaactgcatgctggagataacatatatctgaagttcaactctaaagaccatattcagaaaactaacacatactgggggatcatcttaatgcctgatctaccattcatctcttag
SEQ ID NO:30:SEQ ID NO:12表达的蛋白质构建体：
SEQ ID NO:13:C4-LC-light-full-pCR3(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtggatactgctgtagcctggtatcaacaaaaaccagggcaatctcctaaactactgatttactgggcatccacccggcacactggagtccctgatcgcttcacaggcagtggatctgggacagattatactctcaccatcagcagtgtgcaggctgaagacctggcgcgttattactgtcagcaatattatagtgttcctccgacgttcggtggaggcaccaagctgggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:31:SEQ ID NO:13表达的蛋白质构建体：
SEQ ID NO:14:抗CD95(E09)-Flag-VH-full-scFv-抗CD19-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggacattcagatgacgcagtctccatcctccatgtctgtatctctgggagacacagtcagcatcacttgccatgcaagtcagggcattagcagtaatatagggtggttgcagcagaaaccagggaaatcatttaagggcctgatctatcatggaaccaacttggaagatggagttccatcaaggttcagtggcagtggatctggagcagattattctctcaccatcagcagcctggaatctgaagattttgcagactattactgtgtacagtatgctcagtttccgtacacgttcggaggggggaccaagctggagctgaaacgtggtggtggtggttctggtggtggtggttctggcggcggcggctccagtggtggtggatcccaggttcagctgcagcaatctggacctgagctggtgaagcctggggcctcagtgaagatttcctgcaaagcttctggctacgcattcagtagctcttggatggactgggtgaagcagaggcctggacagggtcttgagtggattggacggatttatcctggagatggagatactaactacaatgggaagttcaagggcaaggccacactgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacctctgtggactctgcggtctatttctgtgcaaggtccattactacggtagtagggtggtacttcgatgtctggggcgcagggaccacggtcaccgtttcctcctaa
SEQ ID NO:32:SEQ ID NO:14表达的蛋白质构建体：
SEQ ID NO:15:抗CD95-E09-Flag-VH-heavy-full-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:33:SEQ ID NO:15表达的蛋白质构建体：
SEQ ID NO:16:抗CD95(E09)-Flag-VH-FAB2(1-114)-scFv-抗CD20-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagcaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:34:SEQ ID NO:16表达的蛋白质构建体：
SEQ ID NO:17:抗CD95-E09-Flag-VH-heavy-full-scBaff-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:35:SEQ ID NO:17表达的蛋白质构建体：
SEQ ID NO:18:抗CD95-E09-Flag-VL-light-full-pCR3:
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagagcgtgctgacccagcctcctagcgtgtccgaagcccctagacagaccgtgaccatcagctgctccggcaacagcttcaacatcggcagataccccgtgaactggtatcagcagctgccaggcaaggcccctaaactgctgatctactataacaacctgcggttcagcggagtgtccgaccggttctctggcagcaagtctggcacatctgccagcctggccatccgggatctgctgtctgaggacgaggccgactactactgcagcacctgggacgacaccctgaagggctgggtgttcggcggaggcaccaaagtgacagtgctgggcg gatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:36:SEQ ID NO:18表达的蛋白质构建体：
表2：CDR序列(根据Kabat)。注意，所述重链和轻链CDR序列分别由“-H”和“-L”指示。
SEQ ID NO:96:SEQ ID NO:21所示蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:97:SEQ ID NO:24所示的蛋白质构建体的鼠源IL-2锚定结构域：
SEQ ID NO:98:SEQ ID NO:25所示的蛋白质构建体的scFv-抗CD19锚定结构域：
SEQ ID NO:99:SEQ ID NO:26所示的蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:100:SEQ ID NO:32所示的蛋白质构建体的scFv-抗CD19锚定结构域：
SEQ ID NO:101:SEQ ID NO:34所示的蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:102:SEQ ID NO:19所示的蛋白质构建体的scTNF80(鼠)锚定结构域：
SEQ ID NO:103:SEQ ID NO:22所示的蛋白质构建体的scBaff锚定结构域：
SEQ ID NO:104:SEQ ID NO:28所示的蛋白质构建体的scGITRL锚定结构域：
SEQ ID NO:105:SEQ ID NO:29所示的蛋白质构建体的鼠sc41BBL锚定结构域：
SEQ ID NO:106:SEQ ID NO:30所示的蛋白质构建体的鼠GITRL锚定结构域：
SEQ ID NO:107:人IL-2锚定结构域：
SEQ ID NO:108:抗CD40(G28.5)-VH(1-114)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcctaa
SEQ ID NO:117:SEQ ID NO:108所示的蛋白质构建体：
SEQ ID NO:109:抗CD40(G28.5)-VH(1-114)-scBaff
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:118:SEQ ID NO:109所示的蛋白质构建体
SEQ ID NO:110:抗CD95(E09)-VH(1-114)-scBaff
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:119:SEQ ID NO:110所示的蛋白质构建体
SEQ ID NO:111:抗TNFR2(C4)-IgG2-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctcgagtgctagcaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcggcgtgcacaccttcccagctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaagaatt ccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:120:SEQ ID NO:111所示的蛋白质构建体
SEQ ID NO:112:抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(1F6)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccagattcagctcgtccagtccggacctgaagtgaaaaaacctggcgaaaccgtgaaaatttcctgtaaggcctctggctacacctttaccaactacggcatgaactgggtcaaacaggctcctgggaagggcctgaaatggatgggatggatcaacacctacaccggcgaaccaacatacgccgatgcctttaagggacgctttgccttctctctggaaacttccgcctctactgcttacctccagatcaataacctcaaaaacgaggacaccgccacttacttttgtgctcgggattacggggactacgggatggattactggggacagggaacatctgtgaccgtgtctagcgcttctacaaaggggcctaaactggaggagggcgagtttagcgaggctagagtggatatcgtgctcacacagtctcccgcttctctggctgtctcactgggccagcgagcaacaatctcttgtcgggcttccaaatccgtgtctactagcggctactcttttatgcactggtaccagcagaaacctgggcagcctccaaaactgctcatctacctggcttcaaacctcgaatccggagtgcctgctcgattttctggctctggctccgggaccgactttacactgaacattcatcctgtcgaggaggaggacgctgccacatactactgtcagcattctagggaggtgccatggacatttggcgggggaacaaaactggaaatcaaacggtaa
SEQ ID NO:121:SEQ ID NO:112所示的蛋白质构建体：
SEQ ID NO:113:抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(2H5)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtgcagctggtggaatctggcggcggagtggtgcagcctggcagaagcctgagactgagctgtgccgccagcggcttcaccttcagcagctacatcatgcactgggtgcgccaggcccctggcaagggactggaatgggtggccgtgatcagctacgacggccggaacaagtactacgccgacagcgtgaagggccggttcaccatctcccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgcgggccgaggacaccgccgtgtactactgtgccagagacaccgacggctacgacttcgactattggggccagggcaccctcgtgaccgtgtctagcggaggcggaggatctggcggagggggatcaggcgggggaggctctgaaatcgtgctgacacagagccccgccaccctgtcactgtctccaggcgaaagagccaccctgagctgcagagccagccagagcgtgtccagctacctggcctggtatcagcagaagcccggacaggcccccagactgctgatctacgacgccagcaatcgggccacaggcatccctgccagattttccggctctggcagcggcaccgacttcaccctgacaatcagcagcctggaacccgaggactttgccgtgtattattgccagcagcggaccaactggcccctgacctttggcggaggcaccaaggtggaaatcaaggccagcaccaagggctaa
SEQ ID NO:122:SEQ ID NO:113所示的蛋白质构建体：
SEQ ID NO:114:抗41BB(HBBK4)-IgG1(N297A)-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagcaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:123:SEQ ID NO:114所示的蛋白质构建体：
SEQ ID NO:115:抗41BB(HBBK4)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatcgtgatgactcagagccccgccttcctgtctgtgacccctggcgagaaagtgaccatcacctgtagagccagccagaccatcagcgactacctgcactggtatcagcagaagcccgatcaggcccctaagctgctgattaagtacgcctctcagagcatcagcggcatccccagcagattttctggcagcggctctggcaccgacttcacctttaccatcagctccctggaagccgaggatgccgccacctactattgtcaggacggccacagcttccctccaacctttggacagggcacaaagctggaaatcaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:124:SEQ ID NO:115所示的蛋白质构建体：
SEQ ID NO:116:抗CD95(E09)-IgG1(N297A)-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcntgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcgg atccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:125:SEQ ID NO:116所示的蛋白质构建体
以下如SEQ ID NO 126至143的氨基酸序列是抗4-1BB单克隆抗体HBBK4(其在本申请的非限制性实验中使用)、URELUMAB和UTOMILUMB的CDR序列。注意，术语“CDR1-H”、“CDR2-H”和“CDR3-H”分别指重链CDR1、CDR2和CDR3序列，术语“CDR1-L”、“CDR2-L”和“CDR3-L”分别指轻链CDR1、CDR2和CDR3序列：
SEQ ID NO:126:HBBK4 CDR1-H:
YTFSSYWMH
SEQ ID NO:127:HBBK4 CDR2-H:
EINPGNGHTNYNEKFKS
SEQ ID NO:128:HBBK4 CDR3-H:
SFTTARAFAY
SEQ ID NO:129:HBBK4 CDR1-L:
RASQTISDYLH
SEQ ID NO:130:HBBK4 CDR2-L:
LASQSIS
SEQ ID NO:131:HBBK4 CDR3-L:
QDGHSFPPT
SEQ ID NO:132:URELUMAB CDR1-H:
GSFSGYYWS
SEQ ID NO:133:URELUMAB CDR2-H:
EINHGGYVTYNPSLES
SEQ ID NO:134:URELUMAB CDR3-H:
DYGPGNYDWYFDL
SEQ ID NO:135:URELUMAB CDR1-L:
RASQSVSSYLA
SEQ ID NO:136:URELUMAB CDR2-L:
DASNRAT
SEQ ID NO:137:URELUMAB CDR3-L:
QQRSNWPPALT
SEQ ID NO:138:UTOMILUMAB CDR1-H:
YSFSTYWIS
SEQ ID NO:139:UTOMILUMAB CDR2-H:
KIYPGDSYTNYSPSFQG
SEQ ID NO:140:UTOMILUMAB CDR3-H:
GYGIFDY
SEQ ID NO:141:UTOMILUMAB CDR1-L:
SGDNIGDQYAH
SEQ ID NO:142:UTOMILUMAB CDR2-L:
QDKNRPS
SEQ ID NO:143:UTOMILUMAB CDR3-L:
ATYTGFGSLAV
SEQ ID NO:144:SEQ ID NO:121所示的蛋白质构建体的scFv:CD70(1F6)锚定结构域：
SEQ ID NO:145:SEQ ID NO:122所示的蛋白质构建体的scFv:CD70(2H5)锚定结构域：
SEQ ID NO:146:抗TRAILR2(Cona)-IgG1(N297A)-VH-scFv:CD70(9G2)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctgcaggaatctggccctggcctcgtgaagcctagccagaccctgagcctgacctgtaccgtgtctggcggcagcatcagcagcggcgactacttctggtcctggatcagacagctgcccggcaagggcctggaatggatcggccacatccacaacagcggcaccacctactacaaccccagcctgaagtccagagtgaccatcagcgtggacaccagcaagaagcagttcagcctgcggctgagcagcgtgacagccgccgatacagccgtgtactactgcgccagagacagaggcggcgattactactacggcatggacgtgtggggccagggcaccaccgtgaccgtgtctagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagccccggcaagctcgagcaggtgcagctggtggaatctggcggcggactgatgcagcctggcggctctctgagactgagctgtgccgccagcggcttcacctttagcagcagcgccatgagctgggtgcgccaggctcctggaaagggcctggaatgggtgtccagcatctacagcgacagcagctacacctactacgccgacagcgtgaagtcccggttcaccatcagcaccgacaacgccaagaacaccctgtacctgcagatgaacagcctgaagcccgacgacaccgccgtgtactactgtgccggcagcagcgattacgagggcagctttgcctcttggggccagggcacacaagtgaccgtgtcctccagatctagcaccaagggccccaagctggaagagggcgagttcagcgaggcccaattgcagagcgtcgtgacccagcctcctagcctgtctgcctctcctggaagcagcgtgcggctgacctgtacactgagcagcggcaacagcgtgggcaactacgacatcagctggtatcagcagaaggccggcagcccccccagatacctgctgtactactacagcgattccgtgaagcaccagggcagcggcgtgcccagcagattttccggaagctctgacgccagcgccaacgccggactgctgctgatttctggcctgcagcctgaggacgaggccgactactactgcagcgcctacaagagcggcagccacgtgttcggcggaggcaccaaactgacagtgctgggctaa
SEQ ID NO:157:SEQ ID NO:146所示的蛋白质构建体
SEQ ID NO:147:抗TRAILR2(Cona)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggagatcgtgctgacccagagccctggcaccctgtcactgtctccaggcgagagagccaccctgagctgtagagccagccagggcatcagccggtcttacctggcctggtatcagcagaagcccggccaggctcctagcctgctgatctacggcgccagcagcagagccaccggcatccccgatagattttccggcagcggctccggcaccgacttcaccctgacaatcagcagactggaacccgaggacttcgccgtgtattattgccagcagttcggcagcagcccctggacctttggccagggaacaaaagtgggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:158:SEQ ID NO:147所示的蛋白质构建体
SEQ ID NO:148:抗CD40(C)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctggttcagtctggcgccgaagtgaaaaagcctggcgcctctgtgaaggtgtcctgtacagccagcggcttcaacatcaaggactactacgtgcactgggtcaagcaggcccctggacaaggactggaatggatgggcagaatcgaccccgaggacggcgactctaagtacgcccctaagttccagggcaaagccaccatgaccgccgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcaccaccagctactatgtgggcacctacggctattggggccagggcacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:159:SEQ ID NO:148所示的蛋白质构建体
SEQ ID NO:149:抗CD40(C)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatccagatgacacagagccccagcagcctgtctgccagcgtgggagatagagtgaccatcacctgtagcgccagcagcagcgtgtcctacatgctgtggttccagcagaagcctggcaaggcccctaagctgctgatctacagcacctccaatctggccagcggcgtgccaagcagattttctggctctggcagcggcaccgacttcaccctgaccatatctagcctgcagccagaggacttcgccacctactactgccagcagcggacattctacccctacacctttggcggaggcaccaaggtggaaatcaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:160:SEQ ID NO:149所示的蛋白质构建体
SEQ ID NO:150:抗CD40(C)-Fab2-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctggttcagtctggcgccgaagtgaaaaagcctggcgcctctgtgaaggtgtcctgtacagccagcggcttcaacatcaaggactactacgtgcactgggtcaagcaggcccctggacaaggactggaatggatgggcagaatcgaccccgaggacggcgactctaagtacgcccctaagttccagggcaaagccaccatgaccgccgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcaccaccagctactatgtgggcacctacggctattggggccagggcacactggtcaccgtgtccagcagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:161:SEQ ID NO:150所示的蛋白质构建体
SEQ ID NO:151:抗41BB(HBBK)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:162:SEQ ID NO:151所示的蛋白质构建体
SEQ ID NO:152:抗41BB(HBBK)-Fab2-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:163:SEQ ID NO:152所示的蛋白质构建体
SEQ ID NO:153:抗PD-L1(Ave)-IgG2-VH-scFv:41BB(HBBK)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattcgaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcctcgagtgctagcaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcggcgtgcacaccttcccagctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatctagcacaaagggccccaagctggaagagggcgagtttagcgaggcccaattggacatcgtgatgactcagagccccgccttcctgtctgtgacccctggcgagaaagtgaccatcacctgtagagccagccagaccatcagcgactacctgcactggtatcagcagaagcccgatcaggcccctaagctgctgattaagtacgcctctcagagcatcagcggcatccccagcagattttctggcagcggctctggcaccgacttcacctttaccatcagctccctggaagccgaggatgccgccacctactattgtcaggacggccacagcttccctccaacctttggacagggcacaaagctggaaatcaagtaa
SEQ ID NO:164:SEQ ID NO:153所示的蛋白质构建体
SEQ ID NO:154:抗PD-L1(Ave)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgcttggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:165:SEQ ID NO:154所示的蛋白质构建体
SEQ ID NO:155:抗muCD27-IgG1(N297A)-VH-scFv:Fn14(18D1)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaacttgtgaagcctggcagcagcgtgaagatcagctgtaaagccagcggctacaccttcaccagctacgacatgcactggatcaagcagcagcccggcaaaggcctggaatggatcggctggatctaccccggcaacggcaacaccaagtacaaccagaagttcaacggcaaggccacactgaccgccgacatctctagcagcacagcctacatgcagctgagcagcctgaccagcgaagatagcgccgtgtacttctgcgccaaatggggctacaacaacttcgactactggggccagggcgtgatggtcaccgtgtctagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctggtggaatctggcggcggactggtgcagcctggcggatctctgagactgagctgtgccgccagcggcttcaccttcagcaactactggatgagctgggtgcgccaggcccctggcaaaggactggaatgggtgtccggcatcaacccaggcggcacctctacctactacgccgacagcgtgaagggccggttcaccatcagccgggacaacgccaagaacaccctgtacctgcagatgaacagcctgaagtccgaggacaccgccgtgtactactgcgccaagcacctgggcaactggggcgagtacaattactggggccagggcacacaagtgaccgtgtccagtagatctagcaccaagggccccaagctggaagagggcgagttcagcgaggcccaattgcagagcgccctgacccagcctccaagcgtgtcaggctctcctggcaagaccgtgaccatcagctgtgctggcaccggcggagatgtgggctacagaaacagcgtgtcctggtatcagcagctgcccggcatggcccccaaactgctgatctacgacgtggacaagcgggcctctggcatcaccgacagattcagcggcagcaagagcggcgataccgccagcctgacaatcagcggagtgcagagcgaggacgaggccgactactactgtgccagccagagaagcggaatcgccgccgtgtttggcggaggcacacacctgacagtgctgggctaa
SEQ ID NO:166:SEQ ID NO:155所示的蛋白质构建体
SEQ ID NO:156:抗muCD27-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatccagatgacacagagccctgccagcctgtctgcctctctgggagagacagtgtccatcgattgtctggccagcgagggcatcagcaacgacctggcttggtatcagcagaagtccggcaagagccctcagctgctgatcaacagcgccagcagactggaagatggcgtgcccagcagattttctggctctggcagcggcacccggtacagcctgaagatttctggcatgcagcccgaggacgaggccgaatacttctgcctgcaaagctacagaagcccctggacctttggcggaggcacaaagctggaactgaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:167:SEQ ID NO:156所示的蛋白质构建体
通过以下非限制性实施例进一步说明本发明。
实施例
实施例1：结合TNFR2的抗CD40 IgG1 G28.5的融合蛋白在结合TNFR2后表现出增强的CD40刺激活性。
分析IgG1抗体G28.5的融合蛋白(抗CD40(G28.5)-HC:scTNF80)，所述融合蛋白识别TNFRSF受体CD40，其单链结构域包含过短肽接头(scTNF80(mu)结构域)连接的三个拷贝的鼠源TNF的TNFR2特异性突变体，其中所述拷贝在分子内形成三聚体配体结构域，发明人意外观察到以下情况：
如果用抗CD40(G28.5)-HC:scTNF80刺激表达CD40的细胞或细胞共培养物，所述细胞或细胞共培养物未有FcγR或TNFR2表达，则在两种情况下都会导致没有或极小的CD40活化，例如反映在IL8的产生。相反，在表达FcγR的细胞的存在下，所述分子如预期的那样很好地活化了CD40(参见上述说明)。现在证明，与亲本G28.5抗体不同，抗CD40(G28.5)-HC:scTNF80在缺少FcγR的情况下，如果表达TNFR2，也可以以强烈的CD40活化方式起作用(图6)。显然，所述scTNF80结构域介导的抗CD40(G28.5)-HC:scTNF80与细胞结合的TNFR2的结合具有与Fc结构域介导的G28.5与表达FcγR的细胞的结合一样能产生激动性抗体的效应(图2)。这表明仅仅抗TNFRSF受体抗体或抗体融合蛋白的不依赖抗原结合结构域与细胞的结合足以分别赋予抗体和抗体融合蛋白高激动活性。
实施例2：抗TNFRSF受体CD40、CD95和TNFR2抗体与介导与细胞结合分子结合的蛋白结构域融合的融合蛋白，以不依赖FcγR的方式作为强激动剂。
根据仅仅抗TNFRSF受体抗体或抗体融合蛋白的不依赖抗原结合结构域的与细胞的结合足以分别赋予抗体和抗体融合蛋白高激动活性的想法，发明人还能证实识别TNFRSF受体CD40、CD95和TNFR2的抗体结构域与介导与细胞结合分子结合的蛋白结构域融合的其他抗体融合蛋白，能以不依赖FcγR的方式作为强激动剂。有关这些示例的详细信息，请参见图4、5、8和9，包括图例。
实施例3：抗TNFRSF受体CD40、CD95和TNFR2的抗体与靶向细胞表面抗原的scFv结构域的融合蛋白以不依赖FcγR的方式作为强激动剂。
另外，发明人研究了抗TNFRSF受体CD40、CD95和TNFR2的抗体与靶向细胞表面抗原的scFv结构域的融合蛋白，其分别靶向CD20(图10、11和12)和CD19(图7和12)。已发现这些融合蛋白以不依赖FcγR的方式起强激动剂的作用。有关这些示例的详细信息，请参见引用的图及其图例。
实施例4：抗TNFRSF受体CD40和CD95与scBaff的抗体融合蛋白以不依赖FcγR的方式作为强激动剂。
另外，发明人用scBaff研究了抗TNFRSF受体CD40(图13、14和15)和CD95(图16、17和18)的抗体或抗体片段的融合蛋白。已经发现，这些融合蛋白以不依赖FcγR的方式作为CD40和CD95的强激动剂，只要所述融合蛋白具有结合可被融合蛋白的scBaff结构域识别的表达细胞表面受体的可能性。这些示例的详细信息在上述图及其图例中给出。
实施例5：抗TNFRSF受体CD40、CD95、TNFR2和4-1BB的抗体的scFv融合蛋白以不依赖FcγR、scFv依赖的方式作为强激动剂。
此外，发明人研究了抗TNFRSF受体TNFR2(图19和20)、CD40(图21和22)、CD95(图21和22)和4-1BB(图21和22)的抗体的scFv融合蛋白。使用的scFv片段是scFv:CD20(图19、21和22)、scFv:CD70(2H5)(图20)和scFv:CD70(1F6)(图20)。发现这些融合蛋白作为被抗体部分(TNFR2、CD40、CD95、4-1BB)以不依赖FcγR的方式识别的TNFRSF受体的强激动剂，只要所述融合蛋白可以通过其scFv结构域结合被scFv结构域识别的细胞表面抗原(CD20、CD70)。这些示例的详细信息在上述图及其图例中给出。
实施例6：TNFRSF受体TRAILR2(DR5)的scFv融合蛋白以不依赖FcγR、scFv依赖的方式作为强激动剂。
此外，发明人研究了抗TNFRSF受体TRAILR2的抗体(Conatumumab)的scFv融合蛋白，也称为DR5(图23)。所使用的scFv片段是scFv:CD70。发现所述融合蛋白被抗体部分(TRAILR2)以不依赖FcγR的方式识别的TNFRSF受体的强激动剂，只要所述融合蛋白具有通过其scFv结构域与可被scFv结构域识别的细胞表面抗原CD70结合的可能性。
实施例7：抗抗原呈递细胞刺激的TNFRSF受体CD40或T细胞刺激的TNFRSF受体4-1BB的抗体的融合蛋白，其具有来自阻断PD-1L和PD-1相互作用的检查点抑制剂Avelumab的scFv作为锚定结构域，以不依赖FcγR、scFv:PD-1L依赖的方式起强激动剂作用。发明人还研究了抗TNFRSF受体CD40(图24)和4-1BB(图25)的抗体或抗体片段与scFv:PD-1L的融合蛋白，所述scFv:PD-1L是检查点抑制剂Avelumab的scFv，其中Avelumab为一种阻断PD-1L与PD-1结合的抗体。发现这些融合蛋白可以以不依赖FcγR的方式作为CD40和4-1BB的强激动剂，只要所述融合蛋白可能与被融合蛋白的scFv:PD-1L结构域识别的细胞表面表达的PD-1L结合。因此，这些融合蛋白是结合了免疫检查点阻断与免疫检查点阻断依赖性限制性激活免疫刺激因子TNFRSF受体的双功能抗体示例。这些示例的详细信息在上述图及其图例中给出。
实施例8：当与锚定于细胞表面抗原的完整抗体融合时，对TNFRSF受体Fn14和4-1BB特异的scFv以不依赖FcγR的方式作为强激动剂。另外，发明人用识别PD-1L或鼠CD27的抗体研究了抗TNFRSF受体4-1BB(图26)和Fn14(图27)的scFv融合蛋白。已经发现，只要所述融合蛋白的抗体部分有可能结合其细胞表面表达的抗原PD-1L和鼠CD27时，所述融合蛋白以不依赖FcγR的方式作为4-1BB和Fn14的强激动剂。这些示例展示了完整的抗体可以用作锚定结构域，以使抗TNFRSF受体抗体片段(在此为scFv)能作为具有锚定依赖性活性的激动剂。这些示例的详细信息在上述图及其图例中给出。
实施例结论
对于用于不依赖FcγR和抗原结合域的细胞相互作用，并且与抗体融合的结构域，在下文中也称为固定结构域，没有特别限制。因此，本发明人能生产利用IL2配体或scTNFSF配体进行细胞结合的TRAAFFIAA，但也可以产生出于所述目的使用异源scFv抗体片段的TRAAFFIAA。为了获得激动活性，所述抗原结合结构域以不依赖FcγR和抗原结合域的方式结合细胞似乎也是足够的。例如，已经通过与CD20特异性scFv融合进行基因工程改造的CD95特异性抗体E09的Fab2片段(图11)，显示出依赖CD20的强烈增加的TNFRSF受体刺激活性。
从本发明人的工作中，立即可显而易见的是，使用在TRAAFFIAA中使用的经适当选择的固定化结构域，可以在体内获得限于靶结构的局部激动效应。因此，有可能避免可能限制治疗的全身副作用。因此，TRAAFFIAA不仅可以有利地以不依赖FcγR的方式用抗体刺激TNFRSF受体，而且还开拓了包括临床应用的新的应用领域。
工业适用性
根据本发明，所述抗体融合蛋白及其组合物可以工业化生产并作为商业产品出售，例如作为药物。因此，本发明在工业上是可应用的。
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SEQUENCE LISTING
<110> 尤利乌斯·马克西米利安维尔茨堡大学
<120> 具有非FcγR依赖性激动活性的肿瘤坏死因子（TNF）受体超家族（TNFRSF）
受体-激活抗体融合蛋白（具有非FcγR依赖性激动活性的TNFRSF受体-激活
抗体融合蛋白；TRAAFFIAA）
<130> P20113345WP
<150> EP 17210849.0
<151> 2017-12-28
<150> EP 18189937.8
<151> 2018-08-21
<160> 167
<170> PatentIn version 3.5
<210> 1
<211> 2892
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scTNF80(mu)-pCR3 (N297A)
<400> 1
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca aggaattcga gttcacacgg gacaaacctg tggctcatgt ggtggccaat 1500
catcaggtgg aggaacagct ggaatggctg agtcagagag caaacgccct gctggcaaat 1560
gggatggacc tcaaagacaa tcagctcgtg gtgcctgccg atggactgta cctggtgtac 1620
tctcaggtcc tgtttaaggg acagggatgc cccgattacg tgctgctcac ccacactgtg 1680
tcacgcttcg ccatctcata ccaggagaaa gtcaatctcc tctccgccgt gaaatcacca 1740
tgtcctaagg atactcccga gggagccgaa ctgaaacctt ggtacgaacc catctacctg 1800
ggcggcgtgt ttcagctgga gaaaggcgat cagctctccg ccgaagtgaa tctgcccaaa 1860
tacctcaact ttagggaatc cggacaggtc tactttggcg tgattgccct gggaggcgga 1920
tctggaggag gctctggcgg gggatctggg ggcggatccg acaaacctgt ggcacacgtc 1980
gtggcaaacc atcaggtcga ggaacagctc gagtggctgt cacagagggc caatgccctg 2040
ctggcaaatg gaatggatct gaaggataat cagctcgtcg tgcctgccga cggcctctac 2100
ctcgtctact ctcaggtcct ctttaaggga cagggctgcc ccgactacgt cctgctcact 2160
cataccgtga gtcgcttcgc tatttcatac caggaaaaag tcaacctgct gagtgctgtg 2220
aaatctcctt gccctaagga tacccctgag ggagccgaac tcaaaccatg gtacgagcca 2280
atctacctcg gaggagtgtt tcagctggaa aaaggggatc agctctccgc cgaagtcaac 2340
ctccccaaat acctcaattt ccgggaatcc ggacaggtgt actttggagt cattgccctg 2400
ggaggcggct ctggcggggg atctggagga ggctccggag gaggcagtga caaacccgtc 2460
gctcacgtgg tggcaaatca tcaggtcgag gaacagctgg aatggctgtc tcagagagca 2520
aacgctctcc tcgccaatgg aatggatctc aaggacaacc agctcgtcgt ccctgccgat 2580
ggactctacc tggtctactc tcaggtgctc tttaagggac agggatgccc cgattacgtc 2640
ctgctcacac acaccgtgtc tcgctttgct atttcatacc aggagaaagt caatctgctg 2700
tctgccgtca aatctccttg tccaaaagac acacccgagg gagccgaact caaaccttgg 2760
tacgagccaa tctacctggg gggagtgttt cagctggaga agggggatca gctctccgcc 2820
gaagtgaatc tcccaaaata cctcaatttt cgggaatccg gacaggtcta ctttggagtg 2880
attgccctgt ag 2892
<210> 2
<211> 1455
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-pCR3 (hIgG1)
<400> 2
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca agtaa 1455
<210> 3
<211> 2189
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scFv-抗CD20-pCR3 (N297A)
<400> 3
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca aggaattcca ggtacaactg cagcagcctg gggctgagct ggtgaagcct 1500
ggggcctcag tgaagatgtc ctgcaaggct tctggctaca catttaccag ttacaatatg 1560
cactgggtaa aacagacacc tggtcggggc ctggaatgga ttggagctat ttatcccgga 1620
aatggtgata cttcctacaa tcagaagttc aaaggcaagg ccacattgac tgcagacaaa 1680
tcctccagca cagcctacat gcagctcagc agcctgacat ctgaggactc tgcggtctat 1740
tactgtgcaa gatcgactta ctacggcggt gactggtact tcaatgtctg gggcgcaggg 1800
accacggtca ccgtctcttc aggaggaggc ggatccggcg gaggcggaag cggtggcgga 1860
ggctctcaaa ttgttctctc ccagtctcca gcaatcctgt ctgcatctcc aggggagaag 1920
gtcacaatga cttgcagggc cagctcaagt gtaagttaca tccactggtt ccagcagaag 1980
ccaggatcct cccccaaacc ctggatttat gccacatcca acctggcttc tggagtccct 2040
gttcgcttca gtggcagtgg gtctgggact tcttactctc tcacaatcag cagagtggag 2100
gcgaagatgc tgccacttat tactgccagc agtggactag taacccaccc acgttcggag 2160
gggggaccaa gctggaaatc aaacgttaa 2189
<210> 4
<211> 2874
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scBaff-pCR3 (N297A)
<400> 4
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca agctcgaggg acccgaggaa actgtgactc aggactgtct ccagctcatt 1500
gccgatagtg aaacccctac catccagaaa ggctcttaca ccttcgtgcc atggctgctg 1560
tcattcaaac ggggatctgc tctggaggag aaggaaaaca aaatcctggt caaggaaacc 1620
ggctacttct tcatctacgg ccaggtcctc tacaccgaca aaacatacgc tatggggcat 1680
ctcattcagc ggaaaaaagt ccacgtgttc ggcgacgaac tctctctcgt gacactgttc 1740
cggtgtattc agaacatgcc cgagactctg cccaataata gctgctactc tgctggcatt 1800
gccaaactgg aggagggcga cgaactccag ctggctattc ctagggaaaa tgcccagatt 1860
agcctggacg gggatgtgac attttttggc gccctgaaac tgctgggagg cggagggagt 1920
ggcgggggag gctctggacc tgaggaaact gtgacccagg attgtctcca gctcattgcc 1980
gatagtgaga ctcctaccat tcagaaggga tcttacacct ttgtgccttg gctgctgtct 2040
ttcaaacggg gctctgctct ggaggaaaag gagaacaaaa ttctggtcaa agagactggc 2100
tacttcttca tctacggcca ggtgctgtac accgacaaaa catacgccat gggccatctc 2160
attcagcgga aaaaagtcca cgtgttcggc gacgaactct ctctcgtgac actgttccgg 2220
tgtatccaga acatgcccga gacactgccc aataatagct gctactctgc cggcattgct 2280
aaactggagg agggggacga actccagctg gctattccta gggaaaatgc ccagatttct 2340
ctcgatgggg atgtgacatt cttcggggcc ctcaaactgc tgggaggcgg cggatctggc 2400
ggaggcggga gtcaattcgc agcaggtcca gaagaaacag tcactcaaga ctgcttgcaa 2460
ctgattgcag acagtgaaac accaactata caaaaaggat cttacacatt tgttccatgg 2520
cttctcagct ttaaaagggg aagtgcccta gaagaaaaag agaataaaat attggtcaaa 2580
gaaactggtt acttttttat atatggtcag gttttatata ctgataagac ctacgccatg 2640
ggacatctaa ttcagaggaa gaaggtccat gtctttgggg atgaattgag tctggtgact 2700
ttgtttcgat gtattcaaaa tatgcctgaa acactaccca ataattcctg ctattcagct 2760
ggcattgcaa aactggaaga aggagatgaa ctccaacttg caataccaag agaaaatgca 2820
caaatatcac tggatggaga tgtcacattt tttggtgcat tgaaactgct gtga 2874
<210> 5
<211> 789
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VL-light-full-pCR3
<400> 5
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
gtgatgactc agaacccact gtctctgcct gtgtctctgg gggatgaggc tagcatttct 180
tgccgctcat ctcagtcact ggagaactcc aatggcaaca ccttcctgaa ttggtttttc 240
cagaaacccg gccagtcacc tcagctgctc atctaccgag tgagcaatcg gtttagcgga 300
gtgcccgatc gattctctgg ctccggatct gggaccgact ttaccctgaa aatctcacga 360
gtggaggccg aggatctggg agtgtacttc tgtctccagg tcacacatgt gccttacaca 420
tttggcggcg gaacaactct cgaaatcaaa ggatccgaaa tcaagcgtac ggtggccgct 480
cccagcgtgt tcatcttccc acctagcgac gagcagctga agtccggcac agcctctgtc 540
gtgtgcctgc tgaacaactt ctacccccgc gaggccaagg tgcagtggaa ggtggacaat 600
gccctgcaga gcggcaacag ccaggaaagc gtgaccgagc aggacagcaa ggactccacc 660
tacagcctga gcagcaccct gaccctgagc aaggccgact acgagaagca caaggtgtac 720
gcctgcgaag tgacccacca gggcctgtct agccccgtga ccaagagctt caaccggggc 780
gagtgctaa 789
<210> 6
<211> 1908
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-full-IL2(mu)-pCR3 (hIgG1) (无 Flag)
<400> 6
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctct agcgccagca caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgacagtg tcctggaact ctggcgccct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
actgtgccca gcagcagcct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccctga actgctgggc ggaccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcccagaga ggaacagtac aacagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgcctcc cagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta cccctccgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattgc ccggtaccgc ccctaccagc agcagcacct ctagctctac agccgaggct 1500
caacaacaac aacaacagca acagcagcag cagcagcacc tggaacagct gctgatggac 1560
ctgcaggaac tgctgagccg gatggaaaac taccggaacc tgaagctgcc ccggatgctg 1620
accttcaagt tctacctgcc caagcaggcc accgagctga aggatctgca gtgcctggaa 1680
gatgagctgg gccccctgag acacgtgctg gatctgaccc agagcaagag ctttcagctg 1740
gaagatgccg agaacttcat cagcaacatc agagtgaccg tcgtgaagct gaagggcagc 1800
gacaacacct tcgagtgcca gttcgacgac gagagcgcta ccgtggtgga cttcctgcgg 1860
agatggatcg ccttctgcca gagcatcatc agcaccagcc cccagtaa 1908
<210> 7
<211> 2199
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-konst-full-scFv-抗CD19-pCR3 (N297A) (无Flag)
<400> 7
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg acattcagat gacgcagtct ccatcctcca tgtctgtatc tctgggagac 1500
acagtcagca tcacttgcca tgcaagtcag ggcattagca gtaatatagg gtggttgcag 1560
cagaaaccag ggaaatcatt taagggcctg atctatcatg gaaccaactt ggaagatgga 1620
gttccatcaa ggttcagtgg cagtggatct ggagcagatt attctctcac catcagcagc 1680
ctggaatctg aagattttgc agactattac tgtgtacagt atgctcagtt tccgtacacg 1740
ttcggagggg ggaccaagct ggagctgaaa cgtggtggtg gtggttctgg tggtggtggt 1800
tctggcggcg gcggctccag tggtggtgga tcccaggttc agctgcagca atctggacct 1860
gagctggtga agcctggggc ctcagtgaag atttcctgca aagcttctgg ctacgcattc 1920
agtagctctt ggatggactg ggtgaagcag aggcctggac agggtcttga gtggattgga 1980
cggatttatc ctggagatgg agatactaac tacaatggga agttcaaggg caaggccaca 2040
ctgactgcag acaaatcctc cagcacagcc tacatgcagc tcagcagcct gacctctgtg 2100
gactctgcgg tctatttctg tgcaaggtcc attactacgg tagtagggtg gtacttcgat 2160
gtctggggcg cagggaccac ggtcaccgtt tcctcctaa 2199
<210> 8
<211> 2181
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-konst-full-scFv-抗CD20-pCR3 (N297A) (无Flag)
<400> 8
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc aggtacaact gcagcagcct ggggctgagc tggtgaagcc tggggcctca 1500
gtgaagatgt cctgcaaggc ttctggctac acatttacca gttacaatat gcactgggta 1560
aaacagacac ctggtcgggg cctggaatgg attggagcta tttatcccgg aaatggtgat 1620
acttcctaca atcagaagtt caaaggcaag gccacattga ctgcagacaa atcctccagc 1680
acagcctaca tgcagctcag cagcctgaca tctgaggact ctgcggtcta ttactgtgca 1740
agatcgactt actacggcgg tgactggtac ttcaatgtct ggggcgcagg gaccacggtc 1800
accgtctctt caggaggagg cggatccggc ggaggcggaa gcggtggcgg aggctctcaa 1860
attgttctct cccagtctcc agcaatcctg tctgcatctc caggggagaa ggtcacaatg 1920
acttgcaggg ccagctcaag tgtaagttac atccactggt tccagcagaa gccaggaccc 1980
tcccccaaac cctggattta tgccacatcc aacctggctt ctggagtccc tgttcgcttc 2040
agtggcagtg ggtctgggac ttcttactct ctcacaatca gcagagtgga ggctgaagat 2100
gctgccactt attactgcca gcagtggact agtaacccac ccacgttcgg aggggggacc 2160
aagctggaaa tcaaacgtta a 2181
<210> 9
<211> 1446
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-pCR3 (IgG1) (无Flag)
<400> 9
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctct agcgccagca caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgacagtg tcctggaact ctggcgccct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
actgtgccca gcagcagcct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccctga actgctgggc ggaccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcccagaga ggaacagtac aacagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgcctcc cagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta cccctccgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aagtaa 1446
<210> 10
<211> 2724
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-scGITRL-pCR3 (N297A)
<400> 10
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg gcggagggag cggacagctg gaaactgcca aagaaccctg tatggccaaa 1500
ttcggaccac tgcctagcaa atggcagatg gctagtagcg aacctccatg tgtgaacaaa 1560
gtgagcgatt ggaaactcga gatcctccag aatggactgt acctcatcta cggacaggtc 1620
gccccaaatg ccaattacaa tgatgtggcc ccctttgaag tccggctcta caaaaacaag 1680
gatatgatcc agaccctcac caacaaatcc aaaatccaga atgtgggcgg cacatacgaa 1740
ctccatgtcg gcgataccat cgatctcatt ttcaactctg aacaccaggt gctcaaaaac 1800
aacacctact ggggaatcat cctgctggca aaccctcagt tcatctccgg cggcggctct 1860
ggcggcggat ctggcggagg gagtggcgga ggctcacagc tggagactgc taaagaaccc 1920
tgtatggcaa aattcgggcc cctgccctca aaatggcaga tggcctcctc tgaaccaccc 1980
tgtgtgaaca aagtgagtga ttggaaactg gaaatcctcc agaacggcct ctacctcatc 2040
tacggacagg tggcacccaa tgccaattac aacgacgtgg cacccttcga agtgagactg 2100
tacaaaaaca aggatatgat ccagaccctc accaacaaat caaaaatcca gaatgtcgga 2160
gggacatacg aactccatgt gggagacact atcgacctca ttttcaattc cgaacatcag 2220
gtcctgaaaa acaacactta ctggggcatt attctgctcg ccaatccaca gtttattagt 2280
ggaggcgggg gatctggggg aggctccggc ggagggagtg gaggcggatc tcaattccaa 2340
ttagagactg ctaaggagcc ctgtatggct aagtttggac cattaccctc aaaatggcaa 2400
atggcatctt ctgaacctcc ttgcgtgaat aaggtgtctg actggaagct ggagatactt 2460
cagaatggct tatatttaat ttatggccaa gtggctccca atgcaaacta caatgatgta 2520
gctccttttg aggtgcggct gtataaaaac aaagacatga tacaaactct aacaaacaaa 2580
tctaaaatcc aaaatgtagg agggacttat gaattgcatg ttggggacac catagacttg 2640
atattcaact ctgagcatca ggttctaaaa aataatacat actggggtat cattttacta 2700
gcaaatcccc aattcatctc ctag 2724
<210> 11
<211> 3411
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-sc(mu)41BBL-pCR3 (N297A)
<400> 11
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg gcggtggaag tggtactgaa ccgcgacccg ctctcactat aaccaccagc 1500
cctaacttgg gaacgcggga gaataacgcg gatcaagtaa ccccagtttc ccatatcggg 1560
tgtccaaata ccactcaaca aggatcaccg gtctttgcta agctcctggc gaaaaaccaa 1620
gcgtccctct gtaatacaac ccttaattgg cactcccaag atggggcggg ttcttcatac 1680
ctctcacagg ggctccgata cgaagaagac aaaaaggagt tggtggtcga ctcaccagga 1740
ctttactatg tattcctgga gttgaagctc tctcctacat tcaccaatac cggtcacaaa 1800
gtacagggtt gggtgagcct tgtgctccag gcgaagccgc aggtggacga cttcgacaac 1860
cttgcactca ctgtagagct gttcccttgt tccatggaga ataaattggt ggaccgctct 1920
tggagtcagc tcctcctcct caaggcgggt catcgattga gcgttggcct tcgggcatat 1980
cttcatgggg cgcaagatgc ataccgcgat tgggaattgt cataccccaa cacgacctct 2040
tttggcctgt ttctggtcaa acccgacaat ccgtgggaag gtgggggaag cggagggggt 2100
tcaggaggag gatctggggg aggttcaact gaaccgaggc ccgcgcttac tatcacgact 2160
tccccaaatc tggggactag ggaaaacaat gccgatcagg tcactcctgt tagtcacatt 2220
ggttgtccta atacgaccca gcaaggctct ccggtgtttg ccaaactgtt ggccaaaaat 2280
caggcgtcac tttgtaatac aacgctcaac tggcatagtc aggatggggc cggctcctca 2340
tacttgtctc aaggtcttag gtacgaagaa gataagaagg agctggtggt agacagcccc 2400
gggctctact atgtgttcct ggagctcaaa ctgtcaccga cgttcactaa caccggtcat 2460
aaggtacagg gttgggtatc cttggtgttg caagcaaaac cccaggtgga cgatttcgat 2520
aatcttgcgc ttactgtaga gctctttcca tgttcaatgg aaaataaact ggtcgatagg 2580
agctggtccc aacttctcct tcttaaagct ggccatcgcc tgagtgttgg cctgagagcg 2640
tatcttcatg gggcgcagga cgcttaccgg gattgggaac tgtcataccc aaacaccacc 2700
agctttgggc tcttccttgt aaagccagac aatccgtggg aggggggagg cgggagtggg 2760
ggcgggtctg gagggggcag tggggggggt agtacggagc cgcgccccgc cctgaccatc 2820
acaacgtcac ccaatcttgg gactcgggag aataacgccg accaggttac ccctgtatcc 2880
catatcggtt gtcctaatac gacacaacaa ggcagtcctg tattcgctaa actcttggca 2940
aaaaaccagg ccagtctttg taatacgacg cttaattggc atagccagga cggtgcgggc 3000
agctcctacc tttcccaggg gctcaggtat gaagaagata agaaagaact cgttgtagac 3060
agtcccggat tgtattacgt ttttttggaa ctcaagctct ctccaacctt caccaatacg 3120
ggacataagg tccagggctg ggtgagcctc gtactccagg ctaagccgca agttgacgat 3180
ttcgataatc tcgctcttac agtggagttg tttccctgta gtatggagaa taagctcgtc 3240
gaccggtctt ggagccaact tctgctgctt aaggctggtc accggctcag tgtaggcctc 3300
cgagcgtatt tgcatggggc gcaggacgcc tatcgagact gggagctttc ctaccctaac 3360
acgaccagct ttggactctt cttggtgaaa cctgacaatc cgtgggaata a 3411
<210> 12
<211> 1836
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-(mu)GITRL-pCR3 (N297A)
<400> 12
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc caactgccat cgagtcctgc atggttaagt ttgaactatc atcctcaaaa 1500
tggcacatga catctcccaa acctcactgt gtgaatacga catctgatgg gaagctgaag 1560
atactgcaga gtggcacata tttaatctac ggccaagtga ttcctgtgga taagaaatac 1620
ataaaagaca atgccccctt cgtagtacag atatataaaa agaatgatgt cctacaaact 1680
ctaatgaatg attttcaaat cttgcctata ggaggggttt atgaactgca tgctggagat 1740
aacatatatc tgaagttcaa ctctaaagac catattcaga aaactaacac atactggggg 1800
atcatcttaa tgcctgatct accattcatc tcttag 1836
<210> 13
<211> 735
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-LC-light-full-pCR3 (无Flag)
<400> 13
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggacatt gtgatgaccc agtctcacaa attcatgtcc 120
acatcagtag gagacagggt cagcatcacc tgcaaggcca gtcaggatgt ggatactgct 180
gtagcctggt atcaacaaaa accagggcaa tctcctaaac tactgattta ctgggcatcc 240
acccggcaca ctggagtccc tgatcgcttc acaggcagtg gatctgggac agattatact 300
ctcaccatca gcagtgtgca ggctgaagac ctggcgcgtt attactgtca gcaatattat 360
agtgttcctc cgacgttcgg tggaggcacc aagctgggat ccgaaatcaa gcgtacggtg 420
gccgctccca gcgtgttcat cttcccacct agcgacgagc agctgaagtc cggcacagcc 480
tctgtcgtgt gcctgctgaa caacttctac ccccgcgagg ccaaggtgca gtggaaggtg 540
gacaatgccc tgcagagcgg caacagccag gaaagcgtga ccgagcagga cagcaaggac 600
tccacctaca gcctgagcag caccctgacc ctgagcaagg ccgactacga gaagcacaag 660
gtgtacgcct gcgaagtgac ccaccagggc ctgtctagcc ccgtgaccaa gagcttcaac 720
cggggcgagt gctaa 735
<210> 14
<211> 2268
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-Flag-VH-full-scFv-抗CD19-pCR3 (IgG1)
<400> 14
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc agatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agctcgagga cattcagatg acgcagtctc catcctccat gtctgtatct 1560
ctgggagaca cagtcagcat cacttgccat gcaagtcagg gcattagcag taatataggg 1620
tggttgcagc agaaaccagg gaaatcattt aagggcctga tctatcatgg aaccaacttg 1680
gaagatggag ttccatcaag gttcagtggc agtggatctg gagcagatta ttctctcacc 1740
atcagcagcc tggaatctga agattttgca gactattact gtgtacagta tgctcagttt 1800
ccgtacacgt tcggaggggg gaccaagctg gagctgaaac gtggtggtgg tggttctggt 1860
ggtggtggtt ctggcggcgg cggctccagt ggtggtggat cccaggttca gctgcagcaa 1920
tctggacctg agctggtgaa gcctggggcc tcagtgaaga tttcctgcaa agcttctggc 1980
tacgcattca gtagctcttg gatggactgg gtgaagcaga ggcctggaca gggtcttgag 2040
tggattggac ggatttatcc tggagatgga gatactaact acaatgggaa gttcaagggc 2100
aaggccacac tgactgcaga caaatcctcc agcacagcct acatgcagct cagcagcctg 2160
acctctgtgg actctgcggt ctatttctgt gcaaggtcca ttactacggt agtagggtgg 2220
tacttcgatg tctggggcgc agggaccacg gtcaccgttt cctcctaa 2268
<210> 15
<211> 1515
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VH-heavy-full-pCR3 (IgG1)
<400> 15
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc agatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agtaa 1515
<210> 16
<211> 1602
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-Flag-VH-FAB2(1-114)-scFv-抗CD20-pCR3 (IgG1)
<400> 16
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgccctcgag caggtacaac tgcagcagcc tggggctgag 900
ctggtgaagc ctggggcctc agtgaagatg tcctgcaagg cttctggcta cacatttacc 960
agttacaata tgcactgggt aaaacagaca cctggtcggg gcctggaatg gattggagct 1020
atttatcccg gaaatggtga tacttcctac aatcagaagt tcaaaggcaa ggccacattg 1080
actgcagaca aatcctccag cacagcctac atgcagctca gcagcctgac atctgaggac 1140
tctgcggtct attactgtgc aagatcgact tactacggcg gtgactggta cttcaatgtc 1200
tggggcgcag ggaccacggt caccgtctct tcaggaggag gcggatccgg cggaggcgga 1260
agcggtggcg gaggctctca aattgttctc tcccagtctc cagcaatcct gtctgcatct 1320
ccaggggaga aggtcacaat gacttgcagg gccagctcaa gtgtaagtta catccactgg 1380
ttccagcaga agccaggatc ctcccccaaa ccctggattt atgccacatc caacctggct 1440
tctggagtcc ctgttcgctt cagtggcagt gggtctggga cttcttactc tctcacaatc 1500
agcagagtgg aggctgaaga tgctgccact tattactgcc agcagtggac tagtaaccca 1560
cccacgttcg gaggggggac caagctggaa atcaaacgtt aa 1602
<210> 17
<211> 2934
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VH-heavy-full-scBaff-pCR3 (N297A)
<400> 17
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatccagca gcgcctctac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agctcgaggg acccgaggaa actgtgactc aggactgtct ccagctcatt 1560
gccgatagtg aaacccctac catccagaaa ggctcttaca ccttcgtgcc atggctgctg 1620
tcattcaaac ggggatctgc tctggaggag aaggaaaaca aaatcctggt caaggaaacc 1680
ggctacttct tcatctacgg ccaggtcctc tacaccgaca aaacatacgc tatggggcat 1740
ctcattcagc ggaaaaaagt ccacgtgttc ggcgacgaac tctctctcgt gacactgttc 1800
cggtgtattc agaacatgcc cgagactctg cccaataata gctgctactc tgctggcatt 1860
gccaaactgg aggagggcga cgaactccag ctggctattc ctagggaaaa tgcccagatt 1920
agcctggacg gggatgtgac attttttggc gccctgaaac tgctgggagg cggagggagt 1980
ggcgggggag gctctggacc tgaggaaact gtgacccagg attgtctcca gctcattgcc 2040
gatagtgaga ctcctaccat tcagaaggga tcttacacct ttgtgccttg gctgctgtct 2100
ttcaaacggg gctctgctct ggaggaaaag gagaacaaaa ttctggtcaa agagactggc 2160
tacttcttca tctacggcca ggtgctgtac accgacaaaa catacgccat gggccatctc 2220
attcagcgga aaaaagtcca cgtgttcggc gacgaactct ctctcgtgac actgttccgg 2280
tgtatccaga acatgcccga gacactgccc aataatagct gctactctgc cggcattgct 2340
aaactggagg agggggacga actccagctg gctattccta gggaaaatgc ccagatttct 2400
ctcgatgggg atgtgacatt cttcggggcc ctcaaactgc tgggaggcgg cggatctggc 2460
ggaggcggga gtcaattcgc agcaggtcca gaagaaacag tcactcaaga ctgcttgcaa 2520
ctgattgcag acagtgaaac accaactata caaaaaggat cttacacatt tgttccatgg 2580
cttctcagct ttaaaagggg aagtgcccta gaagaaaaag agaataaaat attggtcaaa 2640
gaaactggtt acttttttat atatggtcag gttttatata ctgataagac ctacgccatg 2700
ggacatctaa ttcagaggaa gaaggtccat gtctttgggg atgaattgag tctggtgact 2760
ttgtttcgat gtattcaaaa tatgcctgaa acactaccca ataattcctg ctattcagct 2820
ggcattgcaa aactggaaga aggagatgaa ctccaacttg caataccaag agaaaatgca 2880
caaatatcac tggatggaga tgtcacattt tttggtgcat tgaaactgct gtga 2934
<210> 18
<211> 786
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VL-light-full-pCR3
<400> 18
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagagc 120
gtgctgaccc agcctcctag cgtgtccgaa gcccctagac agaccgtgac catcagctgc 180
tccggcaaca gcttcaacat cggcagatac cccgtgaact ggtatcagca gctgccaggc 240
aaggccccta aactgctgat ctactataac aacctgcggt tcagcggagt gtccgaccgg 300
ttctctggca gcaagtctgg cacatctgcc agcctggcca tccgggatct gctgtctgag 360
gacgaggccg actactactg cagcacctgg gacgacaccc tgaagggctg ggtgttcggc 420
ggaggcacca aagtgacagt gctgggcgga tccgaaatca agcgtacggt ggccgctccc 480
agcgtgttca tcttcccacc tagcgacgag cagctgaagt ccggcacagc ctctgtcgtg 540
tgcctgctga acaacttcta cccccgcgag gccaaggtgc agtggaaggt ggacaatgcc 600
ctgcagagcg gcaacagcca ggaaagcgtg accgagcagg acagcaagga ctccacctac 660
agcctgagca gcaccctgac cctgagcaag gccgactacg agaagcacaa ggtgtacgcc 720
tgcgaagtga cccaccaggg cctgtctagc cccgtgacca agagcttcaa ccggggcgag 780
tgctaa 786
<210> 19
<211> 963
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 1表达的蛋白质构建体
<400> 19
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Glu Phe Glu Phe Thr Arg Asp Lys Pro Val Ala His
485 490 495
Val Val Ala Asn His Gln Val Glu Glu Gln Leu Glu Trp Leu Ser Gln
500 505 510
Arg Ala Asn Ala Leu Leu Ala Asn Gly Met Asp Leu Lys Asp Asn Gln
515 520 525
Leu Val Val Pro Ala Asp Gly Leu Tyr Leu Val Tyr Ser Gln Val Leu
530 535 540
Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val Leu Leu Thr His Thr Val
545 550 555 560
Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys Val Asn Leu Leu Ser Ala
565 570 575
Val Lys Ser Pro Cys Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys
580 585 590
Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys
595 600 605
Gly Asp Gln Leu Ser Ala Glu Val Asn Leu Pro Lys Tyr Leu Asn Phe
610 615 620
Arg Glu Ser Gly Gln Val Tyr Phe Gly Val Ile Ala Leu Gly Gly Gly
625 630 635 640
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Asp Lys Pro
645 650 655
Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln Leu Glu Trp
660 665 670
Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met Asp Leu Lys
675 680 685
Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu Val Tyr Ser
690 695 700
Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val Leu Leu Thr
705 710 715 720
His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys Val Asn Leu
725 730 735
Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro Glu Gly Ala
740 745 750
Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly Val Phe Gln
755 760 765
Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu Pro Lys Tyr
770 775 780
Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val Ile Ala Leu
785 790 795 800
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
805 810 815
Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln
820 825 830
Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met
835 840 845
Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu
850 855 860
Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val
865 870 875 880
Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys
885 890 895
Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro
900 905 910
Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly
915 920 925
Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu
930 935 940
Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val
945 950 955 960
Ile Ala Leu
<210> 20
<211> 484
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 2表达的蛋白质构建体
<400> 20
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys
<210> 21
<211> 729
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 3表达的蛋白质构建体
<400> 21
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln Pro Gly Ala Glu
485 490 495
Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly
500 505 510
Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly
515 520 525
Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr
530 535 540
Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys
545 550 555 560
Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp
565 570 575
Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp
580 585 590
Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly
595 600 605
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile
610 615 620
Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys
625 630 635 640
Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp
645 650 655
Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr
660 665 670
Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser
675 680 685
Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala
690 695 700
Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly
705 710 715 720
Gly Gly Thr Lys Leu Glu Ile Lys Arg
725
<210> 22
<211> 957
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 4表达的蛋白质构建体
<400> 22
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Leu Glu Gly Pro Glu Glu Thr Val Thr Gln Asp Cys
485 490 495
Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser
500 505 510
Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu
515 520 525
Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe
530 535 540
Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His
545 550 555 560
Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu
565 570 575
Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn
580 585 590
Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu
595 600 605
Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly
610 615 620
Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser
625 630 635 640
Gly Gly Gly Gly Ser Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu
645 650 655
Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr
660 665 670
Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu
675 680 685
Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile
690 695 700
Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu
705 710 715 720
Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val
725 730 735
Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn
740 745 750
Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu
755 760 765
Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp
770 775 780
Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly
785 790 795 800
Gly Gly Gly Ser Gln Phe Ala Ala Gly Pro Glu Glu Thr Val Thr Gln
805 810 815
Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys
820 825 830
Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser
835 840 845
Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr
850 855 860
Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met
865 870 875 880
Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu
885 890 895
Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu
900 905 910
Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly
915 920 925
Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu
930 935 940
Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu
945 950 955
<210> 23
<211> 262
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 5表达的蛋白质构建体
<400> 23
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Val Met Thr Gln Asn Pro Leu Ser
35 40 45
Leu Pro Val Ser Leu Gly Asp Glu Ala Ser Ile Ser Cys Arg Ser Ser
50 55 60
Gln Ser Leu Glu Asn Ser Asn Gly Asn Thr Phe Leu Asn Trp Phe Phe
65 70 75 80
Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Val Ser Asn
85 90 95
Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr
100 105 110
Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val
115 120 125
Tyr Phe Cys Leu Gln Val Thr His Val Pro Tyr Thr Phe Gly Gly Gly
130 135 140
Thr Thr Leu Glu Ile Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala
145 150 155 160
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
165 170 175
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
180 185 190
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
195 200 205
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
210 215 220
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
225 230 235 240
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
245 250 255
Phe Asn Arg Gly Glu Cys
260
<210> 24
<211> 635
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 6表达的蛋白质构建体
<400> 24
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Leu Pro Gly Thr Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser
485 490 495
Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln
500 505 510
His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser Arg Met
515 520 525
Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe Lys Phe
530 535 540
Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu
545 550 555 560
Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln Ser Lys
565 570 575
Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile Arg Val
580 585 590
Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe
595 600 605
Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp Ile Ala
610 615 620
Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln
625 630 635
<210> 25
<211> 732
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 7表达的蛋白质构建体
<400> 25
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val
485 490 495
Ser Leu Gly Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile
500 505 510
Ser Ser Asn Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys
515 520 525
Gly Leu Ile Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg
530 535 540
Phe Ser Gly Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser
545 550 555 560
Leu Glu Ser Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln
565 570 575
Phe Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly
580 585 590
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly
595 600 605
Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys
610 615 620
Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe
625 630 635 640
Ser Ser Ser Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu
645 650 655
Glu Trp Ile Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn
660 665 670
Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
675 680 685
Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val
690 695 700
Tyr Phe Cys Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp
705 710 715 720
Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser
725 730
<210> 26
<211> 726
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 8表达的蛋白质构建体
<400> 26
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys
485 490 495
Pro Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe
500 505 510
Thr Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu
515 520 525
Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn
530 535 540
Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
545 550 555 560
Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val
565 570 575
Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn
580 585 590
Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly
595 600 605
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser
610 615 620
Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met
625 630 635 640
Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln
645 650 655
Lys Pro Gly Pro Ser Ser Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu
660 665 670
Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser
675 680 685
Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr
690 695 700
Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr
705 710 715 720
Lys Leu Glu Ile Lys Arg
725
<210> 27
<211> 481
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 9表达的蛋白质构建体
<400> 27
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys
<210> 28
<211> 907
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 10表达的蛋白质构建体
<400> 28
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gly Gly Gly Ser Gly Gln Leu Glu Thr Ala Lys Glu Pro
485 490 495
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
500 505 510
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
515 520 525
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
530 535 540
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
545 550 555 560
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
565 570 575
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
580 585 590
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
595 600 605
Leu Ala Asn Pro Gln Phe Ile Ser Gly Gly Gly Ser Gly Gly Gly Ser
610 615 620
Gly Gly Gly Ser Gly Gly Gly Ser Gln Leu Glu Thr Ala Lys Glu Pro
625 630 635 640
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
645 650 655
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
660 665 670
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
675 680 685
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
690 695 700
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
705 710 715 720
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
725 730 735
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
740 745 750
Leu Ala Asn Pro Gln Phe Ile Ser Gly Gly Gly Gly Ser Gly Gly Gly
755 760 765
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gln Phe Gln Leu Glu Thr Ala
770 775 780
Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln
785 790 795 800
Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys
805 810 815
Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala
820 825 830
Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr
835 840 845
Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln
850 855 860
Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu
865 870 875 880
Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly
885 890 895
Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
900 905
<210> 29
<211> 1136
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 11表达的蛋白质构建体
<400> 29
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gly Gly Gly Ser Gly Thr Glu Pro Arg Pro Ala Leu Thr
485 490 495
Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln
500 505 510
Val Thr Pro Val Ser His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly
515 520 525
Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys
530 535 540
Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr
545 550 555 560
Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val
565 570 575
Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro
580 585 590
Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val
595 600 605
Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr
610 615 620
Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser
625 630 635 640
Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val Gly
645 650 655
Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp Trp Glu
660 665 670
Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu Val Lys Pro
675 680 685
Asp Asn Pro Trp Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
690 695 700
Ser Gly Gly Gly Ser Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr
705 710 715 720
Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro
725 730 735
Val Ser His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val
740 745 750
Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr
755 760 765
Leu Asn Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln
770 775 780
Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro
785 790 795 800
Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr
805 810 815
Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val Leu Gln Ala
820 825 830
Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr Val Glu Leu
835 840 845
Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser Trp Ser Gln
850 855 860
Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val Gly Leu Arg Ala
865 870 875 880
Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp Trp Glu Leu Ser Tyr
885 890 895
Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu Val Lys Pro Asp Asn Pro
900 905 910
Trp Glu Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly
915 920 925
Gly Gly Ser Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro
930 935 940
Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro Val Ser
945 950 955 960
His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val Phe Ala
965 970 975
Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn
980 985 990
Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu
995 1000 1005
Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro Gly
1010 1015 1020
Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr
1025 1030 1035
Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val Leu Gln
1040 1045 1050
Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr Val
1055 1060 1065
Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser
1070 1075 1080
Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val
1085 1090 1095
Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp
1100 1105 1110
Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu
1115 1120 1125
Val Lys Pro Asp Asn Pro Trp Glu
1130 1135
<210> 30
<211> 611
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 12表达的蛋白质构建体
<400> 30
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Pro Thr Ala Ile Glu Ser Cys Met Val Lys Phe Glu Leu
485 490 495
Ser Ser Ser Lys Trp His Met Thr Ser Pro Lys Pro His Cys Val Asn
500 505 510
Thr Thr Ser Asp Gly Lys Leu Lys Ile Leu Gln Ser Gly Thr Tyr Leu
515 520 525
Ile Tyr Gly Gln Val Ile Pro Val Asp Lys Lys Tyr Ile Lys Asp Asn
530 535 540
Ala Pro Phe Val Val Gln Ile Tyr Lys Lys Asn Asp Val Leu Gln Thr
545 550 555 560
Leu Met Asn Asp Phe Gln Ile Leu Pro Ile Gly Gly Val Tyr Glu Leu
565 570 575
His Ala Gly Asp Asn Ile Tyr Leu Lys Phe Asn Ser Lys Asp His Ile
580 585 590
Gln Lys Thr Asn Thr Tyr Trp Gly Ile Ile Leu Met Pro Asp Leu Pro
595 600 605
Phe Ile Ser
610
<210> 31
<211> 244
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 13表达的蛋白质构建体
<400> 31
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Ile Val Met
20 25 30
Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser
35 40 45
Ile Thr Cys Lys Ala Ser Gln Asp Val Asp Thr Ala Val Ala Trp Tyr
50 55 60
Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser
65 70 75 80
Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly
85 90 95
Thr Asp Tyr Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
100 105 110
Arg Tyr Tyr Cys Gln Gln Tyr Tyr Ser Val Pro Pro Thr Phe Gly Gly
115 120 125
Gly Thr Lys Leu Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
130 135 140
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
145 150 155 160
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
165 170 175
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
180 185 190
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
195 200 205
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
210 215 220
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
225 230 235 240
Arg Gly Glu Cys
<210> 32
<211> 755
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 14表达的蛋白质构建体
<400> 32
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Asp Ile Gln Met Thr Gln
500 505 510
Ser Pro Ser Ser Met Ser Val Ser Leu Gly Asp Thr Val Ser Ile Thr
515 520 525
Cys His Ala Ser Gln Gly Ile Ser Ser Asn Ile Gly Trp Leu Gln Gln
530 535 540
Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile Tyr His Gly Thr Asn Leu
545 550 555 560
Glu Asp Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp
565 570 575
Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser Glu Asp Phe Ala Asp Tyr
580 585 590
Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr Thr Phe Gly Gly Gly Thr
595 600 605
Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
610 615 620
Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser Gln Val Gln Leu Gln Gln
625 630 635 640
Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys
645 650 655
Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser Trp Met Asp Trp Val Lys
660 665 670
Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Arg Ile Tyr Pro Gly
675 680 685
Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu
690 695 700
Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu
705 710 715 720
Thr Ser Val Asp Ser Ala Val Tyr Phe Cys Ala Arg Ser Ile Thr Thr
725 730 735
Val Val Gly Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr
740 745 750
Val Ser Ser
755
<210> 33
<211> 504
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 15表达的蛋白质构建体
<400> 33
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys
500
<210> 34
<211> 533
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 16表达的蛋白质构建体
<400> 34
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Leu Glu Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro
290 295 300
Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
305 310 315 320
Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu
325 330 335
Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln
340 345 350
Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr
355 360 365
Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr
370 375 380
Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val
385 390 395 400
Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser
405 410 415
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln
420 425 430
Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr
435 440 445
Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys
450 455 460
Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala
465 470 475 480
Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr
485 490 495
Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr
500 505 510
Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys
515 520 525
Leu Glu Ile Lys Arg
530
<210> 35
<211> 977
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 17表达的蛋白质构建体
<400> 35
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Gly Pro Glu Glu Thr Val
500 505 510
Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile
515 520 525
Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg
530 535 540
Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr
545 550 555 560
Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr
565 570 575
Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp
580 585 590
Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu
595 600 605
Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu
610 615 620
Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile
625 630 635 640
Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Pro Glu Glu Thr Val Thr
660 665 670
Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln
675 680 685
Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly
690 695 700
Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly
705 710 715 720
Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala
725 730 735
Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu
740 745 750
Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr
755 760 765
Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu
770 775 780
Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser
785 790 795 800
Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly
805 810 815
Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe Ala Ala Gly Pro Glu Glu
820 825 830
Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro
835 840 845
Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe
850 855 860
Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys
865 870 875 880
Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys
885 890 895
Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe
900 905 910
Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met
915 920 925
Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys
930 935 940
Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala
945 950 955 960
Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu
965 970 975
Leu
<210> 36
<211> 261
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 18表达的蛋白质构建体
<400> 36
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Ser Val Leu Thr Gln Pro Pro Ser Val
35 40 45
Ser Glu Ala Pro Arg Gln Thr Val Thr Ile Ser Cys Ser Gly Asn Ser
50 55 60
Phe Asn Ile Gly Arg Tyr Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly
65 70 75 80
Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Asn Asn Leu Arg Phe Ser Gly
85 90 95
Val Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu
100 105 110
Ala Ile Arg Asp Leu Leu Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser
115 120 125
Thr Trp Asp Asp Thr Leu Lys Gly Trp Val Phe Gly Gly Gly Thr Lys
130 135 140
Val Thr Val Leu Gly Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro
145 150 155 160
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
165 170 175
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
180 185 190
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
195 200 205
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
210 215 220
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
225 230 235 240
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
245 250 255
Asn Arg Gly Glu Cys
260
<210> 37
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR1-H
<400> 37
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn
1 5 10
<210> 38
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR2-H
<400> 38
Tyr Ile Arg Tyr Asp Gly Thr Ser Glu Tyr Thr Pro Ser Leu Lys Asn
1 5 10 15
<210> 39
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR1-L
<400> 39
Ser Ser Gln Ser Leu Glu Asn Ser Asn Gly Asn Thr Phe Leu Asn
1 5 10 15
<210> 40
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR2-L
<400> 40
Arg Val Ser Asn Arg Phe Ser
1 5
<210> 41
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5的CDR3-L
<400> 41
Leu Gln Val Thr His Val Pro Tyr Thr
1 5
<210> 42
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR1-H
<400> 42
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val
1 5 10
<210> 43
<211> 21
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR2-H
<400> 43
Ser Ile Ala Tyr Arg Gly Asn Ser Asn Ser Gly Ser Thr Tyr Tyr Asn
1 5 10 15
Pro Ser Leu Lys Ser
20
<210> 44
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR3-H
<400> 44
Arg Gln Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp
1 5 10 15
Val
<210> 45
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR1-L
<400> 45
Ser Gly Asn Ser Phe Asn Ile Gly Arg Tyr Pro Val Asn
1 5 10
<210> 46
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR2-L
<400> 46
Tyr Asn Asn Leu Arg Phe Ser
1 5
<210> 47
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR3-L
<400> 47
Ser Thr Trp Asp Asp Thr Leu Lys Gly Trp Val
1 5 10
<210> 48
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR1-H
<400> 48
Gly Ser Ile Ser Ser Gly Asp Tyr Phe Trp Ser
1 5 10
<210> 49
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR2-H
<400> 49
His Ile His Asn Ser Gly Thr Thr Tyr Tyr Asn Pro Ser Leu Lys Ser
1 5 10 15
<210> 50
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR3-H
<400> 50
Asp Arg Gly Gly Asp Tyr Tyr Tyr Gly Met Asp Val
1 5 10
<210> 51
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR1-L
<400> 51
Arg Ala Ser Gln Gly Ile Ser Arg Ser Tyr Leu Ala
1 5 10
<210> 52
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR2-L
<400> 52
Gly Ala Ser Ser Arg Ala Thr
1 5
<210> 53
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR3-L
<400> 53
Gln Gln Phe Gly Ser Ser Pro Trp Thr
1 5
<210> 54
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR1-H
<400> 54
Tyr Thr Phe Thr Asp Tyr Gly Met His
1 5
<210> 55
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR2-H
<400> 55
Val Ile Ser Thr Tyr Asn Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 56
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR3-H
<400> 56
Ala Tyr Tyr Gly Asn Leu Tyr Tyr Ala Met Asp Tyr
1 5 10
<210> 57
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR1-L
<400> 57
Arg Ala Ser Lys Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His
1 5 10 15
<210> 58
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR2-L
<400> 58
Tyr Ala Ser Asn Leu Glu Ser
1 5
<210> 59
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR3-L
<400> 59
Gln His Ser Arg Glu Leu Pro Phe Thr
1 5
<210> 60
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR1-H
<400> 60
Tyr Ser Phe Thr Ser Tyr Asp Ile Asn
1 5
<210> 61
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR2-H
<400> 61
Trp Ile Tyr Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys
1 5 10 15
Gly
<210> 62
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR3-H
<400> 62
Leu Thr Gly Pro Tyr Trp Tyr Phe Asp Val
1 5 10
<210> 63
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR1-L
<400> 63
Lys Ala Ser Gln Asp Val Asp Thr Ala Val Ala
1 5 10
<210> 64
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR2-L
<400> 64
Trp Ala Ser Thr Arg His Thr
1 5
<210> 65
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR3-L
<400> 65
Gln Gln Tyr Tyr Ser Val Pro Pro Thr
1 5
<210> 66
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR1-H
<400> 66
Tyr Thr Phe Thr Ser Tyr Asn Met His
1 5
<210> 67
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR2-H
<400> 67
Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 68
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR3-H
<400> 68
Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val
1 5 10
<210> 69
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR1-L
<400> 69
Arg Ala Ser Ser Ser Val Ser Tyr Ile His
1 5 10
<210> 70
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR2-L
<400> 70
Ala Thr Ser Asn Leu Ala Ser
1 5
<210> 71
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR3-L
<400> 71
Gln Gln Trp Thr Ser Asn Pro Pro Thr
1 5
<210> 72
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR1-H
<400> 72
Tyr Ala Phe Ser Ser Ser Trp Met Asp
1 5
<210> 73
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR2-H
<400> 73
Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys
1 5 10 15
Gly
<210> 74
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR3-H
<400> 74
Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val
1 5 10
<210> 75
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR1-L
<400> 75
His Ala Ser Gln Gly Ile Ser Ser Asn Ile Gly
1 5 10
<210> 76
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR2-L
<400> 76
His Gly Thr Asn Leu Glu Asp
1 5
<210> 77
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR3-L
<400> 77
Val Gln Tyr Ala Gln Phe Pro Tyr Thr
1 5
<210> 78
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR1-H
<400> 78
Tyr Thr Phe Thr Asn Tyr Gly Met Asn
1 5
<210> 79
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR2-H
<400> 79
Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe Lys
1 5 10 15
Gly
<210> 80
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR3-H
<400> 80
Asp Tyr Gly Asp Tyr Gly Met Asp Tyr
1 5
<210> 81
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR1-L
<400> 81
Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His
1 5 10 15
<210> 82
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR2-L
<400> 82
Leu Ala Ser Asn Leu Glu Ser
1 5
<210> 83
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR3-L
<400> 83
Gln His Ser Arg Glu Val Pro Trp Thr
1 5
<210> 84
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR1-H
<400> 84
Phe Thr Phe Ser Ser Tyr Trp Met Ser
1 5
<210> 85
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR2-H
<400> 85
Glu Ile Arg Leu Lys Ser Asp Asn Tyr Ala Thr His Tyr Ala Glu Ser
1 5 10 15
Val Lys Gly
<210> 86
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR3-H
<400> 86
Gly Tyr Tyr Ala Asp Ala Met Asp Tyr
1 5
<210> 87
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR1-L
<400> 87
Arg Ala Ser Gln Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His
1 5 10 15
<210> 88
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR2-L
<400> 88
Tyr Ala Ser Asn Leu Glu Ser
1 5
<210> 89
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR3-L
<400> 89
Gln His Ser Trp Glu Ile Pro Tyr Thr
1 5
<210> 90
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR1-H
<400> 90
Phe Thr Phe Ser Ser Tyr Ile Met His
1 5
<210> 91
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR2-H
<400> 91
Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 92
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR3-H
<400> 92
Asp Thr Asp Gly Tyr Asp Phe Asp Tyr
1 5
<210> 93
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR1-L
<400> 93
Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
1 5 10
<210> 94
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR2-L
<400> 94
Asp Ala Ser Asn Arg Ala Thr
1 5
<210> 95
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR3-L
<400> 95
Gln Gln Arg Thr Asn Trp Pro Leu Thr
1 5
<210> 96
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 21所示蛋白质构建体的scFv-抗CD20锚定结构域
<400> 96
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 97
<211> 149
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 24所示的蛋白质构建体的鼠源 IL-2 锚定结构域
<400> 97
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln
145
<210> 98
<211> 249
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 25所示的蛋白质构建体的scFv-抗CD19锚定结构域
<400> 98
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val Ser Leu Gly
1 5 10 15
Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile Ser Ser Asn
20 25 30
Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile
35 40 45
Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser
65 70 75 80
Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser
115 120 125
Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
130 135 140
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser
145 150 155 160
Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
165 170 175
Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe
180 185 190
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
195 200 205
Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val Tyr Phe Cys
210 215 220
Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val Trp Gly
225 230 235 240
Ala Gly Thr Thr Val Thr Val Ser Ser
245
<210> 99
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 26所示的蛋白质构建体的scFv-抗CD20锚定结构域
<400> 99
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Pro Ser Ser Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 100
<211> 249
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 32所示的蛋白质构建体的scFv-抗CD19锚定结构域
<400> 100
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val Ser Leu Gly
1 5 10 15
Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile Ser Ser Asn
20 25 30
Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile
35 40 45
Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser
65 70 75 80
Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser
115 120 125
Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
130 135 140
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser
145 150 155 160
Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
165 170 175
Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe
180 185 190
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
195 200 205
Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val Tyr Phe Cys
210 215 220
Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val Trp Gly
225 230 235 240
Ala Gly Thr Thr Val Thr Val Ser Ser
245
<210> 101
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 34所示的蛋白质构建体的scFv-抗CD20锚定结构域
<400> 101
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 102
<211> 473
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 19所示的蛋白质构建体的scTNF80（鼠）锚定结构域
<400> 102
Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln
1 5 10 15
Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met
20 25 30
Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu
35 40 45
Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val
50 55 60
Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys
65 70 75 80
Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro
85 90 95
Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly
100 105 110
Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu
115 120 125
Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val
130 135 140
Ile Ala Leu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly
145 150 155 160
Gly Gly Ser Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val
165 170 175
Glu Glu Gln Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala
180 185 190
Asn Gly Met Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly
195 200 205
Leu Tyr Leu Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro
210 215 220
Asp Tyr Val Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr
225 230 235 240
Gln Glu Lys Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys
245 250 255
Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr
260 265 270
Leu Gly Gly Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu
275 280 285
Val Asn Leu Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr
290 295 300
Phe Gly Val Ile Ala Leu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Ser Gly Gly Gly Ser Asp Lys Pro Val Ala His Val Val Ala Asn
325 330 335
His Gln Val Glu Glu Gln Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala
340 345 350
Leu Leu Ala Asn Gly Met Asp Leu Lys Asp Asn Gln Leu Val Val Pro
355 360 365
Ala Asp Gly Leu Tyr Leu Val Tyr Ser Gln Val Leu Phe Lys Gly Gln
370 375 380
Gly Cys Pro Asp Tyr Val Leu Leu Thr His Thr Val Ser Arg Phe Ala
385 390 395 400
Ile Ser Tyr Gln Glu Lys Val Asn Leu Leu Ser Ala Val Lys Ser Pro
405 410 415
Cys Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu
420 425 430
Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys Gly Asp Gln Leu
435 440 445
Ser Ala Glu Val Asn Leu Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly
450 455 460
Gln Val Tyr Phe Gly Val Ile Ala Leu
465 470
<210> 103
<211> 471
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 22所示的蛋白质构建体的scBaff锚定结构域
<400> 103
Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp
1 5 10 15
Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp
20 25 30
Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys
35 40 45
Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu
50 55 60
Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys
65 70 75 80
Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys
85 90 95
Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala
100 105 110
Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro
115 120 125
Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly
130 135 140
Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
145 150 155 160
Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser
165 170 175
Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu
180 185 190
Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile
195 200 205
Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr
210 215 220
Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val
225 230 235 240
His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile
245 250 255
Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly
260 265 270
Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg
275 280 285
Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala
290 295 300
Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe
305 310 315 320
Ala Ala Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile
325 330 335
Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val
340 345 350
Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu
355 360 365
Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln
370 375 380
Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg
385 390 395 400
Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe
405 410 415
Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr
420 425 430
Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala
435 440 445
Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe
450 455 460
Phe Gly Ala Leu Lys Leu Leu
465 470
<210> 104
<211> 419
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 28所示的蛋白质构建体的scGITRL锚定结构域
<400> 104
Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu
1 5 10 15
Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys
20 25 30
Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile
35 40 45
Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe
50 55 60
Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn
65 70 75 80
Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly
85 90 95
Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn
100 105 110
Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
115 120 125
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
130 135 140
Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu
145 150 155 160
Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys
165 170 175
Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile
180 185 190
Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe
195 200 205
Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn
210 215 220
Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly
225 230 235 240
Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn
245 250 255
Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
260 265 270
Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
275 280 285
Ser Gln Phe Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe
290 295 300
Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys
305 310 315 320
Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu
325 330 335
Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val
340 345 350
Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr
355 360 365
Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu
370 375 380
His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val
385 390 395 400
Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln
405 410 415
Phe Ile Ser
<210> 105
<211> 648
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 29所示的蛋白质构建体的鼠sc41BBL锚定结构域
<400> 105
Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly
1 5 10 15
Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro Val Ser His Ile Gly
20 25 30
Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu
35 40 45
Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser
50 55 60
Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu
65 70 75 80
Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val
85 90 95
Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys
100 105 110
Val Gln Gly Trp Val Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp
115 120 125
Asp Phe Asp Asn Leu Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met
130 135 140
Glu Asn Lys Leu Val Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys
145 150 155 160
Ala Gly His Arg Leu Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala
165 170 175
Gln Asp Ala Tyr Arg Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser
180 185 190
Phe Gly Leu Phe Leu Val Lys Pro Asp Asn Pro Trp Glu Gly Gly Gly
195 200 205
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Glu Pro
210 215 220
Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu
225 230 235 240
Asn Asn Ala Asp Gln Val Thr Pro Val Ser His Ile Gly Cys Pro Asn
245 250 255
Thr Thr Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn
260 265 270
Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly
275 280 285
Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys
290 295 300
Lys Glu Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu
305 310 315 320
Leu Lys Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly
325 330 335
Trp Val Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp
340 345 350
Asn Leu Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys
355 360 365
Leu Val Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His
370 375 380
Arg Leu Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala
385 390 395 400
Tyr Arg Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu
405 410 415
Phe Leu Val Lys Pro Asp Asn Pro Trp Glu Gly Gly Gly Gly Ser Gly
420 425 430
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Glu Pro Arg Pro
435 440 445
Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn
450 455 460
Ala Asp Gln Val Thr Pro Val Ser His Ile Gly Cys Pro Asn Thr Thr
465 470 475 480
Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala
485 490 495
Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly Ala Gly
500 505 510
Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu
515 520 525
Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys
530 535 540
Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly Trp Val
545 550 555 560
Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu
565 570 575
Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val
580 585 590
Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu
595 600 605
Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg
610 615 620
Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu
625 630 635 640
Val Lys Pro Asp Asn Pro Trp Glu
645
<210> 106
<211> 128
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 30所示的蛋白质构建体的鼠GITRL锚定结构域
<400> 106
Pro Thr Ala Ile Glu Ser Cys Met Val Lys Phe Glu Leu Ser Ser Ser
1 5 10 15
Lys Trp His Met Thr Ser Pro Lys Pro His Cys Val Asn Thr Thr Ser
20 25 30
Asp Gly Lys Leu Lys Ile Leu Gln Ser Gly Thr Tyr Leu Ile Tyr Gly
35 40 45
Gln Val Ile Pro Val Asp Lys Lys Tyr Ile Lys Asp Asn Ala Pro Phe
50 55 60
Val Val Gln Ile Tyr Lys Lys Asn Asp Val Leu Gln Thr Leu Met Asn
65 70 75 80
Asp Phe Gln Ile Leu Pro Ile Gly Gly Val Tyr Glu Leu His Ala Gly
85 90 95
Asp Asn Ile Tyr Leu Lys Phe Asn Ser Lys Asp His Ile Gln Lys Thr
100 105 110
Asn Thr Tyr Trp Gly Ile Ile Leu Met Pro Asp Leu Pro Phe Ile Ser
115 120 125
<210> 107
<211> 111
<212> PRT
<213> Artificial Sequence
<220>
<223> 人IL-2锚定结构域
<400> 107
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Glu His Lys Pro Ser Ser Gln Arg Lys Glu Glu Ser Thr Cys
100 105 110
<210> 108
<211> 807
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(G28.5)-VH(1-114)
<400> 108
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcctaa 807
<210> 109
<211> 2226
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(G28.5)-VH(1-114)-scBaff
<400> 109
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgccctcgag ggacccgagg aaactgtgac tcaggactgt 840
ctccagctca ttgccgatag tgaaacccct accatccaga aaggctctta caccttcgtg 900
ccatggctgc tgtcattcaa acggggatct gctctggagg agaaggaaaa caaaatcctg 960
gtcaaggaaa ccggctactt cttcatctac ggccaggtcc tctacaccga caaaacatac 1020
gctatggggc atctcattca gcggaaaaaa gtccacgtgt tcggcgacga actctctctc 1080
gtgacactgt tccggtgtat tcagaacatg cccgagactc tgcccaataa tagctgctac 1140
tctgctggca ttgccaaact ggaggagggc gacgaactcc agctggctat tcctagggaa 1200
aatgcccaga ttagcctgga cggggatgtg acattttttg gcgccctgaa actgctggga 1260
ggcggaggga gtggcggggg aggctctgga cctgaggaaa ctgtgaccca ggattgtctc 1320
cagctcattg ccgatagtga gactcctacc attcagaagg gatcttacac ctttgtgcct 1380
tggctgctgt ctttcaaacg gggctctgct ctggaggaaa aggagaacaa aattctggtc 1440
aaagagactg gctacttctt catctacggc caggtgctgt acaccgacaa aacatacgcc 1500
atgggccatc tcattcagcg gaaaaaagtc cacgtgttcg gcgacgaact ctctctcgtg 1560
acactgttcc ggtgtatcca gaacatgccc gagacactgc ccaataatag ctgctactct 1620
gccggcattg ctaaactgga ggagggggac gaactccagc tggctattcc tagggaaaat 1680
gcccagattt ctctcgatgg ggatgtgaca ttcttcgggg ccctcaaact gctgggaggc 1740
ggcggatctg gcggaggcgg gagtcaattc gcagcaggtc cagaagaaac agtcactcaa 1800
gactgcttgc aactgattgc agacagtgaa acaccaacta tacaaaaagg atcttacaca 1860
tttgttccat ggcttctcag ctttaaaagg ggaagtgccc tagaagaaaa agagaataaa 1920
atattggtca aagaaactgg ttactttttt atatatggtc aggttttata tactgataag 1980
acctacgcca tgggacatct aattcagagg aagaaggtcc atgtctttgg ggatgaattg 2040
agtctggtga ctttgtttcg atgtattcaa aatatgcctg aaacactacc caataattcc 2100
tgctattcag ctggcattgc aaaactggaa gaaggagatg aactccaact tgcaatacca 2160
agagaaaatg cacaaatatc actggatgga gatgtcacat tttttggtgc attgaaactg 2220
ctgtga 2226
<210> 110
<211> 2286
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-VH(1-114)-scBaff
<400> 110
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgccctcgag ggacccgagg aaactgtgac tcaggactgt 900
ctccagctca ttgccgatag tgaaacccct accatccaga aaggctctta caccttcgtg 960
ccatggctgc tgtcattcaa acggggatct gctctggagg agaaggaaaa caaaatcctg 1020
gtcaaggaaa ccggctactt cttcatctac ggccaggtcc tctacaccga caaaacatac 1080
gctatggggc atctcattca gcggaaaaaa gtccacgtgt tcggcgacga actctctctc 1140
gtgacactgt tccggtgtat tcagaacatg cccgagactc tgcccaataa tagctgctac 1200
tctgctggca ttgccaaact ggaggagggc gacgaactcc agctggctat tcctagggaa 1260
aatgcccaga ttagcctgga cggggatgtg acattttttg gcgccctgaa actgctggga 1320
ggcggaggga gtggcggggg aggctctgga cctgaggaaa ctgtgaccca ggattgtctc 1380
cagctcattg ccgatagtga gactcctacc attcagaagg gatcttacac ctttgtgcct 1440
tggctgctgt ctttcaaacg gggctctgct ctggaggaaa aggagaacaa aattctggtc 1500
aaagagactg gctacttctt catctacggc caggtgctgt acaccgacaa aacatacgcc 1560
atgggccatc tcattcagcg gaaaaaagtc cacgtgttcg gcgacgaact ctctctcgtg 1620
acactgttcc ggtgtatcca gaacatgccc gagacactgc ccaataatag ctgctactct 1680
gccggcattg ctaaactgga ggagggggac gaactccagc tggctattcc tagggaaaat 1740
gcccagattt ctctcgatgg ggatgtgaca ttcttcgggg ccctcaaact gctgggaggc 1800
ggcggatctg gcggaggcgg gagtcaattc gcagcaggtc cagaagaaac agtcactcaa 1860
gactgcttgc aactgattgc agacagtgaa acaccaacta tacaaaaagg atcttacaca 1920
tttgttccat ggcttctcag ctttaaaagg ggaagtgccc tagaagaaaa agagaataaa 1980
atattggtca aagaaactgg ttactttttt atatatggtc aggttttata tactgataag 2040
acctacgcca tgggacatct aattcagagg aagaaggtcc atgtctttgg ggatgaattg 2100
agtctggtga ctttgtttcg atgtattcaa aatatgcctg aaacactacc caataattcc 2160
tgctattcag ctggcattgc aaaactggaa gaaggagatg aactccaact tgcaatacca 2220
agagaaaatg cacaaatatc actggatgga gatgtcacat tttttggtgc attgaaactg 2280
ctgtga 2286
<210> 111
<211> 2169
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG2-VH-scFv:CD20
<400> 111
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctcg agtgctagca ccaagggccc atcggtcttc 480
cccctggcgc cctgctccag gagcacctcc gagagcacag cggccctggg ctgcctggtc 540
aaggactact tccccgaacc ggtgacggtg tcgtggaact caggcgctct gaccagcggc 600
gtgcacacct tcccagctgt cctacagtcc tcaggactct actccctcag cagcgtggtg 660
accgtgccct ccagcaactt cggcacccag acctacacct gcaacgtaga tcacaagccc 720
agcaacacca aggtggacaa gacagttgag cgcaaatgtt gtgtcgagtg cccaccgtgc 780
ccagcaccac ctgtggcagg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 840
ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccacgaagac 900
cccgaggtcc agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 960
ccacgggagg agcagttcaa cagcacgttc cgtgtggtca gcgtcctcac cgttgtgcac 1020
caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccagcc 1080
cccatcgaga aaaccatctc caaaaccaaa gggcagcccc gagaaccaca ggtgtacacc 1140
ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa 1200
ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 1260
tacaagacca cgcctcccat gctggactcc gacggctcct tcttcctcta cagcaagctc 1320
accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 1380
gctctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa agaattccag 1440
gtacaactgc agcagcctgg ggctgagctg gtgaagcctg gggcctcagt gaagatgtcc 1500
tgcaaggctt ctggctacac atttaccagt tacaatatgc actgggtaaa acagacacct 1560
ggtcggggcc tggaatggat tggagctatt tatcccggaa atggtgatac ttcctacaat 1620
cagaagttca aaggcaaggc cacattgact gcagacaaat cctccagcac agcctacatg 1680
cagctcagca gcctgacatc tgaggactct gcggtctatt actgtgcaag atcgacttac 1740
tacggcggtg actggtactt caatgtctgg ggcgcaggga ccacggtcac cgtctcttca 1800
ggaggaggcg gatccggcgg aggcggaagc ggtggcggag gctctcaaat tgttctctcc 1860
cagtctccag caatcctgtc tgcatctcca ggggagaagg tcacaatgac ttgcagggcc 1920
agctcaagtg taagttacat ccactggttc cagcagaagc caggatcctc ccccaaaccc 1980
tggatttatg ccacatccaa cctggcttct ggagtccctg ttcgcttcag tggcagtggg 2040
tctgggactt cttactctct cacaatcagc agagtggagg ctgaagatgc tgccacttat 2100
tactgccagc agtggactag taacccaccc acgttcggag gggggaccaa gctggaaatc 2160
aaacgttaa 2169
<210> 112
<211> 2196
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(1F6)
<400> 112
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc agattcagct cgtccagtcc ggacctgaag tgaaaaaacc tggcgaaacc 1500
gtgaaaattt cctgtaaggc ctctggctac acctttacca actacggcat gaactgggtc 1560
aaacaggctc ctgggaaggg cctgaaatgg atgggatgga tcaacaccta caccggcgaa 1620
ccaacatacg ccgatgcctt taagggacgc tttgccttct ctctggaaac ttccgcctct 1680
actgcttacc tccagatcaa taacctcaaa aacgaggaca ccgccactta cttttgtgct 1740
cgggattacg gggactacgg gatggattac tggggacagg gaacatctgt gaccgtgtct 1800
agcgcttcta caaaggggcc taaactggag gagggcgagt ttagcgaggc tagagtggat 1860
atcgtgctca cacagtctcc cgcttctctg gctgtctcac tgggccagcg agcaacaatc 1920
tcttgtcggg cttccaaatc cgtgtctact agcggctact cttttatgca ctggtaccag 1980
cagaaacctg ggcagcctcc aaaactgctc atctacctgg cttcaaacct cgaatccgga 2040
gtgcctgctc gattttctgg ctctggctcc gggaccgact ttacactgaa cattcatcct 2100
gtcgaggagg aggacgctgc cacatactac tgtcagcatt ctagggaggt gccatggaca 2160
tttggcgggg gaacaaaact ggaaatcaaa cggtaa 2196
<210> 113
<211> 2187
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(2H5)
<400> 113
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc aggtgcagct ggtggaatct ggcggcggag tggtgcagcc tggcagaagc 1500
ctgagactga gctgtgccgc cagcggcttc accttcagca gctacatcat gcactgggtg 1560
cgccaggccc ctggcaaggg actggaatgg gtggccgtga tcagctacga cggccggaac 1620
aagtactacg ccgacagcgt gaagggccgg ttcaccatct cccgggacaa cagcaagaac 1680
accctgtacc tgcagatgaa cagcctgcgg gccgaggaca ccgccgtgta ctactgtgcc 1740
agagacaccg acggctacga cttcgactat tggggccagg gcaccctcgt gaccgtgtct 1800
agcggaggcg gaggatctgg cggaggggga tcaggcgggg gaggctctga aatcgtgctg 1860
acacagagcc ccgccaccct gtcactgtct ccaggcgaaa gagccaccct gagctgcaga 1920
gccagccaga gcgtgtccag ctacctggcc tggtatcagc agaagcccgg acaggccccc 1980
agactgctga tctacgacgc cagcaatcgg gccacaggca tccctgccag attttccggc 2040
tctggcagcg gcaccgactt caccctgaca atcagcagcc tggaacccga ggactttgcc 2100
gtgtattatt gccagcagcg gaccaactgg cccctgacct ttggcggagg caccaaggtg 2160
gaaatcaagg ccagcaccaa gggctaa 2187
<210> 114
<211> 2211
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK4)-IgG1(N297A)-VH-scFv:CD20
<400> 114
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatccagc 480
agcgcctcta caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgaccgtg 600
tcctggaact ctggcgctct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg acagtgccca gcagctctct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgcccccga actgctggga 840
ggcccttccg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 900
cccgaagtga cctgcgtggt ggtggatgtg tcccacgagg accctgaagt gaagtttaat 960
tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcctagaga ggaacagtac 1020
gccagcacct accgggtggt gtccgtgctg acagtgctgc accaggactg gctgaacggc 1080
aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 1140
agcaaggcca agggccagcc ccgcgaaccc caggtgtaca cactgccccc aagcagggac 1200
gagctgacca agaaccaggt gtccctgacc tgtctcgtga aaggcttcta ccccagcgat 1260
atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 1320
gtgctggaca gcgacggctc attcttcctg tacagcaagc tgaccgtgga caagtcccgg 1380
tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1440
acccagaagt ccctgagcct gagccccggc aagctcgagc aggtacaact gcagcagcct 1500
ggggctgagc tggtgaagcc tggggcctca gtgaagatgt cctgcaaggc ttctggctac 1560
acatttacca gttacaatat gcactgggta aaacagacac ctggtcgggg cctggaatgg 1620
attggagcta tttatcccgg aaatggtgat acttcctaca atcagaagtt caaaggcaag 1680
gccacattga ctgcagacaa atcctccagc acagcctaca tgcagctcag cagcctgaca 1740
tctgaggact ctgcggtcta ttactgtgca agatcgactt actacggcgg tgactggtac 1800
ttcaatgtct ggggcgcagg gaccacggtc accgtctctt caggaggagg cggatccggc 1860
ggaggcggaa gcggtggcgg aggctctcaa attgttctct cccagtctcc agcaatcctg 1920
tctgcatctc caggggagaa ggtcacaatg acttgcaggg ccagctcaag tgtaagttac 1980
atccactggt tccagcagaa gccaggatcc tcccccaaac cctggattta tgccacatcc 2040
aacctggctt ctggagtccc tgttcgcttc agtggcagtg ggtctgggac ttcttactct 2100
ctcacaatca gcagagtgga ggctgaagat gctgccactt attactgcca gcagtggact 2160
agtaacccac ccacgttcgg aggggggacc aagctggaaa tcaaacgtta a 2211
<210> 115
<211> 774
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK4)-VL
<400> 115
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
gtgatgactc agagccccgc cttcctgtct gtgacccctg gcgagaaagt gaccatcacc 180
tgtagagcca gccagaccat cagcgactac ctgcactggt atcagcagaa gcccgatcag 240
gcccctaagc tgctgattaa gtacgcctct cagagcatca gcggcatccc cagcagattt 300
tctggcagcg gctctggcac cgacttcacc tttaccatca gctccctgga agccgaggat 360
gccgccacct actattgtca ggacggccac agcttccctc caacctttgg acagggcaca 420
aagctggaaa tcaagggatc cgaaatcaag cgtacggtgg ccgctcccag cgtgttcatc 480
ttcccaccta gcgacgagca gctgaagtcc ggcacagcct ctgtcgtgtg cctgctgaac 540
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaatgccct gcagagcggc 600
aacagccagg aaagcgtgac cgagcaggac agcaaggact ccacctacag cctgagcagc 660
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaagtgacc 720
caccagggcc tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaa 774
<210> 116
<211> 2250
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-IgG1(N297A)-VH-scFv:CD20
<220>
<221> misc_feature
<222> (1581)..(1581)
<223> n is a, c, g, or t
<400> 116
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatccagca gcgcctctac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca aggaattcca ggtacaactg cagcagcctg gggctgagct ggtgaagcct 1560
ggggcctcag tgaagatgtc ntgcaaggct tctggctaca catttaccag ttacaatatg 1620
cactgggtaa aacagacacc tggtcggggc ctggaatgga ttggagctat ttatcccgga 1680
aatggtgata cttcctacaa tcagaagttc aaaggcaagg ccacattgac tgcagacaaa 1740
tcctccagca cagcctacat gcagctcagc agcctgacat ctgaggactc tgcggtctat 1800
tactgtgcaa gatcgactta ctacggcggt gactggtact tcaatgtctg gggcgcaggg 1860
accacggtca ccgtctcttc aggaggaggc ggatccggcg gaggcggaag cggtggcgga 1920
ggctctcaaa ttgttctctc ccagtctcca gcaatcctgt ctgcatctcc aggggagaag 1980
gtcacaatga cttgcagggc cagctcaagt gtaagttaca tccactggtt ccagcagaag 2040
ccaggatcct cccccaaacc ctggatttat gccacatcca acctggcttc tggagtccct 2100
gttcgcttca gtggcagtgg gtctgggact tcttactctc tcacaatcag cagagtggag 2160
gctgaagatg ctgccactta ttactgccag cagtggacta gtaacccacc cacgttcgga 2220
ggggggacca agctggaaat caaacgttaa 2250
<210> 117
<211> 268
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 108所示的蛋白质构建体
<400> 117
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
260 265
<210> 118
<211> 741
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 109所示的蛋白质构建体
<400> 118
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Leu Glu Gly Pro
260 265 270
Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu
275 280 285
Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu
290 295 300
Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu
305 310 315 320
Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr
325 330 335
Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His
340 345 350
Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln
355 360 365
Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile
370 375 380
Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu
385 390 395 400
Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu
405 410 415
Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Pro Glu
420 425 430
Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr
435 440 445
Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser
450 455 460
Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val
465 470 475 480
Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp
485 490 495
Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val
500 505 510
Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn
515 520 525
Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala
530 535 540
Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn
545 550 555 560
Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys
565 570 575
Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe Ala Ala
580 585 590
Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp
595 600 605
Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp
610 615 620
Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys
625 630 635 640
Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu
645 650 655
Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys
660 665 670
Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys
675 680 685
Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala
690 695 700
Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro
705 710 715 720
Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly
725 730 735
Ala Leu Lys Leu Leu
740
<210> 119
<211> 761
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 110所示的蛋白质构建体
<400> 119
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Leu Glu Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile
290 295 300
Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val
305 310 315 320
Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu
325 330 335
Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln
340 345 350
Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg
355 360 365
Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe
370 375 380
Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr
385 390 395 400
Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala
405 410 415
Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe
420 425 430
Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly
435 440 445
Ser Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala
450 455 460
Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro
465 470 475 480
Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn
485 490 495
Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val
500 505 510
Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys
515 520 525
Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg
530 535 540
Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser
545 550 555 560
Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile
565 570 575
Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe
580 585 590
Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
595 600 605
Gln Phe Ala Ala Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln
610 615 620
Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr
625 630 635 640
Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu
645 650 655
Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr
660 665 670
Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile
675 680 685
Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr
690 695 700
Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser
705 710 715 720
Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln
725 730 735
Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val
740 745 750
Thr Phe Phe Gly Ala Leu Lys Leu Leu
755 760
<210> 120
<211> 722
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 111所示的蛋白质构建体
<400> 120
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu
245 250 255
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
260 265 270
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
275 280 285
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln
290 295 300
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
305 310 315 320
Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu
325 330 335
Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
340 345 350
Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
355 360 365
Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
370 375 380
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
385 390 395 400
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
405 410 415
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly
420 425 430
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
435 440 445
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
450 455 460
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln
465 470 475 480
Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser
485 490 495
Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn
500 505 510
Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly
515 520 525
Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys
530 535 540
Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met
545 550 555 560
Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala
565 570 575
Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala
580 585 590
Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
595 600 605
Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala
610 615 620
Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala
625 630 635 640
Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser
645 650 655
Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val
660 665 670
Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr
675 680 685
Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
690 695 700
Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile
705 710 715 720
Lys Arg
<210> 121
<211> 731
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 112所示的蛋白质构建体
<400> 121
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Val Lys Lys
485 490 495
Pro Gly Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe
500 505 510
Thr Asn Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu
515 520 525
Lys Trp Met Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala
530 535 540
Asp Ala Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser
545 550 555 560
Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr
565 570 575
Tyr Phe Cys Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly
580 585 590
Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys
595 600 605
Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg Val Asp Ile Val Leu Thr
610 615 620
Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile
625 630 635 640
Ser Cys Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met
645 650 655
His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr
660 665 670
Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
675 680 685
Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu
690 695 700
Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg Glu Val Pro Trp Thr
705 710 715 720
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
725 730
<210> 122
<211> 728
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 113所示的蛋白质构建体
<400> 122
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
485 490 495
Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
500 505 510
Ser Ser Tyr Ile Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
515 520 525
Glu Trp Val Ala Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala
530 535 540
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
545 550 555 560
Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
565 570 575
Tyr Tyr Cys Ala Arg Asp Thr Asp Gly Tyr Asp Phe Asp Tyr Trp Gly
580 585 590
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
595 600 605
Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro
610 615 620
Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
625 630 635 640
Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
645 650 655
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr
660 665 670
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
675 680 685
Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
690 695 700
Gln Gln Arg Thr Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val
705 710 715 720
Glu Ile Lys Ala Ser Thr Lys Gly
725
<210> 123
<211> 736
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 114所示的蛋白质构建体
<400> 123
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
275 280 285
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
290 295 300
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
305 310 315 320
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
325 330 335
Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
340 345 350
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
355 360 365
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
370 375 380
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
385 390 395 400
Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
405 410 415
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
420 425 430
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
435 440 445
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
450 455 460
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
465 470 475 480
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Gln Val Gln
485 490 495
Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys
500 505 510
Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His
515 520 525
Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile
530 535 540
Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys
545 550 555 560
Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu
565 570 575
Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser
580 585 590
Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr
595 600 605
Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
610 615 620
Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu
625 630 635 640
Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser
645 650 655
Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro
660 665 670
Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val
675 680 685
Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser
690 695 700
Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr
705 710 715 720
Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
725 730 735
<210> 124
<211> 257
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 115所示的蛋白质构建体
<400> 124
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Val Met Thr Gln Ser Pro Ala Phe
35 40 45
Leu Ser Val Thr Pro Gly Glu Lys Val Thr Ile Thr Cys Arg Ala Ser
50 55 60
Gln Thr Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Pro Asp Gln
65 70 75 80
Ala Pro Lys Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile
85 90 95
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr
100 105 110
Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asp
115 120 125
Gly His Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
130 135 140
Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
145 150 155 160
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
165 170 175
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
180 185 190
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
195 200 205
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
210 215 220
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
225 230 235 240
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
245 250 255
Cys
<210> 125
<211> 749
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 116所示的蛋白质构建体
<400> 125
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln
500 505 510
Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys
515 520 525
Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys
530 535 540
Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly
545 550 555 560
Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu
565 570 575
Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu
580 585 590
Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr
595 600 605
Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr
610 615 620
Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
625 630 635 640
Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser
645 650 655
Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser
660 665 670
Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp
675 680 685
Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser
690 695 700
Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu
705 710 715 720
Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro
725 730 735
Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
740 745
<210> 126
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR1-H
<400> 126
Tyr Thr Phe Ser Ser Tyr Trp Met His
1 5
<210> 127
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR2-H
<400> 127
Glu Ile Asn Pro Gly Asn Gly His Thr Asn Tyr Asn Glu Lys Phe Lys
1 5 10 15
Ser
<210> 128
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR3-H
<400> 128
Ser Phe Thr Thr Ala Arg Ala Phe Ala Tyr
1 5 10
<210> 129
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR1-L
<400> 129
Arg Ala Ser Gln Thr Ile Ser Asp Tyr Leu His
1 5 10
<210> 130
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR2-L
<400> 130
Leu Ala Ser Gln Ser Ile Ser
1 5
<210> 131
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR3-L
<400> 131
Gln Asp Gly His Ser Phe Pro Pro Thr
1 5
<210> 132
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR1-H
<400> 132
Gly Ser Phe Ser Gly Tyr Tyr Trp Ser
1 5
<210> 133
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR2-H
<400> 133
Glu Ile Asn His Gly Gly Tyr Val Thr Tyr Asn Pro Ser Leu Glu Ser
1 5 10 15
<210> 134
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR3-H
<400> 134
Asp Tyr Gly Pro Gly Asn Tyr Asp Trp Tyr Phe Asp Leu
1 5 10
<210> 135
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR1-L
<400> 135
Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
1 5 10
<210> 136
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR2-L
<400> 136
Asp Ala Ser Asn Arg Ala Thr
1 5
<210> 137
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR3-L
<400> 137
Gln Gln Arg Ser Asn Trp Pro Pro Ala Leu Thr
1 5 10
<210> 138
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR1-H
<400> 138
Tyr Ser Phe Ser Thr Tyr Trp Ile Ser
1 5
<210> 139
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR2-H
<400> 139
Lys Ile Tyr Pro Gly Asp Ser Tyr Thr Asn Tyr Ser Pro Ser Phe Gln
1 5 10 15
Gly
<210> 140
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR3-H
<400> 140
Gly Tyr Gly Ile Phe Asp Tyr
1 5
<210> 141
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR1-L
<400> 141
Ser Gly Asp Asn Ile Gly Asp Gln Tyr Ala His
1 5 10
<210> 142
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR2-L
<400> 142
Gln Asp Lys Asn Arg Pro Ser
1 5
<210> 143
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR3-L
<400> 143
Ala Thr Tyr Thr Gly Phe Gly Ser Leu Ala Val
1 5 10
<210> 144
<211> 248
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 121所示的蛋白质构建体的scFv:CD70(1F6)锚定结构域
<400> 144
Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30
Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met
35 40 45
Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe
50 55 60
Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80
Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys
85 90 95
Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys Leu Glu Glu
115 120 125
Gly Glu Phe Ser Glu Ala Arg Val Asp Ile Val Leu Thr Gln Ser Pro
130 135 140
Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg
145 150 155 160
Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His Trp Tyr
165 170 175
Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser
180 185 190
Asn Leu Glu Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
195 200 205
Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala
210 215 220
Thr Tyr Tyr Cys Gln His Ser Arg Glu Val Pro Trp Thr Phe Gly Gly
225 230 235 240
Gly Thr Lys Leu Glu Ile Lys Arg
245
<210> 145
<211> 245
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 122所示的蛋白质构建体的scFv:CD70(2H5)锚定结构域
<400> 145
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Thr Asp Gly Tyr Asp Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln
145 150 155 160
Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
165 170 175
Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro
180 185 190
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
195 200 205
Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg
210 215 220
Thr Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
225 230 235 240
Ala Ser Thr Lys Gly
245
<210> 146
<211> 2247
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TRAILR2(Cona)-IgG1(N297A)-VH-scFv:CD70(9G2)
<400> 146
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctgcagg aatctggccc tggcctcgtg aagcctagcc agaccctgag cctgacctgt 180
accgtgtctg gcggcagcat cagcagcggc gactacttct ggtcctggat cagacagctg 240
cccggcaagg gcctggaatg gatcggccac atccacaaca gcggcaccac ctactacaac 300
cccagcctga agtccagagt gaccatcagc gtggacacca gcaagaagca gttcagcctg 360
cggctgagca gcgtgacagc cgccgataca gccgtgtact actgcgccag agacagaggc 420
ggcgattact actacggcat ggacgtgtgg ggccagggca ccaccgtgac cgtgtctagc 480
agatccagca gcgcctctac aaagggcccc agcgtgttcc ctctggcccc tagcagcaag 540
agcacatctg gcggaacagc cgccctgggc tgcctcgtga aggactactt tcccgagccc 600
gtgaccgtgt cctggaactc tggcgctctg acaagcggcg tgcacacctt tccagccgtg 660
ctgcagagca gcggcctgta ctctctgagc agcgtcgtga cagtgcccag cagctctctg 720
ggcacccaga cctacatctg caacgtgaac cacaagccca gcaacaccaa ggtggacaag 780
aaggtggaac ccaagagctg cgacaagacc cacacctgtc ccccttgtcc tgcccccgaa 840
ctgctgggag gcccttccgt gttcctgttc cccccaaagc ccaaggacac cctgatgatc 900
agccggaccc ccgaagtgac ctgcgtggtg gtggatgtgt cccacgagga ccctgaagtg 960
aagtttaatt ggtacgtgga cggcgtggaa gtgcacaacg ccaagaccaa gcctagagag 1020
gaacagtacg ccagcaccta ccgggtggtg tccgtgctga cagtgctgca ccaggactgg 1080
ctgaacggca aagagtacaa gtgcaaggtg tccaacaagg ccctgcctgc ccccatcgag 1140
aaaaccatca gcaaggccaa gggccagccc cgcgaacccc aggtgtacac actgccccca 1200
agcagggacg agctgaccaa gaaccaggtg tccctgacct gtctcgtgaa aggcttctac 1260
cccagcgata tcgccgtgga atgggagagc aacggccagc ccgagaacaa ctacaagacc 1320
accccccctg tgctggacag cgacggctca ttcttcctgt acagcaagct gaccgtggac 1380
aagtcccggt ggcagcaggg caacgtgttc agctgcagcg tgatgcacga ggccctgcac 1440
aaccactaca cccagaagtc cctgagcccc ggcaagctcg agcaggtgca gctggtggaa 1500
tctggcggcg gactgatgca gcctggcggc tctctgagac tgagctgtgc cgccagcggc 1560
ttcaccttta gcagcagcgc catgagctgg gtgcgccagg ctcctggaaa gggcctggaa 1620
tgggtgtcca gcatctacag cgacagcagc tacacctact acgccgacag cgtgaagtcc 1680
cggttcacca tcagcaccga caacgccaag aacaccctgt acctgcagat gaacagcctg 1740
aagcccgacg acaccgccgt gtactactgt gccggcagca gcgattacga gggcagcttt 1800
gcctcttggg gccagggcac acaagtgacc gtgtcctcca gatctagcac caagggcccc 1860
aagctggaag agggcgagtt cagcgaggcc caattgcaga gcgtcgtgac ccagcctcct 1920
agcctgtctg cctctcctgg aagcagcgtg cggctgacct gtacactgag cagcggcaac 1980
agcgtgggca actacgacat cagctggtat cagcagaagg ccggcagccc ccccagatac 2040
ctgctgtact actacagcga ttccgtgaag caccagggca gcggcgtgcc cagcagattt 2100
tccggaagct ctgacgccag cgccaacgcc ggactgctgc tgatttctgg cctgcagcct 2160
gaggacgagg ccgactacta ctgcagcgcc tacaagagcg gcagccacgt gttcggcgga 2220
ggcaccaaac tgacagtgct gggctaa 2247
<210> 147
<211> 768
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TRAILR2(Cona)-VL
<400> 147
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggagatc 120
gtgctgaccc agagccctgg caccctgtca ctgtctccag gcgagagagc caccctgagc 180
tgtagagcca gccagggcat cagccggtct tacctggcct ggtatcagca gaagcccggc 240
caggctccta gcctgctgat ctacggcgcc agcagcagag ccaccggcat ccccgataga 300
ttttccggca gcggctccgg caccgacttc accctgacaa tcagcagact ggaacccgag 360
gacttcgccg tgtattattg ccagcagttc ggcagcagcc cctggacctt tggccaggga 420
acaaaagtgg gatccgaaat caagcgtacg gtggccgctc ccagcgtgtt catcttccca 480
cctagcgacg agcagctgaa gtccggcaca gcctctgtcg tgtgcctgct gaacaacttc 540
tacccccgcg aggccaaggt gcagtggaag gtggacaatg ccctgcagag cggcaacagc 600
caggaaagcg tgaccgagca ggacagcaag gactccacct acagcctgag cagcaccctg 660
accctgagca aggccgacta cgagaagcac aaggtgtacg cctgcgaagt gacccaccag 720
ggcctgtcta gccccgtgac caagagcttc aaccggggcg agtgctaa 768
<210> 148
<211> 2226
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
<400> 148
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctggttc agtctggcgc cgaagtgaaa aagcctggcg cctctgtgaa ggtgtcctgt 180
acagccagcg gcttcaacat caaggactac tacgtgcact gggtcaagca ggcccctgga 240
caaggactgg aatggatggg cagaatcgac cccgaggacg gcgactctaa gtacgcccct 300
aagttccagg gcaaagccac catgaccgcc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcaccaccag ctactatgtg 420
ggcacctacg gctattgggg ccagggcaca ctggtcaccg tgtccagcag atccagcagc 480
gcctctacaa agggccccag cgtgttccct ctggccccta gcagcaagag cacatctggc 540
ggaacagccg ccctgggctg cctcgtgaag gactactttc ccgagcccgt gaccgtgtcc 600
tggaactctg gcgctctgac aagcggcgtg cacacctttc cagccgtgct gcagagcagc 660
ggcctgtact ctctgagcag cgtcgtgaca gtgcccagca gctctctggg cacccagacc 720
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggtggaaccc 780
aagagctgcg acaagaccca cacctgtccc ccttgtcctg cccccgaact gctgggaggc 840
ccttccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc 900
gaagtgacct gcgtggtggt ggatgtgtcc cacgaggacc ctgaagtgaa gtttaattgg 960
tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ctagagagga acagtacgcc 1020
agcacctacc gggtggtgtc cgtgctgaca gtgctgcacc aggactggct gaacggcaaa 1080
gagtacaagt gcaaggtgtc caacaaggcc ctgcctgccc ccatcgagaa aaccatcagc 1140
aaggccaagg gccagccccg cgaaccccag gtgtacacac tgcccccaag cagggacgag 1200
ctgaccaaga accaggtgtc cctgacctgt ctcgtgaaag gcttctaccc cagcgatatc 1260
gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg 1320
ctggacagcg acggctcatt cttcctgtac agcaagctga ccgtggacaa gtcccggtgg 1380
cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1440
cagaagtccc tgagcctgag ccccggcaag ctcgaggagg tgcagctgct ggaatctggc 1500
ggaggacttg ttcagcctgg cggctctctg agactgtctt gtgccgccag cggcttcacc 1560
ttcagcagct atatcatgat gtgggtccga caggcccctg gcaaaggcct tgaatgggtg 1620
tccagcatct atcccagcgg cggcatcacc ttttacgccg acacagtgaa gggcagattc 1680
accatcagcc gggacaacag caagaacacc ctgtacctgc agatgaacag cctgagagcc 1740
gaggacaccg ccgtgtacta ctgcgccaga atcaagctgg gcaccgtgac caccgtggat 1800
tattggggac agggcaccct ggtcaccgtg tcctccagat cttctacaaa gggccccaag 1860
ctggaagagg gcgagtttag cgaagcccaa ttgcagagcg ccctgacaca gcctgcatcc 1920
gtgtctggat ctccaggcca gagcatcacc atctcttgta ccggcacaag ctccgatgtc 1980
ggcggctaca attacgtgtc ctggtatcag cagcaccccg gcaaggcccc taagctgatg 2040
atctacgacg tgtccaacag accctccggc gtgtccaata gattcagcgg cagcaagagc 2100
ggcaacaccg ccagcctgac aattagcgga ctgcaggccg aggacgaggc cgattactac 2160
tgtagcagct acaccagctc ctccaccaga gtgtttggca ccggcaccaa agtgaccgtg 2220
ctttaa 2226
<210> 149
<211> 771
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-VL
<400> 149
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
cagatgacac agagccccag cagcctgtct gccagcgtgg gagatagagt gaccatcacc 180
tgtagcgcca gcagcagcgt gtcctacatg ctgtggttcc agcagaagcc tggcaaggcc 240
cctaagctgc tgatctacag cacctccaat ctggccagcg gcgtgccaag cagattttct 300
ggctctggca gcggcaccga cttcaccctg accatatcta gcctgcagcc agaggacttc 360
gccacctact actgccagca gcggacattc tacccctaca cctttggcgg aggcaccaag 420
gtggaaatca agggatccga aatcaagcgt acggtggccg ctcccagcgt gttcatcttc 480
ccacctagcg acgagcagct gaagtccggc acagcctctg tcgtgtgcct gctgaacaac 540
ttctaccccc gcgaggccaa ggtgcagtgg aaggtggaca atgccctgca gagcggcaac 600
agccaggaaa gcgtgaccga gcaggacagc aaggactcca cctacagcct gagcagcacc 660
ctgaccctga gcaaggccga ctacgagaag cacaaggtgt acgcctgcga agtgacccac 720
cagggcctgt ctagccccgt gaccaagagc ttcaaccggg gcgagtgcta a 771
<210> 150
<211> 1578
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-Fab2-scFv:PD-L1(Ave)
<400> 150
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctggttc agtctggcgc cgaagtgaaa aagcctggcg cctctgtgaa ggtgtcctgt 180
acagccagcg gcttcaacat caaggactac tacgtgcact gggtcaagca ggcccctgga 240
caaggactgg aatggatggg cagaatcgac cccgaggacg gcgactctaa gtacgcccct 300
aagttccagg gcaaagccac catgaccgcc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcaccaccag ctactatgtg 420
ggcacctacg gctattgggg ccagggcaca ctggtcaccg tgtccagcag atcctctagc 480
gccagcacaa agggccccag cgtgttccct ctggccccta gcagcaagag cacatctggc 540
ggaacagccg ccctgggctg cctcgtgaag gactactttc ccgagcccgt gacagtgtcc 600
tggaactctg gcgccctgac aagcggcgtg cacacctttc cagccgtgct gcagagcagc 660
ggcctgtact ctctgagcag cgtcgtgact gtgcccagca gcagcctggg cacccagacc 720
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggtggaaccc 780
aagagctgcg acaagaccca cacctgtccc ccttgtcctg ccctcgagga ggtgcagctg 840
ctggaatctg gcggaggact tgttcagcct ggcggctctc tgagactgtc ttgtgccgcc 900
agcggcttca ccttcagcag ctatatcatg atgtgggtcc gacaggcccc tggcaaaggc 960
cttgaatggg tgtccagcat ctatcccagc ggcggcatca ccttttacgc cgacacagtg 1020
aagggcagat tcaccatcag ccgggacaac agcaagaaca ccctgtacct gcagatgaac 1080
agcctgagag ccgaggacac cgccgtgtac tactgcgcca gaatcaagct gggcaccgtg 1140
accaccgtgg attattgggg acagggcacc ctggtcaccg tgtcctccag atcttctaca 1200
aagggcccca agctggaaga gggcgagttt agcgaagccc aattgcagag cgccctgaca 1260
cagcctgcat ccgtgtctgg atctccaggc cagagcatca ccatctcttg taccggcaca 1320
agctccgatg tcggcggcta caattacgtg tcctggtatc agcagcaccc cggcaaggcc 1380
cctaagctga tgatctacga cgtgtccaac agaccctccg gcgtgtccaa tagattcagc 1440
ggcagcaaga gcggcaacac cgccagcctg acaattagcg gactgcaggc cgaggacgag 1500
gccgattact actgtagcag ctacaccagc tcctccacca gagtgtttgg caccggcacc 1560
aaagtgaccg tgctttaa 1578
<210> 151
<211> 2229
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
<400> 151
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatccagc 480
agcgcctcta caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgaccgtg 600
tcctggaact ctggcgctct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg acagtgccca gcagctctct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgcccccga actgctggga 840
ggcccttccg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 900
cccgaagtga cctgcgtggt ggtggatgtg tcccacgagg accctgaagt gaagtttaat 960
tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcctagaga ggaacagtac 1020
gccagcacct accgggtggt gtccgtgctg acagtgctgc accaggactg gctgaacggc 1080
aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 1140
agcaaggcca agggccagcc ccgcgaaccc caggtgtaca cactgccccc aagcagggac 1200
gagctgacca agaaccaggt gtccctgacc tgtctcgtga aaggcttcta ccccagcgat 1260
atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 1320
gtgctggaca gcgacggctc attcttcctg tacagcaagc tgaccgtgga caagtcccgg 1380
tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1440
acccagaagt ccctgagcct gagccccggc aagctcgagg aggtgcagct gctggaatct 1500
ggcggaggac ttgttcagcc tggcggctct ctgagactgt cttgtgccgc cagcggcttc 1560
accttcagca gctatatcat gatgtgggtc cgacaggccc ctggcaaagg ccttgaatgg 1620
gtgtccagca tctatcccag cggcggcatc accttttacg ccgacacagt gaagggcaga 1680
ttcaccatca gccgggacaa cagcaagaac accctgtacc tgcagatgaa cagcctgaga 1740
gccgaggaca ccgccgtgta ctactgcgcc agaatcaagc tgggcaccgt gaccaccgtg 1800
gattattggg gacagggcac cctggtcacc gtgtcctcca gatcttctac aaagggcccc 1860
aagctggaag agggcgagtt tagcgaagcc caattgcaga gcgccctgac acagcctgca 1920
tccgtgtctg gatctccagg ccagagcatc accatctctt gtaccggcac aagctccgat 1980
gtcggcggct acaattacgt gtcctggtat cagcagcacc ccggcaaggc ccctaagctg 2040
atgatctacg acgtgtccaa cagaccctcc ggcgtgtcca atagattcag cggcagcaag 2100
agcggcaaca ccgccagcct gacaattagc ggactgcagg ccgaggacga ggccgattac 2160
tactgtagca gctacaccag ctcctccacc agagtgtttg gcaccggcac caaagtgacc 2220
gtgctttaa 2229
<210> 152
<211> 1581
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK)-Fab2-scFv:PD-L1(Ave)
<400> 152
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatcctct 480
agcgccagca caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgacagtg 600
tcctggaact ctggcgccct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg actgtgccca gcagcagcct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgccctcga ggaggtgcag 840
ctgctggaat ctggcggagg acttgttcag cctggcggct ctctgagact gtcttgtgcc 900
gccagcggct tcaccttcag cagctatatc atgatgtggg tccgacaggc ccctggcaaa 960
ggccttgaat gggtgtccag catctatccc agcggcggca tcacctttta cgccgacaca 1020
gtgaagggca gattcaccat cagccgggac aacagcaaga acaccctgta cctgcagatg 1080
aacagcctga gagccgagga caccgccgtg tactactgcg ccagaatcaa gctgggcacc 1140
gtgaccaccg tggattattg gggacagggc accctggtca ccgtgtcctc cagatcttct 1200
acaaagggcc ccaagctgga agagggcgag tttagcgaag cccaattgca gagcgccctg 1260
acacagcctg catccgtgtc tggatctcca ggccagagca tcaccatctc ttgtaccggc 1320
acaagctccg atgtcggcgg ctacaattac gtgtcctggt atcagcagca ccccggcaag 1380
gcccctaagc tgatgatcta cgacgtgtcc aacagaccct ccggcgtgtc caatagattc 1440
agcggcagca agagcggcaa caccgccagc ctgacaatta gcggactgca ggccgaggac 1500
gaggccgatt actactgtag cagctacacc agctcctcca ccagagtgtt tggcaccggc 1560
accaaagtga ccgtgcttta a 1581
<210> 153
<211> 2208
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗PD-L1(Ave)-IgG2-VH-scFv:41BB(HBBK)
<400> 153
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attcgaggtg 120
cagctgctgg aatctggcgg aggacttgtt cagcctggcg gctctctgag actgtcttgt 180
gccgccagcg gcttcacctt cagcagctat atcatgatgt gggtccgaca ggcccctggc 240
aaaggccttg aatgggtgtc cagcatctat cccagcggcg gcatcacctt ttacgccgac 300
acagtgaagg gcagattcac catcagccgg gacaacagca agaacaccct gtacctgcag 360
atgaacagcc tgagagccga ggacaccgcc gtgtactact gcgccagaat caagctgggc 420
accgtgacca ccgtggatta ttggggacag ggcaccctgg tcaccgtgtc ctccagatcc 480
tcgagtgcta gcaccaaggg cccatcggtc ttccccctgg cgccctgctc caggagcacc 540
tccgagagca cagcggccct gggctgcctg gtcaaggact acttccccga accggtgacg 600
gtgtcgtgga actcaggcgc tctgaccagc ggcgtgcaca ccttcccagc tgtcctacag 660
tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcaa cttcggcacc 720
cagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagacagtt 780
gagcgcaaat gttgtgtcga gtgcccaccg tgcccagcac cacctgtggc aggaccgtca 840
gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 900
acgtgcgtgg tggtggacgt gagccacgaa gaccccgagg tccagttcaa ctggtacgtg 960
gacggcgtgg aggtgcataa tgccaagaca aagccacggg aggagcagtt caacagcacg 1020
ttccgtgtgg tcagcgtcct caccgttgtg caccaggact ggctgaacgg caaggagtac 1080
aagtgcaagg tctccaacaa aggcctccca gcccccatcg agaaaaccat ctccaaaacc 1140
aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 1200
aagaaccagg tcagcctgac ctgcctggtc aaaggcttct accccagcga catcgccgtg 1260
gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc catgctggac 1320
tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1380
gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1440
agcctctccc tgtctccggg taaagaattc caggtccagc tgcagcagtc tggcgccgaa 1500
gttattaagc ctggcgcctc cgtgaagctg agctgtaaag ccagcggcta caccttcagc 1560
agctactgga tgcactgggt ccgacaggct ccaggacaag gcctggaatg gatcggcgag 1620
atcaaccctg gcaacggcca caccaactac aacgagaagt tcaagagccg ggccacactg 1680
accggcgata ccagcacaag caccgtgtac atggaactga gcagcctgag aagcgaggac 1740
accgccgtgt actactgcgc cagatccttt accaccgcca gagcctttgc ctattggggc 1800
cagggaacac tggtcaccgt gtccagcaga tctagcacaa agggccccaa gctggaagag 1860
ggcgagttta gcgaggccca attggacatc gtgatgactc agagccccgc cttcctgtct 1920
gtgacccctg gcgagaaagt gaccatcacc tgtagagcca gccagaccat cagcgactac 1980
ctgcactggt atcagcagaa gcccgatcag gcccctaagc tgctgattaa gtacgcctct 2040
cagagcatca gcggcatccc cagcagattt tctggcagcg gctctggcac cgacttcacc 2100
tttaccatca gctccctgga agccgaggat gccgccacct actattgtca ggacggccac 2160
agcttccctc caacctttgg acagggcaca aagctggaaa tcaagtaa 2208
<210> 154
<211> 783
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗PD-L1(Ave)-VL
<400> 154
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagagc 120
gccctgacac agcctgcatc cgtgtctgga tctccaggcc agagcatcac catctcttgt 180
accggcacaa gctccgatgt cggcggctac aattacgtgt cctggtatca gcagcacccc 240
ggcaaggccc ctaagctgat gatctacgac gtgtccaaca gaccctccgg cgtgtccaat 300
agattcagcg gcagcaagag cggcaacacc gccagcctga caattagcgg actgcaggcc 360
gaggacgagg ccgattacta ctgtagcagc tacaccagct cctccaccag agtgtttggc 420
accggcacca aagtgaccgt gcttggatcc gaaatcaagc gtacggtggc cgctcccagc 480
gtgttcatct tcccacctag cgacgagcag ctgaagtccg gcacagcctc tgtcgtgtgc 540
ctgctgaaca acttctaccc ccgcgaggcc aaggtgcagt ggaaggtgga caatgccctg 600
cagagcggca acagccagga aagcgtgacc gagcaggaca gcaaggactc cacctacagc 660
ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgc 720
gaagtgaccc accagggcct gtctagcccc gtgaccaaga gcttcaaccg gggcgagtgc 780
taa 783
<210> 155
<211> 2223
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗muCD27-IgG1(N297A)-VH-scFv:Fn14(18D1)
<400> 155
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaacttgtg aagcctggca gcagcgtgaa gatcagctgt 180
aaagccagcg gctacacctt caccagctac gacatgcact ggatcaagca gcagcccggc 240
aaaggcctgg aatggatcgg ctggatctac cccggcaacg gcaacaccaa gtacaaccag 300
aagttcaacg gcaaggccac actgaccgcc gacatctcta gcagcacagc ctacatgcag 360
ctgagcagcc tgaccagcga agatagcgcc gtgtacttct gcgccaaatg gggctacaac 420
aacttcgact actggggcca gggcgtgatg gtcaccgtgt ctagcagatc cagcagcgcc 480
tctacaaagg gccccagcgt gttccctctg gcccctagca gcaagagcac atctggcgga 540
acagccgccc tgggctgcct cgtgaaggac tactttcccg agcccgtgac cgtgtcctgg 600
aactctggcg ctctgacaag cggcgtgcac acctttccag ccgtgctgca gagcagcggc 660
ctgtactctc tgagcagcgt cgtgacagtg cccagcagct ctctgggcac ccagacctac 720
atctgcaacg tgaaccacaa gcccagcaac accaaggtgg acaagaaggt ggaacccaag 780
agctgcgaca agacccacac ctgtccccct tgtcctgccc ccgaactgct gggaggccct 840
tccgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 900
gtgacctgcg tggtggtgga tgtgtcccac gaggaccctg aagtgaagtt taattggtac 960
gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacgccagc 1020
acctaccggg tggtgtccgt gctgacagtg ctgcaccagg actggctgaa cggcaaagag 1080
tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 1140
gccaagggcc agccccgcga accccaggtg tacacactgc ccccaagcag ggacgagctg 1200
accaagaacc aggtgtccct gacctgtctc gtgaaaggct tctaccccag cgatatcgcc 1260
gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1320
gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagtc ccggtggcag 1380
cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1440
aagtccctga gcctgagccc cggcaagctc gaggaggtgc agctggtgga atctggcggc 1500
ggactggtgc agcctggcgg atctctgaga ctgagctgtg ccgccagcgg cttcaccttc 1560
agcaactact ggatgagctg ggtgcgccag gcccctggca aaggactgga atgggtgtcc 1620
ggcatcaacc caggcggcac ctctacctac tacgccgaca gcgtgaaggg ccggttcacc 1680
atcagccggg acaacgccaa gaacaccctg tacctgcaga tgaacagcct gaagtccgag 1740
gacaccgccg tgtactactg cgccaagcac ctgggcaact ggggcgagta caattactgg 1800
ggccagggca cacaagtgac cgtgtccagt agatctagca ccaagggccc caagctggaa 1860
gagggcgagt tcagcgaggc ccaattgcag agcgccctga cccagcctcc aagcgtgtca 1920
ggctctcctg gcaagaccgt gaccatcagc tgtgctggca ccggcggaga tgtgggctac 1980
agaaacagcg tgtcctggta tcagcagctg cccggcatgg cccccaaact gctgatctac 2040
gacgtggaca agcgggcctc tggcatcacc gacagattca gcggcagcaa gagcggcgat 2100
accgccagcc tgacaatcag cggagtgcag agcgaggacg aggccgacta ctactgtgcc 2160
agccagagaa gcggaatcgc cgccgtgttt ggcggaggca cacacctgac agtgctgggc 2220
taa 2223
<210> 156
<211> 774
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗muCD27-VL
<400> 156
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
cagatgacac agagccctgc cagcctgtct gcctctctgg gagagacagt gtccatcgat 180
tgtctggcca gcgagggcat cagcaacgac ctggcttggt atcagcagaa gtccggcaag 240
agccctcagc tgctgatcaa cagcgccagc agactggaag atggcgtgcc cagcagattt 300
tctggctctg gcagcggcac ccggtacagc ctgaagattt ctggcatgca gcccgaggac 360
gaggccgaat acttctgcct gcaaagctac agaagcccct ggacctttgg cggaggcaca 420
aagctggaac tgaagggatc cgaaatcaag cgtacggtgg ccgctcccag cgtgttcatc 480
ttcccaccta gcgacgagca gctgaagtcc ggcacagcct ctgtcgtgtg cctgctgaac 540
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaatgccct gcagagcggc 600
aacagccagg aaagcgtgac cgagcaggac agcaaggact ccacctacag cctgagcagc 660
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaagtgacc 720
caccagggcc tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaa 774
<210> 157
<211> 750
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 146所示的蛋白质构建体
<400> 157
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Gly Ser Ile Ser Ser Gly Asp Tyr Phe Trp Ser Trp Ile Arg Gln Leu
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Ile Gly His Ile His Asn Ser Gly Thr
85 90 95
Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp
100 105 110
Thr Ser Lys Lys Gln Phe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala
115 120 125
Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Gly Gly Asp Tyr Tyr
130 135 140
Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
145 150 155 160
Arg Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
165 170 175
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
180 185 190
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
195 200 205
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
210 215 220
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
225 230 235 240
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
245 250 255
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
260 265 270
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
275 280 285
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
290 295 300
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
305 310 315 320
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
325 330 335
Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val
340 345 350
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
355 360 365
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
370 375 380
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
385 390 395 400
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
405 410 415
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
420 425 430
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
435 440 445
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
450 455 460
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
465 470 475 480
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu
485 490 495
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Met Gln Pro Gly Gly
500 505 510
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser
515 520 525
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
530 535 540
Ser Ser Ile Tyr Ser Asp Ser Ser Tyr Thr Tyr Tyr Ala Asp Ser Val
545 550 555 560
Lys Ser Arg Phe Thr Ile Ser Thr Asp Asn Ala Lys Asn Thr Leu Tyr
565 570 575
Leu Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Val Tyr Tyr Cys
580 585 590
Ala Gly Ser Ser Asp Tyr Glu Gly Ser Phe Ala Ser Trp Gly Gln Gly
595 600 605
Thr Gln Val Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu
610 615 620
Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu Gln Ser Val Val Thr Gln
625 630 635 640
Pro Pro Ser Leu Ser Ala Ser Pro Gly Ser Ser Val Arg Leu Thr Cys
645 650 655
Thr Leu Ser Ser Gly Asn Ser Val Gly Asn Tyr Asp Ile Ser Trp Tyr
660 665 670
Gln Gln Lys Ala Gly Ser Pro Pro Arg Tyr Leu Leu Tyr Tyr Tyr Ser
675 680 685
Asp Ser Val Lys His Gln Gly Ser Gly Val Pro Ser Arg Phe Ser Gly
690 695 700
Ser Ser Asp Ala Ser Ala Asn Ala Gly Leu Leu Leu Ile Ser Gly Leu
705 710 715 720
Gln Pro Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ala Tyr Lys Ser Gly
725 730 735
Ser His Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly
740 745 750
<210> 158
<211> 255
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 147所示的蛋白质构建体
<400> 158
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
35 40 45
Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser
50 55 60
Gln Gly Ile Ser Arg Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly
65 70 75 80
Gln Ala Pro Ser Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly
85 90 95
Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
100 105 110
Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln
115 120 125
Gln Phe Gly Ser Ser Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Gly
130 135 140
Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
145 150 155 160
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
165 170 175
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
180 185 190
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
195 200 205
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
210 215 220
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
225 230 235 240
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
245 250 255
<210> 159
<211> 741
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 148所示的蛋白质构建体
<400> 159
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Val Gln Ser Gly Ala Glu
35 40 45
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Thr Ala Ser Gly
50 55 60
Phe Asn Ile Lys Asp Tyr Tyr Val His Trp Val Lys Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Met Gly Arg Ile Asp Pro Glu Asp Gly Asp Ser
85 90 95
Lys Tyr Ala Pro Lys Phe Gln Gly Lys Ala Thr Met Thr Ala Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Thr Thr Ser Tyr Tyr Val Gly Thr Tyr Gly
130 135 140
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Ser
145 150 155 160
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
165 170 175
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
180 185 190
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
195 200 205
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
210 215 220
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
225 230 235 240
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
245 250 255
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
260 265 270
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
275 280 285
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
290 295 300
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
305 310 315 320
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
325 330 335
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
340 345 350
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
355 360 365
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
370 375 380
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
385 390 395 400
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
405 410 415
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
420 425 430
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
435 440 445
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
450 455 460
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
465 470 475 480
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln Leu
485 490 495
Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu
500 505 510
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ile Met Met Trp
515 520 525
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Tyr
530 535 540
Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val Lys Gly Arg Phe
545 550 555 560
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn
565 570 575
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ile Lys
580 585 590
Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu Val
595 600 605
Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly
610 615 620
Glu Phe Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Ala Ser
625 630 635 640
Val Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr
645 650 655
Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His
660 665 670
Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg Pro
675 680 685
Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala
690 695 700
Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr
705 710 715 720
Cys Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly Thr
725 730 735
Lys Val Thr Val Leu
740
<210> 160
<211> 256
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 149所示的蛋白质构建体
<400> 160
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
35 40 45
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Ser Ala Ser
50 55 60
Ser Ser Val Ser Tyr Met Leu Trp Phe Gln Gln Lys Pro Gly Lys Ala
65 70 75 80
Pro Lys Leu Leu Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro
85 90 95
Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
100 105 110
Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg
115 120 125
Thr Phe Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
130 135 140
Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
145 150 155 160
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
165 170 175
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
180 185 190
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
195 200 205
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
210 215 220
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
225 230 235 240
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
245 250 255
<210> 161
<211> 525
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 150所示的蛋白质构建体
<400> 161
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Val Gln Ser Gly Ala Glu
35 40 45
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Thr Ala Ser Gly
50 55 60
Phe Asn Ile Lys Asp Tyr Tyr Val His Trp Val Lys Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Met Gly Arg Ile Asp Pro Glu Asp Gly Asp Ser
85 90 95
Lys Tyr Ala Pro Lys Phe Gln Gly Lys Ala Thr Met Thr Ala Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Thr Thr Ser Tyr Tyr Val Gly Thr Tyr Gly
130 135 140
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Ser
145 150 155 160
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
165 170 175
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
180 185 190
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
195 200 205
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
210 215 220
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
225 230 235 240
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
245 250 255
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
260 265 270
Pro Ala Leu Glu Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val
275 280 285
Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
290 295 300
Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly
305 310 315 320
Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr
325 330 335
Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
340 345 350
Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
355 360 365
Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp
370 375 380
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Thr
385 390 395 400
Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu Gln
405 410 415
Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser
420 425 430
Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn
435 440 445
Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met
450 455 460
Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser
465 470 475 480
Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln
485 490 495
Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser Ser
500 505 510
Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu
515 520 525
<210> 162
<211> 742
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 151所示的蛋白质构建体
<400> 162
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
275 280 285
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
290 295 300
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
305 310 315 320
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
325 330 335
Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
340 345 350
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
355 360 365
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
370 375 380
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
385 390 395 400
Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
405 410 415
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
420 425 430
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
435 440 445
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
450 455 460
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
465 470 475 480
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln
485 490 495
Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
500 505 510
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ile Met Met
515 520 525
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile
530 535 540
Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val Lys Gly Arg
545 550 555 560
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
565 570 575
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ile
580 585 590
Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu
595 600 605
Val Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu
610 615 620
Gly Glu Phe Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Ala
625 630 635 640
Ser Val Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly
645 650 655
Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln
660 665 670
His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg
675 680 685
Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr
690 695 700
Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr
705 710 715 720
Tyr Cys Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly
725 730 735
Thr Lys Val Thr Val Leu
740
<210> 163
<211> 526
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 152所示的蛋白质构建体
<400> 163
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Leu Glu Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu
275 280 285
Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
290 295 300
Thr Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys
305 310 315 320
Gly Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe
325 330 335
Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser
340 345 350
Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
355 360 365
Ala Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val
370 375 380
Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
385 390 395 400
Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu
405 410 415
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
420 425 430
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr
435 440 445
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
450 455 460
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe
465 470 475 480
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
485 490 495
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
500 505 510
Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu
515 520 525
<210> 164
<211> 735
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 153所示的蛋白质构建体
<400> 164
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Glu Val Gln Leu Leu Glu Ser Gly Gly Gly
35 40 45
Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
50 55 60
Phe Thr Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Lys Gly Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr
85 90 95
Phe Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
100 105 110
Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr
130 135 140
Val Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser
145 150 155 160
Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys
165 170 175
Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys
180 185 190
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
195 200 205
Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
210 215 220
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr
225 230 235 240
Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val
245 250 255
Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro
260 265 270
Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
275 280 285
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
290 295 300
Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val
305 310 315 320
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
325 330 335
Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln
340 345 350
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly
355 360 365
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
370 375 380
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
385 390 395 400
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
405 410 415
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
420 425 430
Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
435 440 445
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
450 455 460
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
465 470 475 480
Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln
485 490 495
Ser Gly Ala Glu Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys
500 505 510
Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg
515 520 525
Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly
530 535 540
Asn Gly His Thr Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu
545 550 555 560
Thr Gly Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu
565 570 575
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr
580 585 590
Ala Arg Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
595 600 605
Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser
610 615 620
Glu Ala Gln Leu Asp Ile Val Met Thr Gln Ser Pro Ala Phe Leu Ser
625 630 635 640
Val Thr Pro Gly Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Thr
645 650 655
Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro
660 665 670
Lys Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser
675 680 685
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser
690 695 700
Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asp Gly His
705 710 715 720
Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
725 730 735
<210> 165
<211> 260
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 154所示的蛋白质构建体
<400> 165
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Ser Ala Leu Thr Gln Pro Ala Ser Val
35 40 45
Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser
50 55 60
Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro
65 70 75 80
Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg Pro Ser
85 90 95
Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser
100 105 110
Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys
115 120 125
Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly Thr Lys
130 135 140
Val Thr Val Leu Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
145 150 155 160
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
165 170 175
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
180 185 190
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
195 200 205
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
210 215 220
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
225 230 235 240
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
245 250 255
Arg Gly Glu Cys
260
<210> 166
<211> 740
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 155所示的蛋白质构建体
<400> 166
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Leu Val Lys Pro Gly Ser Ser Val Lys Ile Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Thr Ser Tyr Asp Met His Trp Ile Lys Gln Gln Pro Gly
65 70 75 80
Lys Gly Leu Glu Trp Ile Gly Trp Ile Tyr Pro Gly Asn Gly Asn Thr
85 90 95
Lys Tyr Asn Gln Lys Phe Asn Gly Lys Ala Thr Leu Thr Ala Asp Ile
100 105 110
Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp
115 120 125
Ser Ala Val Tyr Phe Cys Ala Lys Trp Gly Tyr Asn Asn Phe Asp Tyr
130 135 140
Trp Gly Gln Gly Val Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala
145 150 155 160
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
165 170 175
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
180 185 190
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
195 200 205
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
210 215 220
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
225 230 235 240
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
245 250 255
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
260 265 270
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
275 280 285
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
290 295 300
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
305 310 315 320
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
325 330 335
Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
340 345 350
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
355 360 365
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
370 375 380
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
385 390 395 400
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
405 410 415
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
420 425 430
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
435 440 445
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
450 455 460
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
465 470 475 480
Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln Leu Val
485 490 495
Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser
500 505 510
Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr Trp Met Ser Trp Val
515 520 525
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile Asn Pro
530 535 540
Gly Gly Thr Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr
545 550 555 560
Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser
565 570 575
Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys His Leu Gly
580 585 590
Asn Trp Gly Glu Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
595 600 605
Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe
610 615 620
Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Pro Ser Val Ser
625 630 635 640
Gly Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Ala Gly Thr Gly Gly
645 650 655
Asp Val Gly Tyr Arg Asn Ser Val Ser Trp Tyr Gln Gln Leu Pro Gly
660 665 670
Met Ala Pro Lys Leu Leu Ile Tyr Asp Val Asp Lys Arg Ala Ser Gly
675 680 685
Ile Thr Asp Arg Phe Ser Gly Ser Lys Ser Gly Asp Thr Ala Ser Leu
690 695 700
Thr Ile Ser Gly Val Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala
705 710 715 720
Ser Gln Arg Ser Gly Ile Ala Ala Val Phe Gly Gly Gly Thr His Leu
725 730 735
Thr Val Leu Gly
740
<210> 167
<211> 257
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 156所示的蛋白质构建体
<400> 167
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Met Thr Gln Ser Pro Ala Ser
35 40 45
Leu Ser Ala Ser Leu Gly Glu Thr Val Ser Ile Asp Cys Leu Ala Ser
50 55 60
Glu Gly Ile Ser Asn Asp Leu Ala Trp Tyr Gln Gln Lys Ser Gly Lys
65 70 75 80
Ser Pro Gln Leu Leu Ile Asn Ser Ala Ser Arg Leu Glu Asp Gly Val
85 90 95
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Arg Tyr Ser Leu Lys
100 105 110
Ile Ser Gly Met Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Leu Gln
115 120 125
Ser Tyr Arg Ser Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu
130 135 140
Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
145 150 155 160
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
165 170 175
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
180 185 190
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
195 200 205
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
210 215 220
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
225 230 235 240
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
245 250 255
Cys
具体实施方式
图1：TNFSF和TNFRSF的配体和受体。
图2：受体特异性IgG引起的TNFRSF受体的FcγR依赖的刺激。用鼠FcγR2B的表达质粒或空载体转染的Hek293细胞与所示细胞系的细胞共培养，所示细胞系中所示TNFRSF受体的刺激导致IL8的产生。在与各种TNFRSF受体特异性IgG(指示克隆名称)孵育过夜后，使用ELISA捕获IL8的产生。请注意，TNFR1和LTβR是已经被可溶性配体三聚体强烈激活的TNFRSF成员示例，并且在不表达FcγR的细胞中，它们也可以被受体特异性抗体强烈激活。所示的其他TNFRSF受体结合可溶性TNF配体没有或仅有限激活，并且在不存在FcγR结合的情况下不被抗体激活或仅低效地激活。然而，这些抗TNFRSF受体在抗体与FcγR结合后被强烈激活。
材料与方法：用编码鼠FcγR2B或空载体的表达质粒pCMV-SPORT6(SourceBicoscience)瞬时转染不表达(或仅适度表达)所示TNFRSF受体和FcγR的Hek293细胞(ATCC)。如Kums et al.,MAbs.2017Apr；9(3):506-520中所述，使用聚乙烯亚胺(PEI)将FcγR表达质粒或空载体转染Hek293细胞。第二天，收获Hek293转染子，将20.000个细胞的等分试样加入到前一天已接种(10000–20000个细胞/孔)的96孔板的孔中，这些细胞可以响应目标TNFRSF受体的激活并产生IL8。然后将共培养物用所示的抗TNFRSF受体抗体(3μg/ml)刺激过夜，一式三份，最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图3：受体特异性IgG引起的TNFRSF受体TNFR2的FcγR依赖的刺激。用所示FcγR2B的表达质粒或空载体转染的Hek293细胞与刺激TNFR2诱导产生IL8的细胞共培养。在与所示TNFR2特异性IgG孵育过夜后，使用ELISA捕获IL8的产生。
材料与方法：用编码人活性FcγR CD32A(FcγR2IIA)、人抑制性FcγR CD32B(FcγRIIB)或空载体的表达质粒pCMV-SPORT6(来自Bicoscience)瞬时转染不表达TNFR2或FcγR的Hek293细胞(ATCC)。如Kums et al.,MAbs.2017Apr；9(3):506-520中所述，使用聚乙烯亚胺(PEI)将FcγR表达质粒或空载体转染Hek293细胞。第二天，收获Hek293转染子，将20.000个细胞的等分试样加入到前一天已接种稳定表达TNFR2的HT1080细胞(密度为20000个细胞/孔)的96孔板的孔中。然后将共培养物用所示的抗人TNFR2抗体(1μg/ml)刺激过夜，一式三份，所述抗体由自身生成并生产。显示了被抗体识别的TNFR2的富含半胱氨酸的结构域(CRD)亚结构域(CRD1-CRD4)。所使用的细胞中TNFR2的激活导致增强的IL8产生。因此，为了确定TNFR2活性，最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图4：抗CD40 IgG1 G28.5的scBaff融合蛋白与BaffR、TACI或BCMA结合后，显示出增强的CD40刺激活性。(A)融合蛋白的结构。(B)空载体与分别编码BaffR、BCMA和TACI的表达质粒转染Hek293细胞。第二天，将转染子与HT1080-CD40细胞共培养，所述细胞不表达BaffR、BCMA和TACI，但在CD40刺激后强烈产生IL8。用指示浓度的抗CD40(G28.5)-HC:scBaff刺激共培养物，一式三份。次日，在不同共培养物的上清液中测定人IL8的量。(C)将表达BaffR和TACI的L363细胞、Hek293细胞(BaffR、BCMA和TACI阴性)和HT1080-CD40细胞共培养，然后用指示浓度的抗CD40(G28.5)-HC:scBaff刺激共培养物。第二天，在共培养物的上清液中再次测定人IL8的量。请注意，scBaff中Baff的三个原聚体仅包含全长Baff分子的胞外TNF同源结构域。
材料与方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的Hek293细胞和FcgR表达载体，用编码BaffR、TACI和BCMA的表达质粒或空载体(＝阴性对照)瞬时转染不表达CD40、BaffR、TACI或BCMA的Hek293细胞。收获转染子，并将30.000个细胞的等分试样加入到前一天已以20000个细胞/孔的密度接种了HT1080-CD40细胞(稳定转染CD40的HT1080细胞)的96孔板的孔中。可选地，将HT1080-CD40细胞与50.000个细胞/孔的内源性表达BaffR和TACI的骨髓瘤细胞系L363共培养。所述HT1080-CD40细胞对CD40的激活产生大量响应。将共培养物用TRAAFFIAA抗CD40(G28.5)-IgG1(N297)-scBaff刺激过夜，一式三份，最后通过ELISA测定IL8的产量。
图5：抗CD95 IgG1 E09的scBaff融合蛋白与BaffR、TACI或BCMA结合后显示出增加的CD95刺激活性。(A)融合蛋白的结构。(B)空载体或编码BaffR、BCMA和TACI的表达质粒转染Hek293细胞。第二天，将转染子与HT1080细胞共培养，所述细胞不表达BaffR、BCMA和TACI，并在CD95刺激后发生凋亡。用指示浓度的抗CD95(E09)-HC:scBaff处理共培养物并在次日通过结晶紫染色评估粘附生长的HT1080细胞的活性。(C)将表达BaffR和TACI的L363细胞、Hek293细胞(BaffR、BCMA和TACI阴性)与HT1080细胞共培养。再次用抗CD95(E09)-HC:scBaff刺激共培养物过夜，最后通过结晶紫染色评估HT1080的活性。请注意，scBaff中Baff的三个原聚体仅包含全长Baff分子的胞外TNF同源结构域。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的Hek293细胞和FcgR表达载体，用编码BaffR、TACI和BCMA的表达质粒或空载体(＝阴性对照)瞬时转染不表达BaffR、TACI或BCMA和表达可忽略量的CD95的Hek293细胞。收获转染子，并将30.000个细胞的等分试样加入前一天已以20000细胞/孔的密度接种HT1080细胞的96孔板的孔中。可选地，将HT1080细胞与50.000细胞/孔的内源性表达BaffR和TACI的骨髓瘤细胞系L363共培养。在没有敏化剂的情况下，HT1080细胞对CD95诱导的细胞死亡敏感。将共培养物用TRAAFFIAA抗CD95(E09)-IgG1(N297)-scBaff刺激过夜，一式三份，最后通过粘附生长的HT1080细胞的结晶紫染色测定细胞死亡诱导。
图6：结合TNFR2的抗CD40-IgG1融合蛋白在结合TNFR2后具有增强的CD40刺激活性。将既不表达TNFR2也不表达CD40的HeLa细胞、在TNFR2刺激后仅产生中等水平IL8的HeLa-TNFR2转染子与既不表达TNFR2也不表达CD40的HT1080细胞以及在CD40刺激后产生非常高水平的IL8的HT1080-CD40转染子共培养，并用指示浓度的靶向TNFR2的抗CD40融合蛋白(抗CD40(G28.5)-HC:scTNF80)刺激。第二天，测定不同共培养物的上清液中的IL8的量。请注意，scTNF80中TNF80的三个原聚体仅包含全长鼠TNF分子的胞外TNF同源结构域，包含赋予TNFR2特异性的突变。如前所述，以96孔形式进行测定，一式三份。
图7：结合CD19的抗CD95-IgG1融合蛋白与表达CD19的细胞共培养时，在HT1080细胞中显示增强的细胞死亡诱导。将不表达CD19的CD95敏感细胞系HT1080与Jurkat(CD19阴性)细胞和BJAB(CD19阳性)细胞共培养，并与结合CD19的抗CD95融合蛋白(抗CD95(E09)-HC:scFv:CD19)以指示浓度孵育。第二天，通过结晶紫染色测定粘附生长的HT1080细胞的活性。
材料与方法：实验在96孔板上进行，一式三份。HT1080细胞按每孔20000个细胞接种。接下来加入Jurkat和BJAB细胞(每孔30000个)以及指示浓度的抗CD95(E09)-HC:scFv:CD19。通过结晶紫染色贴壁生长的HT1080细胞测定细胞死亡诱导。借助于未处理的HT1080细胞(＝100％活性)和用含有CHX、Velcade和Fc-CD95L的高毒性混合物处理的HT1080细胞(＝0活性)对细胞活性归一化。
图8：TNFR2特异性抗体C4的IL2融合蛋白以依赖IL2R的方式增强刺激TNFR2。将既不表达TNFR2也不表达IL2R的Hek293细胞用编码IL2R的亚基的表达质粒的混合物或者空载体转染。然后将Hek293转染子与表达TNFR2的HeLa转染子共培养，并用指示浓度的抗TNFR2(C4)-HC:IL2刺激。第二天，使用ELISA捕获可通过TNFR2诱导产生的IL8。
材料与方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码IL-2受体亚基的表达质粒混合物瞬时转染Hek293细胞，或空载体作为阴性对照瞬时转染Hek293细胞。收获转染子，将20.000个细胞的等分试样加入前一天已以20000个细胞/孔的密度接种了HeLa-TNFR2细胞的96孔板的孔中。所述HeLa-TNFR2细胞响应TNFR2的激活而产生IL8。将共培养物用TRAAFFIAA抗TNFR2(C4)-HC:IL2刺激过夜，一式三份，最后通过ELISA测定IL8的产生。
图9：TNFR2特异性抗体C4的锚定GITR和41BB的融合蛋白在结合GITR和41BB后显示出增强的TNFR2活化。(A)所使用的刺激TNFR2的锚定人GITR和鼠4-1BB的TRAAFFIAA的域结构。(B，C)用空载体或编码人GITR(B)或鼠41BB(C)的表达质粒转染既不表达GITR也不表达41BB的细胞。然后将转染子与响应TNFR2激活而产生IL8的HeLa-TNFR2细胞共培养。将共培养物用指示浓度的抗TNFR2(C4)-HC:scGITRL(B)和抗TNFR2(C4)-HC:sc(mu)4-1BBL(C)刺激。第二天，采用IL8特异性ELISA测定IL8的产生。(D)所使用的鼠GITR锚定的TNFR2刺激的TRAAFFIAA的域结构。(E)将用空载体或鼠GITR的表达质粒转染的细胞与HeLa-TNFR2细胞共培养。在用抗TNFR2(C4)-HC:(mu)GITRL刺激后，通过ELISA监测第二天IL8的产生。请注意，与人GITRL以及TNFSF的所有其他人和鼠配体相反，鼠GITRL是二聚体分子，并与两个而不是三个受体分子相互作用。因此，将鼠GITRL的单个原聚体融合到重链的C末端，以构建单个(mu)GITRL二聚体，其作为相应抗体-(mu)GITRL的锚定结构域。相反，使用scTNFSF配体(例如scGITRL、sc(mu)41BBL和scTNF80)作为锚定结构域融合至重链，使得每个抗体融合蛋白产生两个锚定结构域。请注意，scGITRL和sc(mu)41BBL中的GITRL和(mu)41BBL的三个原聚体仅包含全长配体分子的胞外TNF同源结构域。同样，(mu)GITRL仅指鼠全长鼠GITRL的THD。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码人或鼠GITR或鼠41BB的表达质粒或空载体(＝阴性对照)瞬时转染不表达GITR、41BB或TNFR2的Hek293细胞。收获转染子，将20000个细胞的等分试样加入已以20000个细胞/孔的密度接种HeLa-TNFR2细胞的96孔板的孔中。所述HeLa-TNFR2细胞响应TNFR2激活而产生IL8。将共培养物用所示的TRAAFFIAA刺激过夜，一式三份，最后通过ELISA测定IL8的产生。
图10：锚定细胞表面抗原CD20的CD40特异性抗体G28.5的scFv融合蛋白以依赖CD20的方式增强刺激CD40。将CD20阴性Jurkat细胞和CD20阳性BJAB细胞与可应答CD40的HT1080-CD40细胞共培养，并用指示浓度的抗CD40(G28.5)-HC:scFv:CD20刺激。第二天，使用ELISA捕获可通过CD40诱导产生的IL8。使用具有N297A突变的IgG1分子的变体，所述变体在体内具有抑制Fc结构域的ADCC活性。
材料与方法：实验在96孔板上进行，一式三份。以每孔20000个细胞接种HT1080-CD40细胞。第二天加入Jurkat(CD20-)和BJAB细胞(CD20+)(每孔30000个)，以及指定浓度的抗CD40-HC:scFv:CD20。最后通过ELISA(BD Biosciences)评估共培养上清液的IL8含量。
图11：抗体E09的抗CD95-IgG1 Fab2片段的scFv:CD20融合蛋白在与表达CD20的细胞共培养时，在HT1080细胞中显示出增强的细胞死亡诱导。将CD95敏感细胞系HT1080与Jurkat(CD20阴性)细胞和BJAB细胞(CD20阳性)共培养。用指示浓度的结合CD20的抗CD95融合蛋白抗CD95-Fab2-HC:scFv:CD20处理共培养物。第二天，通过结晶紫染色确定粘附生长的HT1080细胞的生存力。
材料与方法：实验在96孔板上进行，一式三份。以每孔接种20000个细胞接种HT1080细胞。接下来共同添加Jurkat和BJAB细胞(每孔30000个)以及指示浓度的TRAAFFIAA。借助于未处理的HT1080细胞(＝100％活性)和用含有CHX、Velcade和Fc-CD95L(＝0活性)的高毒性混合物处理的HT1080细胞对HT1080的细胞活性归一化。
图12：TNFR2特异性抗体C4的靶向细胞表面抗原的scFv融合蛋白以依赖抗原的方式增强刺激TNFR2。将CD19和CD20阴性的Jurkat细胞以及CD19和CD20阳性的Raji细胞与表达TNFR2的HeLa转染子共培养，并分别用指示浓度的抗TNFR2-IgG1(N297A)-HC:scFv:CD19(上图)或抗TNFR2-IgG1(N297A)-HC:scFv:CD20(下图)刺激。第二天，使用ELISA捕获可通过TNFR2诱导的IL8的产生。使用了不能触发ADCC的IgG1变体N297A。这里使用的亲本TNFR2特异性mAb C4是内部制备的。
材料与方法：实验在96孔板上进行，一式三份。上图：以每孔20000个细胞接种HeLa-TNFR2细胞。第二天，加入Jurkat(CD19-/CD20-)和RAJI细胞(CD19-/CD20-)(每孔30000)以及指示浓度的抗TNFR2-IgG1(N297A)-HC:scFv:CD19(上图)或者抗TNFR2-IgG1(N297A)-HC:scFv:CD20(下图)。最后通过ELISA(BD Biosciences)分析评估共培养上清液中由TNFR2激活产生的IL8含量。
图13：抗CD40 G28.5的scBaff融合蛋白的生化性质。(A)纯化的抗CD40(G28.5)-IgG1(N297)(I)、抗CD40(G28.5)-IgG1(N297)-HC:scBaff(II)、抗CD40(G28.5)-FAB2(III)和抗CD40(G28.5)-FAB2-HC:scBaff(IV)通过SDS-PAGE分离。然后通过可识别各种抗体融合蛋白的重链和轻链中存在的Flag表位的mAb M2进行Western blotting(左图)或银染(右图)对蛋白进行可视化。请注意，构建体II的两条链的大小非常相似，因此无法通过SDS-PAGE分离。(B)标记蛋白混合物和蛋白I至IV的凝胶过滤分析。(C)蛋白质I至IV的示意图。
材料与方法：抗体融合蛋白的制备和纯化如Kums et al.,SDS-PAGE,2017所述，银染和western blotting的方法如Lang et al.,2016所述。
Kums J,Nelke J,RüthB,
Lang I,Füllsack S,Wyzgol A,Fick A,Trebing J,Arana JA,Weisenberger D,Wajant H,Binding Studies of TNF Receptor Superfamily(TNFRSF)Receptors on Intact Cells.J Bio Chem.2016Mar 4；291(10):5022-37。
图14：抗CD40 G28.5的IgG1(N297A)与FAB2变体的scBaff融合蛋白在与内源性表达BaffR和TACI的淋巴瘤/骨髓瘤细胞结合后，表现出增强的CD40刺激活性。将MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞或缺少所有Baff结合受体表达的Jurkat细胞作为对照，与不表达BaffR、BCMA和TACI但在CD40刺激后会强烈产生IL8的HT1080-CD40细胞共培养。用指示浓度的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff或抗CD40(G28.5)-FAB2-HC:scBaff刺激共培养物，一式三份。第二天，在不同共培养物的上清液中测定人IL8的量。
材料与方法：HT1080-CD40细胞在96孔板中培养过夜(每孔20.000个细胞)。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat细胞，并以指示浓度的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff刺激所得共培养物过夜，一式三份。然后通过ELISA分析细胞上清液中产生的IL8。请注意，与HT1080-CD40细胞相比，MM1S、L363、BJAB和Jurkat细胞不产生IL8或产生可忽略不计的IL8。
图15：与可溶性Baff(TNC-Baff)的竞争抑制了抗CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff中IL8的产生。HT1080-CD40细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞共培养，Jurkat细胞作为对照。请注意，HT1080-CD40和Jurkat细胞不表达Baff相互作用受体(BaffR、TACI、BCMA)。在存在和不存在过量可溶性TNC-Baff(5μg/ml)的情况下，用200ng/ml的抗CD40(G28.5)-IgG1(N297A)-HC:scBaff或抗CD40(G28.5)-FAB2-HC:scBaff刺激共培养，一式三份。第二天，在各种共培养物的上清液中测定人IL8的量。
材料与方法：HT1080-CD40细胞在96孔板中培养过夜(每孔20.000个细胞)。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat细胞，并在存在和不存在可溶性TNC-Baff(5μg/ml)的情况下，用200ng/ml CD40(G28.5)-IgG1(N297A)-HC:scBaff和抗CD40(G28.5)-FAB2-HC:scBaff刺激所得共培养物过夜，一式三份，所述可溶性TNC-Baff是含有Tenascin-C三聚结构域的可溶性Baff的稳定形态(Berg et al.,2007)。然后通过ELISA分析细胞上清液中IL8的产生。请注意，与HT10890-CD40细胞相比，MM1S、L363、BJAB和Jurkat细胞不产生IL8或产生可忽略不计的IL8。
Berg D,Lehne M,MüllerN,Siegmund D,Münkel S,Sebald W,Pfizenmaier K,Wajant H,Enforced covalent trimerization increases the activity of the TNFligand family members TRAIL and CD95L,Cell Death Differ.2007Dec；14(12):2021-34,EPUB 2007Aug 17。
图16：抗CD95 IgG1 E09或其FAB2片段的scBaff融合蛋白与具有BaffR和TACI内源性表达的淋巴瘤/骨髓瘤悬浮细胞结合后，诱导贴壁HT1080细胞死亡。将贴壁生长的HT1080细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)悬浮细胞共培养，Jurkat细胞作为对照。请注意，HT1080和Jurkat细胞不表达与Baff相互作用的受体(BaffR、TACI、BCMA)。在1μg/ml CHX的存在下，用指示浓度的抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激共培养物，一式三份。第二天，通过剩余附着的HT1080细胞的结晶紫染色确定细胞活力。
材料与方法：HT1080细胞的接种密度为20.000个细胞/孔。第二天，每孔添加20.000个MM1S、L363、BJAB或Jurkat悬浮细胞，并将所得的共培养物用1μg/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激过夜，一式三份。最后，通过贴壁生长的HT1080细胞的结晶紫染色确定细胞死亡诱导。借助未经处理的HT1080细胞(＝100％活力)和以0.03％叠氮、50μg/ml CHX和500ng/ml Fc-CD95L(＝0％活力)的高毒性混合物处理的HT1080细胞的活力进行了归一化。
图17：与可溶性Baff(TNC-Baff)的竞争抑制了HT1080细胞和Baff相互作用受体表达细胞的共培养物中抗CD95(E09)-IgG1(N297A)-HC:scBaff和抗CD95(E09)-FAB2-HC:scBaff诱导的HT1080杀伤。HT1080细胞与MM1S(表达BCMA和中度TACI)、L363(表达BCMA和中度TACI)和BJAB(表达BaffR)细胞共培养，Jurkat细胞作为对照。请注意，HT1080-CD40和Jurkat细胞不表达Baff相互作用受体(BaffR、TACI、BCMA)。在存在和不存在过量的可溶性TNC-Baff(5μg/ml)的情况下，以1μg/ml CHX刺激共培养物与20ng/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff，一式三份。第二天，通过剩余附着的HT1080细胞的结晶紫染色确定细胞活力。
材料与方法：HT1080细胞的接种密度为20.000个细胞/孔。第二天，每孔加入w0.000个MM1S、L363、BJAB或Jurkat悬浮细胞，并在存在和不存在可溶性TNC-Baff(5μg/ml)的情况下，用20ng/ml抗CD95(E09)-IgG1(N297A)-HC:scBaff或抗CD95(E09)-FAB2-HC:scBaff刺激过夜共培养物，一式三份。最后，通过贴壁生长的HT1080细胞的结晶紫染色确定细胞死亡诱导。借助未经处理的HT1080细胞(＝00％活力)和以0.03％叠氮、50μg/ml CHX和500ng/ml Fc-CD95L(＝0％活力)的高毒性混合物处理的HT1080细胞的活力进行了归一化。TNC-Baff是包含tenascin-C的三聚结构域的可溶性Baff的一种稳定形态(Berg et al.,2007)。
Berg D,Lehne M,Müller N,Siegmund D,Münkel S,Sebald W,Pfizenmaier K,Wajant H.,Enforced covalent trimerization increases the activity of the TNFligand family members TRAIL and CD95L,Cell Death Differ,2007Dec；14(12):2021-34。
图18：抗CD95(E09)-IgG1(N297A)-HC:scBaff而不是抗CD95(E09)-IgG1(N297A)激活BaffR表达细胞中的凋亡半胱氨酸蛋白酶。将BJAB细胞(表达BaffR)和作为阴性对照的不表达任何Baff受体(BaffR、TACI、BCMA)的Jurkat细胞用指示浓度的抗CD95(E09)-IgG1(N297A)-HC:scBaff和抗CD95(E09)-IgG1(N297A)处理过夜。第二天，通过westernblotting分析总细胞裂解物的caspase-8的p18片段和caspase-3的p17片段的生成，两种片段是这些半胱氨酸蛋白酶的凋亡活化的指示剂。作为阳性对照，用500ng/ml Fc-CD95L处理Jurkat和BJAB细胞，所述两种细胞系均可被其诱导凋亡。
图19：IgG2同种型的TNFR2特异性抗体C4锚定细胞表面抗原CD20的scFv融合蛋白以依赖CD20的方式增强刺激TNFR2。将CD20阴性Jurkat和CD20阳性BJAB细胞与TNFR2反应性HeLa-TNFR2细胞共培养，并用指示浓度的抗TNFR2(C4)-IgG2-HC:scFv:CD20刺激。第二天，使用ELISA测定了可以经由TNFR2诱导产生的IL8。
材料与方法：以每孔20.000个HeLa-TNFR2转染子(Weiss et al.,1997)接种在96孔板中。第二天，加入CD20阳性BJAB细胞或作为阴性对照的20.000个缺乏内源性表达CD20的Jurkat细胞。然后用指示浓度的抗TNFR2(C4)-IgG2-HC:scFvCD20处理共培养物，次日通过ELISA测定IL8的产生。
Weiss T,Grell M,Hessabi B,Bourteele S,Müller G,Scheurich P,WajantH.v,Enhancement of TNF receptor p6-mediated cytotoxicity by TNF receptor p80:requirement of the TNF receptor-associated factor-2binding site,J Immunol,1997Mar 1；158(5):2398-404。
图20：IgG1同种型的TNFR2特异性抗体C4锚定细胞表面抗原CD70的scFv融合蛋白以依赖CD27L/CD70r方式增强刺激TNFR2。(A)融合蛋白的结构。(B，C)用空载体或编码CD27L(或称CD70)的表达质粒转染Hek293细胞。第二天，将转染子与在TNFR2表达时产生IL8的HeLa-TNFR2转染子共培养。用指示浓度的抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(1F6)(B)或抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(2H5)处理共培养物(C)。最后，通过ELISA测定IL8的产生。
材料和方法：如Kums et al.,MAbs.2017Apr；9(3):506-520中所述的，对于Hek293细胞和FcgR表达载体，用编码CD27L的表达质粒(＝阴性对照)瞬时转染不表达TNFR2或CD27L(CD70)的Hek293细胞。收获转染子，将20.000个细胞等分添加到前一天已以20.000个细胞/孔的密度接种了HeLa-TNFR2细胞的96孔板的孔中。用TRAAFFIAA，即抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(1F6)(B)和抗TNFR2(C4)-IgG1(N297A)-HC:scFv:CD70(2H5)(C)刺激共培养物过夜，一式三份。2H5和1F6是两种不同的人CD70特异性抗体。
图21：4-1BB特异性抗体HBBK4、CD40特异性抗体G28.5和CD95特异性抗体E09的IgG1(N297)变体的scFv:CD20融合蛋白诱导与表达CD20的细胞共培养的HT1080-41BB、HT1080-CD40和HT1080(内源性表达CD95)细胞中IL8的产生。(A)融合蛋白示意图。(B-D)将所示的HT1080变体与用空载体或CD20表达质粒转染的Hek293细胞共培养。然后用指示浓度的结合CD20的IgG1(N297A)融合蛋白抗41BB(HBBK4)-IgG1(N297A)-HC:scFv:CD20(B)、抗CD40(B28.5)-IgG1(N297A)-HC:scFv:CD20(C)和抗CD95(E09)-IgG1(N297A)-HC:scFv:CD20(D)处理共培养物。第二天，通过ELISA测定IL8的产生。对于抗CD95变体，加入半胱氨酸蛋白酶抑制剂ZVAD以防止细胞凋亡。
材料与方法：HT1080-41BB转染子(Wyzgol et al.,2009)、HT1080-CD40转染子(Wyzgol et al.,2009)和HT1080细胞(内源性表达CD95)以20.000个细胞/孔的密度接种。第二天，加入20.000个已于前一天用空载体(EV)或CD27L(＝CD70)表达质粒转染的Hek293细胞。用指示浓度的抗41BB(HBBK4)-IgG1(N297A)-HC:scFvCD20(B)、抗CD40(G28.5)-IgG1(N297A)-HC:scFvCD20(C)和抗CD95(E09)-IgG1(N297A)-HC:scFvCD20(D)刺激共培养物过夜，一式三份。后者在20μg/ml ZVAD的存在下加入，以防止细胞凋亡诱导。通过ELISA测定IL8。
Wyzgol A,Müller N,Fick A,Munkel S,Grigoleit GU,Pfizenmaier K,WajantH.,Trimer stabilization,oligomerization,and antibody-mediated cell surfaceimmobilization improve the activity of soluble trimers of CD27L,CD40L,41BBL,and glucocorticoid-induced TNF receptor ligand,J Immunol,2009Aug 1；183(3):1851-61。
图22：4-1BB特异性抗体HBBK4、CD40特异性抗体G28.5和CD95特异性抗体E09的IgG1(N297)变体的scFv:CD20融合蛋白诱导与CD20阳性的BJAB细胞共培养的HT1080-41BB、HT1080-CD40和HT1080(内源性表达CD95)细胞中增强的IL8产生。(A-C)将所示的HT1080变体与CD20阳性BJAB细胞共培养，缺少内源表达CD20的Jurkat细胞作为阴性对照。然后用指示浓度的结合CD20的IgG1(N297A)融合蛋白抗41BB(HBBK4)-IgG1(N297A)-HC:scFv:CD20(A)、抗CD40(G28.5)-IgG1(N297A)-HC:scFv:CD20(B)和抗CD95(E09)-IgG1(N297A)-HC:scFv:CD20(C)处理共培养物。第二天，通过ELISA测定IL8的产生。对于抗CD95变体，加入半胱胺酸蛋白酶抑制剂ZVAD以防止细胞凋亡。
材料与方法：HT1080-41BB转染子(Wyzgol et al.,2009)、HT1080-CD40转染子(Wyzgol et al.,2009)和HT1080细胞(内源性表达CD95)以20.000个细胞/孔的密度接种。第二天，加入BJAB细胞(CD20阳性)或20.000个Jurkat细胞(CD20阴性)。用指示浓度的抗41BB(HBBK4)-IgG1(N297A)-HC:scFvCD20(A)、抗CD40(G28.5)-IgG1(N297A)-HC:scFvCD20(B)和抗CD95(E09)-IgG1(N297A)-HC:scFvCD20(C)刺激共培养物过夜，一式三份。后者在20μg/ml ZVAD的存在下加入，以防止细胞凋亡诱导。通过ELISA测定IL8。
Wyzgol A,Müller N,Fick A,Munkel S,Grigoleit GU,Pfizenmaier K,WajantH.,Trimer stabilization,oligomerization,and antibody-mediated cell surfaceimmobilization improve the activity of soluble trimers of CD27L,CD40L,41BBL,and glucocorticoid-induced TNF receptor ligand,J Immunol,2009Aug 1；183(3):1851-61。
图23：TRAILR2特异性抗体Conatumumab(Cona)的IgG1(N297A)同种型与CD70特异性scFv 9G2的融合蛋白以依赖CD70的方式增强刺激促炎性(A)和细胞毒性(B)TRAILR2信号转导。用编码CD70的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与TRAILR2反应性HT1080细胞(无CD70表达)共培养。为使HT1080细胞对TRAILR2信号敏感，用2.5μg/ml CHX(B)或CHX加20μM半胱氨酸抑制剂ZVAD(A)处理共培养物。CHX抑制具有高周转率的TRAILR2信号抑制剂FLIP蛋白的表达。ZVAD可以防止细胞凋亡，从而可以评价TRAILR2的基因诱导特性。然后用指示浓度的抗TRAILR2(Cona)-IgG1(N297A)-HC:scFv:CD70(9G2)进一步处理共培养物，再过一天后，通过ELISA(A)测定IL8的产生或通过结晶紫染色(B)测定细胞活力。
图24：IgG1(N297A)或Fab2形式的CD40特异性抗体C与衍生自靶向PD-L1的检查点抑制剂Avelumab(Ave)的scFv的融合蛋白以PD-1L限制性方式激活CD40。将编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与CD40反应性HT1080-CD40细胞共培养。然后用指示浓度的抗CD40(C)-IgG1(N297A)-HC:scFv:PD-L1(A)或抗CD40(C)-Fab2-HC:scFv:PD-L1(B)处理共培养物。第二天，通过ELISA测定IL8的产生，作为CD40激活的读数。
图25：IgG1(N297A)或Fab2形式的41BB特异性抗体HBBK与衍生自检查点抑制剂Avelumab(Ave)的scFv的融合蛋白以PD-1L限制性方式激活41BB。用编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与41BB反应性HT1080-41BB细胞共培养。然后用指示浓度的抗41BB(HBBK)-IgG1(N297A)-HC:scFv:PD-L1(A)或抗41BB(HBBK)-Fab2-HC:scFv:PD-L1(B)处理共培养物。第二天，通过ELISA测定IL41的产生，作为41BB激活的读数。
图26：PD-1L特异性抗体Avelumab(Ave)的IgG2与41BB特异性scFv的融合蛋白显示以PD-1L限制性方式增强41BB的活化。用编码PD-1L的表达质粒瞬时转染的Hek293细胞或空载体转染的Hek293细胞与41BB反应性HT1080-41BB细胞共培养。然后用指示浓度的抗PD-L1(Ave)-IgG2-HC:scFv:41BB(HBBK)处理共培养物。第二天，通过ELISA测定IL41的产生，作为41BB激活的读数。所述实施例说明，抗TNFRSF受体抗体衍生的scFv在与识别细胞表面暴露的抗原的抗体融合后获得依赖锚定的激动剂活性。因此，TRAAFFIAA原理上与抗TNFRSF受体抗体(作为TNFRSF受体的刺激效应子结构域)和scFv(作为锚定结构域)一起作用，也以镜像形式与抗体(作为锚定结构域)和TNFRSF受体特异性scFv(作为效应子结构域)起作用。
图27：鼠CD27特异性抗体的IgG(N297A)与Fn14特异性scFv 18D1的融合蛋白显示出以鼠CD27限制性方式增强的Fn14活化。将编码鼠CD27的表达质粒瞬时转染的Hek293细胞(对Fn14响应差)或空载体瞬时转染的Hek293细胞与高度Fn14响应的HT1080细胞共培养。然后用指示浓度的抗muCD27-IgG1(N297A)-HC:scFv14(18D1)处理共培养物。第二天，通过ELISA测定IL8的产生，作为Fn14激活的读数。所述实施例再次说明，抗TNFRSF受体抗体衍生的scFv在与识别细胞表面暴露的抗原的抗体融合后获得依赖锚定的激动剂活性。因此，TRAAFFIAA原理上与抗TNFRSF受体抗体(作为TNFRSF受体的刺激效应子结构域)和scFv(作为锚定结构域)一起作用，也以镜像形式与抗体(作为锚定结构域)和TNFRSF受体特异性scFv(作为效应子结构域)起作用。
发明详述
定义和通用技术
除非下文另外定义，否则应根据本领域技术人员已知的通用含义来理解本发明中使用的术语。
本文引用的所有出版物、专利和专利申请出于所有目的通过引用整体并入本文。
本文所使用的“scTNFSF”是指单链形式的功能性(意味着受体结合)TNFSF配体。通常，一个scTNFSF包含三个单元的TNFSF原聚体的受体结合结构域，其中所述单元通过多肽接头连接并形成分子内三聚体。
本文所使用的术语“抗体”是指能特异性结合目的抗原的任何功能性抗体，如Paul,W.E,(Ed.).:Fundamental Immunology 2nd Ed.,Raven Press,Ltd.,New York,1989的第7章中概述，其通过引用并入本文。在没有特别限制的情况下，术语“抗体”包括来自任何适当来源物种的抗体，包括鸡和哺乳动物，例如鼠、山羊、非人灵长类动物和人。优选地，所述抗体是人源化抗体。所述抗体优选为可以通过本领域公知的方法制备的单克隆抗体。术语“抗体”包括IgG-1、-2、-3或-4、IgE、IgA、IgM或IgD同种型抗体。术语“抗体”包括单体抗体(例如IgD、IgE、IgG)或寡聚抗体(例如IgA或IgM)。术语“抗体”还包括但不限于分离的抗体和修饰的抗体，比如基因工程抗体，例如嵌合或人源化抗体。
所述抗体结构域的命名遵循本领域已知的术语。如本领域公知的，抗体的每个单体包含两个重链和两个轻链。其中，每条重链和轻链均包含可变结构域(对于重链而言称为VH，对于轻链而言称为VL)，这对于抗原结合而言是重要的。这些重链和轻链可变结构域包含(以N末端至C末端的顺序)区域FR1、CDR1、FR2、CDR2、FR3、CDR3和FR4(FR，构架区；CDR，互补决定区，也称为高变区)。抗体序列内上述抗体区域的鉴定和分配通常根据Kabat et al.(Sequences of proteins of immunological interest,U.S.Dept.of Health and HumanServices,Public Health Service,National Institutes of Health,Bethesda,Md.1983)或Chothia et al.(Conformations of immunoglobulin hypervariableregions,Nature,1989 Dec 21-28；342(6252):877-83.)的方法，或者可以通过使用在Giudicelli et al.(IMGT/V-QUEST,an integrated software program forimmunoglobulin and T cell receptor V-J and V-D-J rearrangement analysis,Nucleic Acids Res.2004 Jul 1；32(Web Server issue):W435-40)-在此通过引用并入本文-中所述的IMGT/V-QUEST软件来进行。优选地，通过使用IMGT/V-QUEST软件鉴定和分配上述抗体区域。
“单克隆抗体”是来自基本上同质的抗体群的抗体，其中所述抗体在序列上基本上相同(即，除了含有天然存在的序列修饰例如在其N-和C-末端的氨基酸修饰的抗体的小部分不同以外)。与包含针对单个表位或针对许多不同表位的不同抗体的混合物的多克隆抗体不同，单克隆抗体针对同一表位，因此具有高度特异性。术语“单克隆抗体”包括(但不限于)从衍生自单细胞克隆的单克隆细胞群体获得的抗体，例如通过
“人源化抗体”是包含人序列和赋予与目标抗原结合特异性的一小部分非人序列的抗体。通常，人源化抗体是通过用与目标抗原结合的非人供体抗体(例如小鼠、兔、大鼠供体抗体)的高变区序列替换人受体抗体的高变区序列而产生的。在一些情况下，受体抗体的构架区序列也可以被供体抗体的相应序列替换。除了衍生自供体和受体抗体的序列外，“人源化抗体”可以包含其他(附加的或替代的)残基或序列，也可以不包含。此类其他残基或序列可用于进一步改善抗体特性，例如结合特性(例如降低Kd值)和/或免疫原性特性(例如降低人的抗原性)。产生人源化抗体的方法的非限制性实例在本领域是已知的，例如来自Riechmann et al.(Nature,1988 Mar 24；332(6162):323-7)或Jones et al.(Nature,1986 May 29-Jn 4；321(6069):522-5)。
术语“人抗体”是指包含人可变和恒定结构域序列的抗体。该定义涵盖具有单个氨基酸取代或修饰的人序列的抗体，其可用于进一步改善抗体特性，例如结合特性(例如降低Kd值)和/或免疫原性特性(例如降低人类的抗原性)。术语“人抗体”不包括赋予与目的抗原结合特异性的部分非人序列的人源化抗体。
本文所使用的抗体的“抗原结合部分”是指保留抗体特异性结合抗原的能力的抗体部分。所述能力可以，例如通过本领域已知的方法，通过确定抗原结合部分与抗体抗原特异性结合的能力来确定。抗原结合部分可以包含抗体的一个或多个片段。没有特别限制，所述抗原结合部分可以通过本领域已知的任何合适方法来产生，包括重组DNA方法和通过抗体的化学或酶促片段制备。抗原结合部分可以是Fab片段、F(ab')片段、Fab2片段、单链抗体(scFv)、单结构域抗体、双抗体或抗体的其他任何保留抗体抗感染能力的部分与抗原特异性结合。应当理解，根据本领域中已知的术语“Fab2”的含义，术语“Fab2”与术语“Fab2”、“Fab2”和“FAB2”同义。因此，出于本申请的目的，术语“Fab2”与术语“Fab2”、“Fab2”和“FAB2”可互换使用。根据本发明的“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”没有特别限制。例如，它可以是能结合肿瘤抗原的抗原结合部分。所述“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”优选选自抗CD20抗原结合部分、抗CD70抗原结合部分、抗CD19抗原结合部分、抗EGFR抗原结合部分、抗Her2抗原结合部分、抗Fn14抗原结合部分、抗CD40L抗原结合部分和抗PD1L抗原结合部分。可选地，根据本发明的“能结合细胞表面的所述结构和/或胞外基质的所述结构的抗原结合部分”可以是抗FAP抗原结合部分、抗BCMA抗原结合部分或者抗Flt3抗原结合部分。
“抗体”(例如单克隆抗体)或者“抗原结合部分”可能已被衍生化或连接至不同的分子。例如，可以与抗体连接的分子是其他蛋白质(例如其他抗体)、分子标记(例如荧光、发光、有色或放射性分子)、药剂。所述抗体或抗原结合部分可以直接连接(例如以两种蛋白质之间的融合形式)，或者通过接头分子(例如本领域已知的任何合适类型的化学接头)连接。
有关包括本发明所述融合蛋白的本发明，“抗TNFRSF受体抗体或其抗原结合部分”的含义没有特别限制，只要它是针对至少一种TNFRSF受体的抗体或其抗原结合部分即可。在根据本发明的所有其他实施方案的优选实施方案中，所述抗TNFRSF受体抗体或其抗原结合部分可以是针对单一类型的TNFRSF受体的抗TNFRSF受体抗体或其抗原结合部分。在根据本发明所有其他实施方案的另一个优选实施方案中，所述抗TNFRSF受体抗体或其抗原结合部分可以是针对两种不同类型的TNFRSF受体的二价抗TNFRSF受体抗体或其抗原结合部分。
本文所使用的术语“结合”是指与目的抗原的特异性结合。优选地，所述Kd值小于100nM，更优选地小于50nM，进一步优选地小于10nM，更进一步优选地小于5nM，最优选地小于2nM。
本文所使用的术语“表位”是指抗原的一小部分，其形成抗体的结合位点。
在本发明的上下文中，出于表征抗体或抗体融合蛋白的结合特性的目的，本发明的抗体或者所述融合蛋白(例如TRAAFFIAA)任何结合或者竞争性结合目的抗原或者FcγR或者所述细胞表面或所述胞外基质的所述结构，优选使用荧光素酶标记的(例如Gaussiaprinceps luciferase(GpL))抗体或融合蛋白的变体，通过细胞结合研究(例如Kums etal.,MAbs,2017Apr；9(3):506-520中所述)或者表面等振离子共振测量作为参考标准测定。
术语“KD”或“KD值”是指本领域已知的平衡解离常数。在本发明的上下文中，这些术语是指本发明的抗体或融合蛋白(例如TRAAFFIAA)相对于具体的目的抗原或者目的FcγR或者相对于细胞表面或胞外基质的所述结构的平衡解离常数。所述平衡解离常数是复合物(例如抗原-抗体复合物)可逆地解离为其组分(例如抗原和抗体)的倾向的量度。对于根据本发明的抗体或融合蛋白，KD值优选通过GpL标记的所述抗体或融合蛋白(例如TRAAFFIAA)的变体与未修饰的目的抗体或融合蛋白(例如TRAAFFIAA)的胞内竞争性结合研究或表面等振离子共振测定。
本文所使用的结合本发明的所述抗体或其抗原结合部分，术语“能与抗体交叉竞争”以与具体蛋白质例如TNFRSF受体结合通常是指在使用本领域已知的分析方法，例如酶联免疫吸附测定法(ELISA)时，抗体或其抗原结合部分能交叉竞争。如本领域已知的，应当理解的是，术语“能与抗体交叉竞争”是指与所述具体蛋白质特异性结合的交叉竞争。本领域技术人员将能确定用于检测这种特异性结合的交叉竞争的合适条件。
本文所使用的“分离的抗体”或“分离的TRAAFFIA”或“分离的融合蛋白”已被鉴定并与其来源环境的大部分组分(按重量计)分离，例如，来自杂交瘤细胞培养物或用于生产的不同细胞培养物的组分(例如重组表达抗体或融合蛋白(例如TRAAFFIA)的生产细胞，例如CHO或HEK293细胞)。进行所述分离是为了充分去除可能干扰所述抗体或融合蛋白(如TRAAFFIA)适用所需用途的组分(例如，根据本发明的所述抗体或融合蛋白(如TRAAFFIA)的治疗用途)。制备分离的抗体或抗体融合蛋白的方法是本领域已知的，包括蛋白A色谱法、阴离子交换色谱法、阳离子交换色谱法、病毒保留过滤和超滤。优选地，通过Lowry蛋白测定法测定，所述分离的抗体或融合蛋白(如TRAAFFIA)制剂至少为70％纯(w/w)，更优选地至少为80％纯(w/w)，进一步优选地为至少90％纯(w/w)，更进一步优选地至少95％纯(w/w)，最优选地为至少99％纯(w/w)。
本文所使用的“双抗体”是二价抗原结合抗体的一小部分，其包含重链可变结构域，所述重链可变结构域通过肽接头连接至同一多肽链上的轻链可变结构域，所述肽接头太短而无法使同一条链上的两个结构域配对。这导致与另一条链的互补结构域配对，并导致具有两个抗原结合位点的二聚体分子组装。双价抗体可以是双价和单特异性的(例如具有两个同一抗原的结合位点的双抗体)，也可以是双价和双特异性的(例如具有两个抗原结合位点的双价抗体，一个是抗原的结合位点，另一个是另一抗原的结合位点)。双抗体的详细描述可以在Holliger P et al.(““Diabodies”:small bivalent and bispecificantibody fragments.”,Proc Natl Acad Sci U S A.1993 Jul 15；90(14):6444-8)中找到。
本文所使用的“单结构域抗体”(也称为“纳米抗体TM”)是由单个单体可变抗体结构域组成的抗体片段。产生单结构域抗体的结构和方法是本领域已知的，例如Holt LJ etal.(”Domain antibodies:proteins for therapy.”Trends Biotechnol.2003 Nov；21(11):484-90)、Saerens D et al.(”Single-domain antibodies as building blocksfor novel therapeutics.”Curr Opin Pharmacol.2008 Oct；8(5):600-8.Epub 2008 Aug22)，和Arbabi Ghahroudi M et al.(“Selection and identification of singledomain antibody fragments from camel heavy-chain antibodies.”FEBS Lett.1997Sep 15；414(3):521-6)。
在本文中与本发明有关的“融合蛋白”不限于特定类型的融合蛋白，只要融合蛋白的各部分通过共价键融合即可。例如，所述融合蛋白的各部分可以通过在一条或多条单多肽链中表达、通过一个或多个二硫键连接、通过化学偶联(优选通过使用点击化学的化学偶联)和/或通过任何其他本领域已知的适合蛋白质连接的共价连接而融合。优选地，所述融合蛋白的各部分通过在一条或多条单多肽链中的表达和/或通过一个或多个二硫键而融合。因此，在本发明的融合蛋白中，抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域，可通过在一条或多条单多肽链中表达、通过一个或多个二硫键连接、通过化学偶联(优选使用点击化学的化学偶联)和/或通过任何其他本领域已知的适合蛋白质连接的共价连接而融合。优选地，所述抗TNFRSF受体抗体或其抗原结合部分与能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域，通过在一条或多条单多肽链中表达和/或通过一个或多个二硫键连接而融合。在本发明的所有实施方案中，其中，能以不依赖FcγR的方式与细胞表面的结构和/或胞外基质的结构结合的结构域包括：
-抗体的抗原结合部分，所述抗原结合部分能与细胞表面的所述结构和/或胞外基质的所述结构结合，其中能与细胞表面和/或胞外基质的所述结构结合的所述抗原结合部分是与一种或多种FcγR类型结合的能力降低的抗原结合部分，所述抗原结合部分是Fab2片段；或
-能结合细胞表面的所述结构和/或胞外基质的所述结构的全长抗体；
非常优选地，所述抗TNFRSF受体抗体或其抗原结合部分与所述结构域通过化学偶联，优选使用点击化学的化学偶联来融合。
通常，就本发明的所有融合蛋白而言，应当理解的是，所述融合蛋白的各部分(例如抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域)可以通过接头序列融合。在这种情况下，本发明的融合蛋白将包含此类接头序列。合适的接头序列是本领域已知的，并且包括例如肽接头，但不限于此。例如，在本发明所述的通过在一条或多条单多肽链中表达而融合的抗TNFRSF受体抗体或其抗原结合部分以及能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域的融合蛋白中，所述一条或多条单多肽链可包含一个或多个肽接头序列，所述肽接头序列位于所述抗TNFRSF受体抗体或其抗原结合部分与能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域之间。
就本发明而言，术语“能以不依赖FcγR的方式结合所述细胞表面的结构或者所述胞外基质的结构的结构域”是指所述结构域能与不同于FcγR的所述细胞表面的结构和/或所述胞外基质的结构结合。除非另有说明，否则不排除所述结构域还可能包含能与FcγR结合的其他部分(例如Fc结构域)的可能性。
根据本发明，术语“包含”的每次出现可以可选地被术语“由...组成”代替。
方法与技术
通常，除非本文另有定义，否则本发明中使用的方法(例如，克隆方法或与抗体有关的方法)是根据本领域已知的方法进行的，例如，Sambrook et al.(“MolecularCloning:A Laboratory Manual.”,2nd Ed.,Cold Spring Harbor Laboratory Press,ColdSpring Harbor,New York 1989)、Ausubel et al.(“Current Protocols in MolecularBiology.”Greene Publishing Associates and Wiley Interscience；New York 1992)和Harlow and Lane(“Antibodies:A Laboratory Manual”,Cold Spring HarborLaboratory Press,Cold Spring Harbor,New York 1988)中所述的方法，所有其通过引用并入本文。
融合蛋白及其结构域与它们各自的靶蛋白的结合可以通过本领域已知的方法来评估。所述结合优选通过表面等振离子共振测定来评估。
根据本发明，对序列通过使用BLAST算法进行序列比对(参见Altschul et al.(1990)”Basic local alignment search tool.”Journal of Molecular Biology215.p.403-410.；Altschul et al.:(1997)Gapped BLAST and PSI-BLAST:a newgeneration of protein database search programs.NucleicAcids Res.25:3389-3402，其全部通过引用并入本文)。优选地，使用以下参数：最大靶序列10；和字长3；BLOSUM 62矩阵；空位罚分：存在空位11分，延伸1分；条件成分分数矩阵调整。因此，当与序列结合使用时，诸如“同一性”或“相同性”的术语是指通过使用BLAST算法获得的同一性值。
根据本发明，抗体融合蛋白可以通过本领域已知的任何方法产生，包括但不限于在Siegel DL(“Recombinant monoclonal antibody technology.”TransfusClinBiol.2002Jan；9(1):15-22，其通过引用并入本文)中提及的方法。
TNFRSF受体的刺激可以通过本领域已知的任何方法测定。例如，在TNFRSF受体为TNFR2、CD40、CD95、Fn14的情况下，可以通过使用基于细胞的IL8分泌测定法、并随后通过ELISA检测分泌的IL8。可用于此类基于细胞的IL8分泌测定的细胞的非限制性示例为HT1080 CD40转染子(可用于测定CD40刺激)、Widr细胞(可用于测定Fn14刺激)、H1080-细胞(可用于测定CD95刺激)，以及表达TNFR2的HeLa转染子(可用于测定TNFR2刺激)。此类测定的非限制性实施例可以在实施例1-3和相应的图中找到。对于当刺激时具有细胞毒性作用的TNFRSF受体，例如CD95，优选将细胞可渗透的半胱氨酸蛋白酶抑制剂例如ZVAD添加到测定中，以防止诱导细胞死亡。本发明的融合蛋白(例如TRAAFFIAAS)对各种TNFRSF受体的刺激也可以通过任何能检测经典NFκB途径的刺激(例如IκBα的磷酸化和降解、IKK激活、p65的磷酸化和核易位、检测靶基因)或NFκB旁路途径的刺激(例如p100降解、NIK积累)。TRAAFFIAA对CD95和TRAIL死亡受体(DR4、DR5)的刺激作用也可以通过各种测定细胞死亡诱导方法来评估。TRAAFFIA对CD40的刺激可进一步通过测定树突细胞的成熟度来评估。
本发明组合物的制备
根据用于制备药物组合物的已知标准来制备根据本发明的组合物。
例如，所述组合物以其可被适当地储存和施用的方式制备，例如，通过添加药学上可接受的组分，例如载体、赋形剂或稳定剂。
当向患者施用所述药物组合物时，这样的药学上可接受的组分就使用量而言是无毒的。添加到药物组合物中的药学上可接受的组分可以取决于所述药物组合物的具体预期用途和给药途径。
通常，用于本发明的药学上可接受的组分是根据本领域可获得的知识来使用的，例如来自Remington's Pharmaceutical Sciences，Ed.AR Gennaro,20th edition,2000,Williams&Wilkins,PA,USA。
序列
本申请使用的氨基酸序列以单字母氨基酸编码按照N-末端至C-末端的顺序表示。本申请使用的核苷酸序列以标准核酸代码按照5'至3'的顺序表示。星号表示蛋白质序列的末端。带下划线的核酸序列代表限制性核酸内切酶位点。
以下非限制性示例性序列用于本申请的实施例中：
表1:
SEQ ID NO:1:抗CD40-Flag-VH-heavy-full-scTNF80(mu)-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcgagttcacacgggacaaacctgtggctcatgtggtggccaatcatcaggtggaggaacagctggaatggctgagtcagagagcaaacgccctgctggcaaatgggatggacctcaaagacaatcagctcgtggtgcctgccgatggactgtacctggtgtactctcaggtcctgtttaagggacagggatgccccgattacgtgctgctcacccacactgtgtcacgcttcgccatctcataccaggagaaagtcaatctcctctccgccgtgaaatcaccatgtcctaaggatactcccgagggagccgaactgaaaccttggtacgaacccatctacctgggcggcgtgtttcagctggagaaaggcgatcagctctccgccgaagtgaatctgcccaaatacctcaactttagggaatccggacaggtctactttggcgtgattgccctgggaggcggatctggaggaggctctggcgggggatctgggggcggatccgacaaacctgtggcacacgtcgtggcaaaccatcaggtcgaggaacagctcgagtggctgtcacagagggccaatgccctgctggcaaatggaatggatctgaaggataatcagctcgtcgtgcctgccgacggcctctacctcgtctactctcaggtcctctttaagggacagggctgccccgactacgtcctgctcactcataccgtgagtcgcttcgctatttcataccaggaaaaagtcaacctgctgagtgctgtgaaatctccttgccctaaggatacccctgagggagccgaactcaaaccatggtacgagccaatctacctcggaggagtgtttcagctggaaaaaggggatcagctctccgccgaagtcaacctccccaaatacctcaatttccgggaatccggacaggtgtactttggagtcattgccctgggaggcggctctggcgggggatctggaggaggctccggaggaggcagtgacaaacccgtcgctcacgtggtggcaaatcatcaggtcgaggaacagctggaatggctgtctcagagagcaaacgctctcctcgccaatggaatggatctcaaggacaaccagctcgtcgtccctgccgatggactctacctggtctactctcaggtgctctttaagggacagggatgccccgattacgtcctgctcacacacaccgtgtctcgctttgctatttcataccaggagaaagtcaatctgctgtctgccgtcaaatctccttgtccaaaagacacacccgagggagccgaactcaaaccttggtacgagccaatctacctggggggagtgtttcagctggagaagggggatcagctctccgccgaagtgaatctcccaaaatacctcaattttcgggaatccggacaggtctactttggagtgattgccctgtag
SEQ ID NO:19:SEQ ID NO:1表达的蛋白质构建体：
SEQ ID NO:2:抗CD40-Flag-VH-heavy-full-pCR3(hIgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:20:SEQ ID NO:2表达的蛋白质构建体：
SEQ ID NO:3:抗CD40-Flag-VH-heavy-full-scFv-抗CD20-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaAatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagCcaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggcgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaGgggggaccaagctggaaatcaaacgttaa
SEQ ID NO:21:SEQ ID NO:3表达的蛋白质构建体：
SEQ ID NO:4:抗CD40-Flag-VH-heavy-full-scBaff-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:22:SEQ ID NO:4表达的蛋白质构建体：
SEQ ID NO:5:抗CD40-Flag-VL-light-full-pCR3:
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatcgtgatgactcagaacccactgtctctgcctgtgtctctgggggatgaggctagcatttcttgccgctcatctcagtcactggagaactccaatggcaacaccttcctgaattggtttttccagaaacccggccagtcacctcagctgctcatctaccgagtgagcaatcggtttagcggagtgcccgatcgattctctggctccggatctgggaccgactttaccctgaaaatctcacgagtggaggccgaggatctgggagtgtacttctgtctccaggtcacacatgtgccttacacatttggcggcggaacaactctcgaaatcaaaggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:23:SEQ ID NO:5表达的蛋白质构建体：
SEQ ID NO:6:C4-HC-full-IL2(mu)-pCR3(hIgG1)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattgcccggtaccgcccctaccagcagcagcacctctagctctacagccgaggctcaacaacaacaacaacagcaacagcagcagcagcagcacctggaacagctgctgatggacctgcaggaactgctgagccggatggaaaactaccggaacctgaagctgccccggatgctgaccttcaagttctacctgcccaagcaggccaccgagctgaaggatctgcagtgcctggaagatgagctgggccccctgagacacgtgctggatctgacccagagcaagagctttcagctggaagatgccgagaacttcatcagcaacatcagagtgaccgtcgtgaagctgaagggcagcgacaacaccttcgagtgccagttcgacgacgagagcgctaccgtggtggacttcctgcggagatggatcgccttctgccagagcatcatcagcaccagcccccagtaa
SEQ ID NO:24:SEQ ID NO:6表达的蛋白质构建体：
SEQ ID NO:7:C4-HC-heavy-konst-full-scFv-抗CD19-pCR3(N297A)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcgacattcagatgacgcagtctccatcctccatgtctgtatctctgggagacacagtcagcatcacttgccatgcaagtcagggcattagcagtaatatagggtggttgcagcagaaaccagggaaatcatttaagggcctgatctatcatggaaccaacttggaagatggagttccatcaaggttcagtggcagtggatctggagcagattattctctcaccatcagcagcctggaatctgaagattttgcagactattactgtgtacagtatgctcagtttccgtacacgttcggaggggggaccaagctggagctgaaacgtggtggtggtggttctggtggtggtggttctggcggcggcggctccagtggtggtggatcccaggttcagctgcagcaatctggacctgagctggtgaagcctggggcctcagtgaagatttcctgcaaagcttctggctacgcattcagtagctcttggatggactgggtgaagcagaggcctggacagggtcttgagtggattggacggatttatcctggagatggagatactaactacaatgggaagttcaagggcaaggccacactgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacctctgtggactctgcggtctatttctgtgcaaggtccattactacggtagtagggtggtacttcgatgtctggggcgcagggaccacggtcaccgtttcctcctaa
SEQ ID NO:25:SEQ ID NO:7表达的蛋白质构建体：
SEQ ID NO:8:C4-HC-heavy-konst-full-scFv-抗CD20-pCR3(N297A)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggaccctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:26:SEQ ID NO:8表达的蛋白质构建体：
SEQ ID NO:9:C4-HC-heavy-full-pCR3(IgG1)(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:27:SEQ ID NO:9表达的蛋白质构建体：
SEQ ID NO:10:C4-HC-heavy-full-scGITRL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcggcggagggagcggacagctggaaactgccaaagaaccctgtatggccaaattcggaccactgcctagcaaatggcagatggctagtagcgaacctccatgtgtgaacaaagtgagcgattggaaactcgagatcctccagaatggactgtacctcatctacggacaggtcgccccaaatgccaattacaatgatgtggccccctttgaagtccggctctacaaaaacaaggatatgatccagaccctcaccaacaaatccaaaatccagaatgtgggcggcacatacgaactccatgtcggcgataccatcgatctcattttcaactctgaacaccaggtgctcaaaaacaacacctactggggaatcatcctgctggcaaaccctcagttcatctccggcggcggctctggcggcggatctggcggagggagtggcggaggctcacagctggagactgctaaagaaccctgtatggcaaaattcgggcccctgccctcaaaatggcagatggcctcctctgaaccaccctgtgtgaacaaagtgagtgattggaaactggaaatcctccagaacggcctctacctcatctacggacaggtggcacccaatgccaattacaacgacgtggcacccttcgaagtgagactgtacaaaaacaaggatatgatccagaccctcaccaacaaatcaaaaatccagaatgtcggagggacatacgaactccatgtgggagacactatcgacctcattttcaattccgaacatcaggtcctgaaaaacaacacttactggggcattattctgctcgccaatccacagtttattagtggaggcgggggatctgggggaggctccggcggagggagtggaggcggatctcaattccaattagagactgctaaggagccctgtatggctaagtttggaccattaccctcaaaatggcaaatggcatcttctgaacctccttgcgtgaataaggtgtctgactggaagctggagatacttcagaatggcttatatttaatttatggccaagtggctcccaatgcaaactacaatgatgtagctccttttgaggtgcggctgtataaaaacaaagacatgatacaaactctaacaaacaaatctaaaatccaaaatgtaggagggacttatgaattgcatgttggggacaccatagacttgatattcaactctgagcatcaggttctaaaaaataatacatactggggtatcattttactagcaaatccccaattcatctcctag
SEQ ID NO:28:SEQ ID NO:10表达的蛋白质构建体：
SEQ ID NO:11:C4-HC-heavy-full-sc(mu)41BBL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcggcggtggaagtggtactgaaccgcgacccgctctcactataaccaccagccctaacttgggaacgcgggagaataacgcggatcaagtaaccccagtttcccatatcgggtgtccaaataccactcaacaaggatcaccggtctttgctaagctcctggcgaaaaaccaagcgtccctctgtaatacaacccttaattggcactcccaagatggggcgggttcttcatacctctcacaggggctccgatacgaagaagacaaaaaggagttggtggtcgactcaccaggactttactatgtattcctggagttgaagctctctcctacattcaccaataccggtcacaaagtacagggttgggtgagccttgtgctccaggcgaagccgcaggtggacgacttcgacaaccttgcactcactgtagagctgttcccttgttccatggagaataaattggtggaccgctcttggagtcagctcctcctcctcaaggcgggtcatcgattgagcgttggccttcgggcatatcttcatggggcgcaagatgcataccgcgattgggaattgtcataccccaacacgacctcttttggcctgtttctggtcaaacccgacaatccgtgggaaggtgggggaagcggagggggttcaggaggaggatctgggggaggttcaactgaaccgaggcccgcgcttactatcacgacttccccaaatctggggactagggaaaacaatgccgatcaggtcactcctgttagtcacattggttgtcctaatacgacccagcaaggctctccggtgtttgccaaactgttggccaaaaatcaggcgtcactttgtaatacaacgctcaactggcatagtcaggatggggccggctcctcatacttgtctcaaggtcttaggtacgaagaagataagaaggagctggtggtagacagccccgggctctactatgtgttcctggagctcaaactgtcaccgacgttcactaacaccggtcataaggtacagggttgggtatccttggtgttgcaagcaaaaccccaggtggacgatttcgataatcttgcgcttactgtagagctctttccatgttcaatggaaaataaactggtcgataggagctggtcccaacttctccttcttaaagctggccatcgcctgagtgttggcctgagagcgtatcttcatggggcgcaggacgcttaccgggattgggaactgtcatacccaaacaccaccagctttgggctcttccttgtaaagccagacaatccgtgggaggggggaggcgggagtgggggcgggtctggagggggcagtggggggggtagtacggagccgcgccccgccctgaccatcacaacgtcacccaatcttgggactcgggagaataacgccgaccaggttacccctgtatcccatatcggttgtcctaatacgacacaacaaggcagtcctgtattcgctaaactcttggcaaaaaaccaggccagtctttgtaatacgacgcttaattggcatagccaggacggtgcgggcagctcctacctttcccaggggctcaggtatgaagaagataagaaagaactcgttgtagacagtcccggattgtattacgtttttttggaactcaagctctctccaaccttcaccaatacgggacataaggtccagggctgggtgagcctcgtactccaggctaagccgcaagttgacgatttcgataatctcgctcttacagtggagttgtttccctgtagtatggagaataagctcgtcgaccggtcttggagccaacttctgctgcttaaggctggtcaccggctcagtgtaggcctccgagcgtatttgcatggggcgcaggacgcctatcgagactgggagctttcctaccctaacacgaccagctttggactcttcttggtgaaacctgacaatccgtgggaataa
SEQ ID NO:29:SEQ ID NO:11表达的蛋白质构建体：
SEQ ID NO:12:C4-HC-heavy-full-(mu)GITRL-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattcccaactgccatcgagtcctgcatggttaagtttgaactatcatcctcaaaatggcacatgacatctcccaaacctcactgtgtgaatacgacatctgatgggaagctgaagatactgcagagtggcacatatttaatctacggccaagtgattcctgtggataagaaatacataaaagacaatgcccccttcgtagtacagatatataaaaagaatgatgtcctacaaactctaatgaatgattttcaaatcttgcctataggaggggtttatgaactgcatgctggagataacatatatctgaagttcaactctaaagaccatattcagaaaactaacacatactgggggatcatcttaatgcctgatctaccattcatctcttag
SEQ ID NO:30:SEQ ID NO:12表达的蛋白质构建体：
SEQ ID NO:13:C4-LC-light-full-pCR3(无Flag):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtggatactgctgtagcctggtatcaacaaaaaccagggcaatctcctaaactactgatttactgggcatccacccggcacactggagtccctgatcgcttcacaggcagtggatctgggacagattatactctcaccatcagcagtgtgcaggctgaagacctggcgcgttattactgtcagcaatattatagtgttcctccgacgttcggtggaggcaccaagctgggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:31:SEQ ID NO:13表达的蛋白质构建体：
SEQ ID NO:14:抗CD95(E09)-Flag-VH-full-scFv-抗CD19-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggacattcagatgacgcagtctccatcctccatgtctgtatctctgggagacacagtcagcatcacttgccatgcaagtcagggcattagcagtaatatagggtggttgcagcagaaaccagggaaatcatttaagggcctgatctatcatggaaccaacttggaagatggagttccatcaaggttcagtggcagtggatctggagcagattattctctcaccatcagcagcctggaatctgaagattttgcagactattactgtgtacagtatgctcagtttccgtacacgttcggaggggggaccaagctggagctgaaacgtggtggtggtggttctggtggtggtggttctggcggcggcggctccagtggtggtggatcccaggttcagctgcagcaatctggacctgagctggtgaagcctggggcctcagtgaagatttcctgcaaagcttctggctacgcattcagtagctcttggatggactgggtgaagcagaggcctggacagggtcttgagtggattggacggatttatcctggagatggagatactaactacaatgggaagttcaagggcaaggccacactgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacctctgtggactctgcggtctatttctgtgcaaggtccattactacggtagtagggtggtacttcgatgtctggggcgcagggaccacggtcaccgtttcctcctaa
SEQ ID NO:32:SEQ ID NO:14表达的蛋白质构建体：
SEQ ID NO:15:抗CD95-E09-Flag-VH-heavy-full-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccctgaactgctgggcggaccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcccagagaggaacagtacaacagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcctcccagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctacccctccgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagtaa
SEQ ID NO:33:SEQ ID NO:15表达的蛋白质构建体：
SEQ ID NO:16:抗CD95(E09)-Flag-VH-FAB2(1-114)-scFv-抗CD20-pCR3(IgG1):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagcaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:34:SEQ ID NO:16表达的蛋白质构建体：
SEQ ID NO:17:抗CD95-E09-Flag-VH-heavy-full-scBaff-pCR3(N297A):
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:35:SEQ ID NO:17表达的蛋白质构建体：
SEQ ID NO:18:抗CD95-E09-Flag-VL-light-full-pCR3:
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagagcgtgctgacccagcctcctagcgtgtccgaagcccctagacagaccgtgaccatcagctgctccggcaacagcttcaacatcggcagataccccgtgaactggtatcagcagctgccaggcaaggcccctaaactgctgatctactataacaacctgcggttcagcggagtgtccgaccggttctctggcagcaagtctggcacatctgccagcctggccatccgggatctgctgtctgaggacgaggccgactactactgcagcacctgggacgacaccctgaagggctgggtgttcggcggaggcaccaaagtgacagtgctgggcg gatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:36:SEQ ID NO:18表达的蛋白质构建体：
表2：CDR序列(根据Kabat)。注意，所述重链和轻链CDR序列分别由“-H”和“-L”指示。
SEQ ID NO:96:SEQ ID NO:21所示蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:97:SEQ ID NO:24所示的蛋白质构建体的鼠源IL-2锚定结构域：
SEQ ID NO:98:SEQ ID NO:25所示的蛋白质构建体的scFv-抗CD19锚定结构域：
SEQ ID NO:99:SEQ ID NO:26所示的蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:100:SEQ ID NO:32所示的蛋白质构建体的scFv-抗CD19锚定结构域：
SEQ ID NO:101:SEQ ID NO:34所示的蛋白质构建体的scFv-抗CD20锚定结构域：
SEQ ID NO:102:SEQ ID NO:19所示的蛋白质构建体的scTNF80(鼠)锚定结构域：
SEQ ID NO:103:SEQ ID NO:22所示的蛋白质构建体的scBaff锚定结构域：
SEQ ID NO:104:SEQ ID NO:28所示的蛋白质构建体的scGITRL锚定结构域：
SEQ ID NO:105:SEQ ID NO:29所示的蛋白质构建体的鼠sc41BBL锚定结构域：
SEQ ID NO:106:SEQ ID NO:30所示的蛋白质构建体的鼠GITRL锚定结构域：
SEQ ID NO:107:人IL-2锚定结构域：
SEQ ID NO:108:抗CD40(G28.5)-VH(1-114)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcctaa
SEQ ID NO:117:SEQ ID NO:108所示的蛋白质构建体：
SEQ ID NO:109:抗CD40(G28.5)-VH(1-114)-scBaff
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggatatccagctccagcagtctggccctggactcgtcaaaccatctcagagcctgtctctcacctgttctgtcaccggatactccatcaccaccaactacaactggaattggattcggcagtttcctgggaacaaactcgaatggatgggatacatccgatacgacggcactagtgaatacaccccatctctcaaaaatcgggtgtccattacccgggacacttctatgaaccagttctttctccgactcacctctgtgacacctgaggataccgccacatactactgtgctagactggactactgggggcagggaacactggtgaccgtgtcatctggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:118:SEQ ID NO:109所示的蛋白质构建体
SEQ ID NO:110:抗CD95(E09)-VH(1-114)-scBaff
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgagggacccgaggaaactgtgactcaggactgtctccagctcattgccgatagtgaaacccctaccatccagaaaggctcttacaccttcgtgccatggctgctgtcattcaaacggggatctgctctggaggagaaggaaaacaaaatcctggtcaaggaaaccggctacttcttcatctacggccaggtcctctacaccgacaaaacatacgctatggggcatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtattcagaacatgcccgagactctgcccaataatagctgctactctgctggcattgccaaactggaggagggcgacgaactccagctggctattcctagggaaaatgcccagattagcctggacggggatgtgacattttttggcgccctgaaactgctgggaggcggagggagtggcgggggaggctctggacctgaggaaactgtgacccaggattgtctccagctcattgccgatagtgagactcctaccattcagaagggatcttacacctttgtgccttggctgctgtctttcaaacggggctctgctctggaggaaaaggagaacaaaattctggtcaaagagactggctacttcttcatctacggccaggtgctgtacaccgacaaaacatacgccatgggccatctcattcagcggaaaaaagtccacgtgttcggcgacgaactctctctcgtgacactgttccggtgtatccagaacatgcccgagacactgcccaataatagctgctactctgccggcattgctaaactggaggagggggacgaactccagctggctattcctagggaaaatgcccagatttctctcgatggggatgtgacattcttcggggccctcaaactgctgggaggcggcggatctggcggaggcgggagtcaattcgcagcaggtccagaagaaacagtcactcaagactgcttgcaactgattgcagacagtgaaacaccaactatacaaaaaggatcttacacatttgttccatggcttctcagctttaaaaggggaagtgccctagaagaaaaagagaataaaatattggtcaaagaaactggttacttttttatatatggtcaggttttatatactgataagacctacgccatgggacatctaattcagaggaagaaggtccatgtctttggggatgaattgagtctggtgactttgtttcgatgtattcaaaatatgcctgaaacactacccaataattcctgctattcagctggcattgcaaaactggaagaaggagatgaactccaacttgcaataccaagagaaaatgcacaaatatcactggatggagatgtcacattttttggtgcattgaaactgctgtga
SEQ ID NO:119:SEQ ID NO:110所示的蛋白质构建体
SEQ ID NO:111:抗TNFR2(C4)-IgG2-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatcctcgagtgctagcaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcggcgtgcacaccttcccagctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaagaatt ccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:120:SEQ ID NO:111所示的蛋白质构建体
SEQ ID NO:112:抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(1F6)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccagattcagctcgtccagtccggacctgaagtgaaaaaacctggcgaaaccgtgaaaatttcctgtaaggcctctggctacacctttaccaactacggcatgaactgggtcaaacaggctcctgggaagggcctgaaatggatgggatggatcaacacctacaccggcgaaccaacatacgccgatgcctttaagggacgctttgccttctctctggaaacttccgcctctactgcttacctccagatcaataacctcaaaaacgaggacaccgccacttacttttgtgctcgggattacggggactacgggatggattactggggacagggaacatctgtgaccgtgtctagcgcttctacaaaggggcctaaactggaggagggcgagtttagcgaggctagagtggatatcgtgctcacacagtctcccgcttctctggctgtctcactgggccagcgagcaacaatctcttgtcgggcttccaaatccgtgtctactagcggctactcttttatgcactggtaccagcagaaacctgggcagcctccaaaactgctcatctacctggcttcaaacctcgaatccggagtgcctgctcgattttctggctctggctccgggaccgactttacactgaacattcatcctgtcgaggaggaggacgctgccacatactactgtcagcattctagggaggtgccatggacatttggcgggggaacaaaactggaaatcaaacggtaa
SEQ ID NO:121:SEQ ID NO:112所示的蛋白质构建体：
SEQ ID NO:113:抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(2H5)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattgcaggttcagctgctgcagtctggacctgagctggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggttatagtttcacaagttacgatattaactgggtgaagcagaggcctggacagggacttgagtgggttggatggatttatcctagagatggtgatactaagtacaatgagaaattcaagggcaaggccatattgactgtagacacatcctccaacacagcgtacatgaacctccacagcctgacatctgaggactctgcggtctatttctgtgcaagactaactgggccctactggtacttcgatgtctggggcacagggaccacggtcaccgtctcctcaggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtgcagctggtggaatctggcggcggagtggtgcagcctggcagaagcctgagactgagctgtgccgccagcggcttcaccttcagcagctacatcatgcactgggtgcgccaggcccctggcaagggactggaatgggtggccgtgatcagctacgacggccggaacaagtactacgccgacagcgtgaagggccggttcaccatctcccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgcgggccgaggacaccgccgtgtactactgtgccagagacaccgacggctacgacttcgactattggggccagggcaccctcgtgaccgtgtctagcggaggcggaggatctggcggagggggatcaggcgggggaggctctgaaatcgtgctgacacagagccccgccaccctgtcactgtctccaggcgaaagagccaccctgagctgcagagccagccagagcgtgtccagctacctggcctggtatcagcagaagcccggacaggcccccagactgctgatctacgacgccagcaatcgggccacaggcatccctgccagattttccggctctggcagcggcaccgacttcaccctgacaatcagcagcctggaacccgaggactttgccgtgtattattgccagcagcggaccaactggcccctgacctttggcggaggcaccaaggtggaaatcaaggccagcaccaagggctaa
SEQ ID NO:122:SEQ ID NO:113所示的蛋白质构建体：
SEQ ID NO:114:抗41BB(HBBK4)-IgG1(N297A)-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgagcaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcctgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcggatccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:123:SEQ ID NO:114所示的蛋白质构建体：
SEQ ID NO:115:抗41BB(HBBK4)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatcgtgatgactcagagccccgccttcctgtctgtgacccctggcgagaaagtgaccatcacctgtagagccagccagaccatcagcgactacctgcactggtatcagcagaagcccgatcaggcccctaagctgctgattaagtacgcctctcagagcatcagcggcatccccagcagattttctggcagcggctctggcaccgacttcacctttaccatcagctccctggaagccgaggatgccgccacctactattgtcaggacggccacagcttccctccaacctttggacagggcacaaagctggaaatcaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:124:SEQ ID NO:115所示的蛋白质构建体：
SEQ ID NO:116:抗CD95(E09)-IgG1(N297A)-VH-scFv:CD20
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagctgcagctgcaggaatctggccctggcctcgtgaagcccagcgagacactgagcctgacctgtaccgtgtccggcgccagcatcagcgccaacagctactatggcgtgtgggtgcgccagagccctggcaagggactggaatgggtgggatctatcgcctaccggggcaacagcaacagcggcagcacctactacaaccccagcctgaagtcccgggccaccgtgtctgtggacaccagcaagaaccaggtgtccctgcggctgacctctgtgacagccgccgataccgccctgtactactgcgccagaaggcagctgctggacgacggcacaggatatcagtgggccgccttcgatgtgtggggccagggaacaatggtcaccgtgtcctccggatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaaggaattccaggtacaactgcagcagcctggggctgagctggtgaagcctggggcctcagtgaagatgtcntgcaaggcttctggctacacatttaccagttacaatatgcactgggtaaaacagacacctggtcggggcctggaatggattggagctatttatcccggaaatggtgatacttcctacaatcagaagttcaaaggcaaggccacattgactgcagacaaatcctccagcacagcctacatgcagctcagcagcctgacatctgaggactctgcggtctattactgtgcaagatcgacttactacggcggtgactggtacttcaatgtctggggcgcagggaccacggtcaccgtctcttcaggaggaggcgg atccggcggaggcggaagcggtggcggaggctctcaaattgttctctcccagtctccagcaatcctgtctgcatctccaggggagaaggtcacaatgacttgcagggccagctcaagtgtaagttacatccactggttccagcagaagccaggatcctcccccaaaccctggatttatgccacatccaacctggcttctggagtccctgttcgcttcagtggcagtgggtctgggacttcttactctctcacaatcagcagagtggaggctgaagatgctgccacttattactgccagcagtggactagtaacccacccacgttcggaggggggaccaagctggaaatcaaacgttaa
SEQ ID NO:125:SEQ ID NO:116所示的蛋白质构建体
以下如SEQ ID NO 126至143的氨基酸序列是抗4-1BB单克隆抗体HBBK4(其在本申请的非限制性实验中使用)、URELUMAB和UTOMILUMB的CDR序列。注意，术语“CDR1-H”、“CDR2-H”和“CDR3-H”分别指重链CDR1、CDR2和CDR3序列，术语“CDR1-L”、“CDR2-L”和“CDR3-L”分别指轻链CDR1、CDR2和CDR3序列：
SEQ ID NO:126:HBBK4 CDR1-H:
YTFSSYWMH
SEQ ID NO:127:HBBK4 CDR2-H:
EINPGNGHTNYNEKFKS
SEQ ID NO:128:HBBK4 CDR3-H:
SFTTARAFAY
SEQ ID NO:129:HBBK4 CDR1-L:
RASQTISDYLH
SEQ ID NO:130:HBBK4 CDR2-L:
LASQSIS
SEQ ID NO:131:HBBK4 CDR3-L:
QDGHSFPPT
SEQ ID NO:132:URELUMAB CDR1-H:
GSFSGYYWS
SEQ ID NO:133:URELUMAB CDR2-H:
EINHGGYVTYNPSLES
SEQ ID NO:134:URELUMAB CDR3-H:
DYGPGNYDWYFDL
SEQ ID NO:135:URELUMAB CDR1-L:
RASQSVSSYLA
SEQ ID NO:136:URELUMAB CDR2-L:
DASNRAT
SEQ ID NO:137:URELUMAB CDR3-L:
QQRSNWPPALT
SEQ ID NO:138:UTOMILUMAB CDR1-H:
YSFSTYWIS
SEQ ID NO:139:UTOMILUMAB CDR2-H:
KIYPGDSYTNYSPSFQG
SEQ ID NO:140:UTOMILUMAB CDR3-H:
GYGIFDY
SEQ ID NO:141:UTOMILUMAB CDR1-L:
SGDNIGDQYAH
SEQ ID NO:142:UTOMILUMAB CDR2-L:
QDKNRPS
SEQ ID NO:143:UTOMILUMAB CDR3-L:
ATYTGFGSLAV
SEQ ID NO:144:SEQ ID NO:121所示的蛋白质构建体的scFv:CD70(1F6)锚定结构域：
SEQ ID NO:145:SEQ ID NO:122所示的蛋白质构建体的scFv:CD70(2H5)锚定结构域：
SEQ ID NO:146:抗TRAILR2(Cona)-IgG1(N297A)-VH-scFv:CD70(9G2)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctgcaggaatctggccctggcctcgtgaagcctagccagaccctgagcctgacctgtaccgtgtctggcggcagcatcagcagcggcgactacttctggtcctggatcagacagctgcccggcaagggcctggaatggatcggccacatccacaacagcggcaccacctactacaaccccagcctgaagtccagagtgaccatcagcgtggacaccagcaagaagcagttcagcctgcggctgagcagcgtgacagccgccgatacagccgtgtactactgcgccagagacagaggcggcgattactactacggcatggacgtgtggggccagggcaccaccgtgaccgtgtctagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagccccggcaagctcgagcaggtgcagctggtggaatctggcggcggactgatgcagcctggcggctctctgagactgagctgtgccgccagcggcttcacctttagcagcagcgccatgagctgggtgcgccaggctcctggaaagggcctggaatgggtgtccagcatctacagcgacagcagctacacctactacgccgacagcgtgaagtcccggttcaccatcagcaccgacaacgccaagaacaccctgtacctgcagatgaacagcctgaagcccgacgacaccgccgtgtactactgtgccggcagcagcgattacgagggcagctttgcctcttggggccagggcacacaagtgaccgtgtcctccagatctagcaccaagggccccaagctggaagagggcgagttcagcgaggcccaattgcagagcgtcgtgacccagcctcctagcctgtctgcctctcctggaagcagcgtgcggctgacctgtacactgagcagcggcaacagcgtgggcaactacgacatcagctggtatcagcagaaggccggcagcccccccagatacctgctgtactactacagcgattccgtgaagcaccagggcagcggcgtgcccagcagattttccggaagctctgacgccagcgccaacgccggactgctgctgatttctggcctgcagcctgaggacgaggccgactactactgcagcgcctacaagagcggcagccacgtgttcggcggaggcaccaaactgacagtgctgggctaa
SEQ ID NO:157:SEQ ID NO:146所示的蛋白质构建体
SEQ ID NO:147:抗TRAILR2(Cona)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggagatcgtgctgacccagagccctggcaccctgtcactgtctccaggcgagagagccaccctgagctgtagagccagccagggcatcagccggtcttacctggcctggtatcagcagaagcccggccaggctcctagcctgctgatctacggcgccagcagcagagccaccggcatccccgatagattttccggcagcggctccggcaccgacttcaccctgacaatcagcagactggaacccgaggacttcgccgtgtattattgccagcagttcggcagcagcccctggacctttggccagggaacaaaagtgggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:158:SEQ ID NO:147所示的蛋白质构建体
SEQ ID NO:148:抗CD40(C)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctggttcagtctggcgccgaagtgaaaaagcctggcgcctctgtgaaggtgtcctgtacagccagcggcttcaacatcaaggactactacgtgcactgggtcaagcaggcccctggacaaggactggaatggatgggcagaatcgaccccgaggacggcgactctaagtacgcccctaagttccagggcaaagccaccatgaccgccgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcaccaccagctactatgtgggcacctacggctattggggccagggcacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:159:SEQ ID NO:148所示的蛋白质构建体
SEQ ID NO:149:抗CD40(C)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatccagatgacacagagccccagcagcctgtctgccagcgtgggagatagagtgaccatcacctgtagcgccagcagcagcgtgtcctacatgctgtggttccagcagaagcctggcaaggcccctaagctgctgatctacagcacctccaatctggccagcggcgtgccaagcagattttctggctctggcagcggcaccgacttcaccctgaccatatctagcctgcagccagaggacttcgccacctactactgccagcagcggacattctacccctacacctttggcggaggcaccaaggtggaaatcaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:160:SEQ ID NO:149所示的蛋白质构建体
SEQ ID NO:150:抗CD40(C)-Fab2-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtgcagctggttcagtctggcgccgaagtgaaaaagcctggcgcctctgtgaaggtgtcctgtacagccagcggcttcaacatcaaggactactacgtgcactgggtcaagcaggcccctggacaaggactggaatggatgggcagaatcgaccccgaggacggcgactctaagtacgcccctaagttccagggcaaagccaccatgaccgccgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcaccaccagctactatgtgggcacctacggctattggggccagggcacactggtcaccgtgtccagcagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:161:SEQ ID NO:150所示的蛋白质构建体
SEQ ID NO:151:抗41BB(HBBK)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:162:SEQ ID NO:151所示的蛋白质构建体
SEQ ID NO:152:抗41BB(HBBK)-Fab2-scFv:PD-L1(Ave)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatcctctagcgccagcacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgacagtgtcctggaactctggcgccctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgactgtgcccagcagcagcctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgccctcgaggaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcttctacaaagggccccaagctggaagagggcgagtttagcgaagcccaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgctttaa
SEQ ID NO:163:SEQ ID NO:152所示的蛋白质构建体
SEQ ID NO:153:抗PD-L1(Ave)-IgG2-VH-scFv:41BB(HBBK)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattcgaggtgcagctgctggaatctggcggaggacttgttcagcctggcggctctctgagactgtcttgtgccgccagcggcttcaccttcagcagctatatcatgatgtgggtccgacaggcccctggcaaaggccttgaatgggtgtccagcatctatcccagcggcggcatcaccttttacgccgacacagtgaagggcagattcaccatcagccgggacaacagcaagaacaccctgtacctgcagatgaacagcctgagagccgaggacaccgccgtgtactactgcgccagaatcaagctgggcaccgtgaccaccgtggattattggggacagggcaccctggtcaccgtgtcctccagatcctcgagtgctagcaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcggcgtgcacaccttcccagctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaagaattccaggtccagctgcagcagtctggcgccgaagttattaagcctggcgcctccgtgaagctgagctgtaaagccagcggctacaccttcagcagctactggatgcactgggtccgacaggctccaggacaaggcctggaatggatcggcgagatcaaccctggcaacggccacaccaactacaacgagaagttcaagagccgggccacactgaccggcgataccagcacaagcaccgtgtacatggaactgagcagcctgagaagcgaggacaccgccgtgtactactgcgccagatcctttaccaccgccagagcctttgcctattggggccagggaacactggtcaccgtgtccagcagatctagcacaaagggccccaagctggaagagggcgagtttagcgaggcccaattggacatcgtgatgactcagagccccgccttcctgtctgtgacccctggcgagaaagtgaccatcacctgtagagccagccagaccatcagcgactacctgcactggtatcagcagaagcccgatcaggcccctaagctgctgattaagtacgcctctcagagcatcagcggcatccccagcagattttctggcagcggctctggcaccgacttcacctttaccatcagctccctggaagccgaggatgccgccacctactattgtcaggacggccacagcttccctccaacctttggacagggcacaaagctggaaatcaagtaa
SEQ ID NO:164:SEQ ID NO:153所示的蛋白质构建体
SEQ ID NO:154:抗PD-L1(Ave)-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattgcagagcgccctgacacagcctgcatccgtgtctggatctccaggccagagcatcaccatctcttgtaccggcacaagctccgatgtcggcggctacaattacgtgtcctggtatcagcagcaccccggcaaggcccctaagctgatgatctacgacgtgtccaacagaccctccggcgtgtccaatagattcagcggcagcaagagcggcaacaccgccagcctgacaattagcggactgcaggccgaggacgaggccgattactactgtagcagctacaccagctcctccaccagagtgtttggcaccggcaccaaagtgaccgtgcttggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:165:SEQ ID NO:154所示的蛋白质构建体
SEQ ID NO:155:抗muCD27-IgG1(N297A)-VH-scFv:Fn14(18D1)
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattccaggtccagctgcagcagtctggcgccgaacttgtgaagcctggcagcagcgtgaagatcagctgtaaagccagcggctacaccttcaccagctacgacatgcactggatcaagcagcagcccggcaaaggcctggaatggatcggctggatctaccccggcaacggcaacaccaagtacaaccagaagttcaacggcaaggccacactgaccgccgacatctctagcagcacagcctacatgcagctgagcagcctgaccagcgaagatagcgccgtgtacttctgcgccaaatggggctacaacaacttcgactactggggccagggcgtgatggtcaccgtgtctagcagatccagcagcgcctctacaaagggccccagcgtgttccctctggcccctagcagcaagagcacatctggcggaacagccgccctgggctgcctcgtgaaggactactttcccgagcccgtgaccgtgtcctggaactctggcgctctgacaagcggcgtgcacacctttccagccgtgctgcagagcagcggcctgtactctctgagcagcgtcgtgacagtgcccagcagctctctgggcacccagacctacatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagaaggtggaacccaagagctgcgacaagacccacacctgtcccccttgtcctgcccccgaactgctgggaggcccttccgtgttcctgttccccccaaagcccaaggacaccctgatgatcagccggacccccgaagtgacctgcgtggtggtggatgtgtcccacgaggaccctgaagtgaagtttaattggtacgtggacggcgtggaagtgcacaacgccaagaccaagcctagagaggaacagtacgccagcacctaccgggtggtgtccgtgctgacagtgctgcaccaggactggctgaacggcaaagagtacaagtgcaaggtgtccaacaaggccctgcctgcccccatcgagaaaaccatcagcaaggccaagggccagccccgcgaaccccaggtgtacacactgcccccaagcagggacgagctgaccaagaaccaggtgtccctgacctgtctcgtgaaaggcttctaccccagcgatatcgccgtggaatgggagagcaacggccagcccgagaacaactacaagaccaccccccctgtgctggacagcgacggctcattcttcctgtacagcaagctgaccgtggacaagtcccggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaagctcgaggaggtgcagctggtggaatctggcggcggactggtgcagcctggcggatctctgagactgagctgtgccgccagcggcttcaccttcagcaactactggatgagctgggtgcgccaggcccctggcaaaggactggaatgggtgtccggcatcaacccaggcggcacctctacctactacgccgacagcgtgaagggccggttcaccatcagccgggacaacgccaagaacaccctgtacctgcagatgaacagcctgaagtccgaggacaccgccgtgtactactgcgccaagcacctgggcaactggggcgagtacaattactggggccagggcacacaagtgaccgtgtccagtagatctagcaccaagggccccaagctggaagagggcgagttcagcgaggcccaattgcagagcgccctgacccagcctccaagcgtgtcaggctctcctggcaagaccgtgaccatcagctgtgctggcaccggcggagatgtgggctacagaaacagcgtgtcctggtatcagcagctgcccggcatggcccccaaactgctgatctacgacgtggacaagcgggcctctggcatcaccgacagattcagcggcagcaagagcggcgataccgccagcctgacaatcagcggagtgcagagcgaggacgaggccgactactactgtgccagccagagaagcggaatcgccgccgtgtttggcggaggcacacacctgacagtgctgggctaa
SEQ ID NO:166:SEQ ID NO:155所示的蛋白质构建体
SEQ ID NO:156:抗muCD27-VL
atgaacttcggcttcagcctgatcttcctggtgctggtgctgaagggcgtgcagtgcgaagtgaagctggtgccccggcaattggactacaaggacgacgacgacaaagaattggacatccagatgacacagagccctgccagcctgtctgcctctctgggagagacagtgtccatcgattgtctggccagcgagggcatcagcaacgacctggcttggtatcagcagaagtccggcaagagccctcagctgctgatcaacagcgccagcagactggaagatggcgtgcccagcagattttctggctctggcagcggcacccggtacagcctgaagatttctggcatgcagcccgaggacgaggccgaatacttctgcctgcaaagctacagaagcccctggacctttggcggaggcacaaagctggaactgaagggatccgaaatcaagcgtacggtggccgctcccagcgtgttcatcttcccacctagcgacgagcagctgaagtccggcacagcctctgtcgtgtgcctgctgaacaacttctacccccgcgaggccaaggtgcagtggaaggtggacaatgccctgcagagcggcaacagccaggaaagcgtgaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcacaaggtgtacgcctgcgaagtgacccaccagggcctgtctagccccgtgaccaagagcttcaaccggggcgagtgctaa
SEQ ID NO:167:SEQ ID NO:156所示的蛋白质构建体
通过以下非限制性实施例进一步说明本发明。
实施例
实施例1：结合TNFR2的抗CD40 IgG1 G28.5的融合蛋白在结合TNFR2后表现出增强的CD40刺激活性。
分析IgG1抗体G28.5的融合蛋白(抗CD40(G28.5)-HC:scTNF80)，所述融合蛋白识别TNFRSF受体CD40，其单链结构域包含过短肽接头(scTNF80(mu)结构域)连接的三个拷贝的鼠源TNF的TNFR2特异性突变体，其中所述拷贝在分子内形成三聚体配体结构域，发明人意外观察到以下情况：
如果用抗CD40(G28.5)-HC:scTNF80刺激表达CD40的细胞或细胞共培养物，所述细胞或细胞共培养物未有FcγR或TNFR2表达，则在两种情况下都会导致没有或极小的CD40活化，例如反映在IL8的产生。相反，在表达FcγR的细胞的存在下，所述分子如预期的那样很好地活化了CD40(参见上述说明)。现在证明，与亲本G28.5抗体不同，抗CD40(G28.5)-HC:scTNF80在缺少FcγR的情况下，如果表达TNFR2，也可以以强烈的CD40活化方式起作用(图6)。显然，所述scTNF80结构域介导的抗CD40(G28.5)-HC:scTNF80与细胞结合的TNFR2的结合具有与Fc结构域介导的G28.5与表达FcγR的细胞的结合一样能产生激动性抗体的效应(图2)。这表明仅仅抗TNFRSF受体抗体或抗体融合蛋白的不依赖抗原结合结构域与细胞的结合足以分别赋予抗体和抗体融合蛋白高激动活性。
实施例2：抗TNFRSF受体CD40、CD95和TNFR2抗体与介导与细胞结合分子结合的蛋白结构域融合的融合蛋白，以不依赖FcγR的方式作为强激动剂。
根据仅仅抗TNFRSF受体抗体或抗体融合蛋白的不依赖抗原结合结构域的与细胞的结合足以分别赋予抗体和抗体融合蛋白高激动活性的想法，发明人还能证实识别TNFRSF受体CD40、CD95和TNFR2的抗体结构域与介导与细胞结合分子结合的蛋白结构域融合的其他抗体融合蛋白，能以不依赖FcγR的方式作为强激动剂。有关这些示例的详细信息，请参见图4、5、8和9，包括图例。
实施例3：抗TNFRSF受体CD40、CD95和TNFR2的抗体与靶向细胞表面抗原的scFv结构域的融合蛋白以不依赖FcγR的方式作为强激动剂。
另外，发明人研究了抗TNFRSF受体CD40、CD95和TNFR2的抗体与靶向细胞表面抗原的scFv结构域的融合蛋白，其分别靶向CD20(图10、11和12)和CD19(图7和12)。已发现这些融合蛋白以不依赖FcγR的方式起强激动剂的作用。有关这些示例的详细信息，请参见引用的图及其图例。
实施例4：抗TNFRSF受体CD40和CD95与scBaff的抗体融合蛋白以不依赖FcγR的方式作为强激动剂。
另外，发明人用scBaff研究了抗TNFRSF受体CD40(图13、14和15)和CD95(图16、17和18)的抗体或抗体片段的融合蛋白。已经发现，这些融合蛋白以不依赖FcγR的方式作为CD40和CD95的强激动剂，只要所述融合蛋白具有结合可被融合蛋白的scBaff结构域识别的表达细胞表面受体的可能性。这些示例的详细信息在上述图及其图例中给出。
实施例5：抗TNFRSF受体CD40、CD95、TNFR2和4-1BB的抗体的scFv融合蛋白以不依赖FcγR、scFv依赖的方式作为强激动剂。
此外，发明人研究了抗TNFRSF受体TNFR2(图19和20)、CD40(图21和22)、CD95(图21和22)和4-1BB(图21和22)的抗体的scFv融合蛋白。使用的scFv片段是scFv:CD20(图19、21和22)、scFv:CD70(2H5)(图20)和scFv:CD70(1F6)(图20)。发现这些融合蛋白作为被抗体部分(TNFR2、CD40、CD95、4-1BB)以不依赖FcγR的方式识别的TNFRSF受体的强激动剂，只要所述融合蛋白可以通过其scFv结构域结合被scFv结构域识别的细胞表面抗原(CD20、CD70)。这些示例的详细信息在上述图及其图例中给出。
实施例6：TNFRSF受体TRAILR2(DR5)的scFv融合蛋白以不依赖FcγR、scFv依赖的方式作为强激动剂。
此外，发明人研究了抗TNFRSF受体TRAILR2的抗体(Conatumumab)的scFv融合蛋白，也称为DR5(图23)。所使用的scFv片段是scFv:CD70。发现所述融合蛋白被抗体部分(TRAILR2)以不依赖FcγR的方式识别的TNFRSF受体的强激动剂，只要所述融合蛋白具有通过其scFv结构域与可被scFv结构域识别的细胞表面抗原CD70结合的可能性。
实施例7：抗抗原呈递细胞刺激的TNFRSF受体CD40或T细胞刺激的TNFRSF受体4-1BB的抗体的融合蛋白，其具有来自阻断PD-1L和PD-1相互作用的检查点抑制剂Avelumab的scFv作为锚定结构域，以不依赖FcγR、scFv:PD-1L依赖的方式起强激动剂作用。发明人还研究了抗TNFRSF受体CD40(图24)和4-1BB(图25)的抗体或抗体片段与scFv:PD-1L的融合蛋白，所述scFv:PD-1L是检查点抑制剂Avelumab的scFv，其中Avelumab为一种阻断PD-1L与PD-1结合的抗体。发现这些融合蛋白可以以不依赖FcγR的方式作为CD40和4-1BB的强激动剂，只要所述融合蛋白可能与被融合蛋白的scFv:PD-1L结构域识别的细胞表面表达的PD-1L结合。因此，这些融合蛋白是结合了免疫检查点阻断与免疫检查点阻断依赖性限制性激活免疫刺激因子TNFRSF受体的双功能抗体示例。这些示例的详细信息在上述图及其图例中给出。
实施例8：当与锚定于细胞表面抗原的完整抗体融合时，对TNFRSF受体Fn14和4-1BB特异的scFv以不依赖FcγR的方式作为强激动剂。另外，发明人用识别PD-1L或鼠CD27的抗体研究了抗TNFRSF受体4-1BB(图26)和Fn14(图27)的scFv融合蛋白。已经发现，只要所述融合蛋白的抗体部分有可能结合其细胞表面表达的抗原PD-1L和鼠CD27时，所述融合蛋白以不依赖FcγR的方式作为4-1BB和Fn14的强激动剂。这些示例展示了完整的抗体可以用作锚定结构域，以使抗TNFRSF受体抗体片段(在此为scFv)能作为具有锚定依赖性活性的激动剂。这些示例的详细信息在上述图及其图例中给出。
实施例结论
对于用于不依赖FcγR和抗原结合域的细胞相互作用，并且与抗体融合的结构域，在下文中也称为固定结构域，没有特别限制。因此，本发明人能生产利用IL2配体或scTNFSF配体进行细胞结合的TRAAFFIAA，但也可以产生出于所述目的使用异源scFv抗体片段的TRAAFFIAA。为了获得激动活性，所述抗原结合结构域以不依赖FcγR和抗原结合域的方式结合细胞似乎也是足够的。例如，已经通过与CD20特异性scFv融合进行基因工程改造的CD95特异性抗体E09的Fab2片段(图11)，显示出依赖CD20的强烈增加的TNFRSF受体刺激活性。
从本发明人的工作中，立即可显而易见的是，使用在TRAAFFIAA中使用的经适当选择的固定化结构域，可以在体内获得限于靶结构的局部激动效应。因此，有可能避免可能限制治疗的全身副作用。因此，TRAAFFIAA不仅可以有利地以不依赖FcγR的方式用抗体刺激TNFRSF受体，而且还开拓了包括临床应用的新的应用领域。
工业适用性
根据本发明，所述抗体融合蛋白及其组合物可以工业化生产并作为商业产品出售，例如作为药物。因此，本发明在工业上是可应用的。
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SEQUENCE LISTING
<110> 尤利乌斯·马克西米利安维尔茨堡大学
<120> 具有非FcγR依赖性激动活性的肿瘤坏死因子（TNF）受体超家族（TNFRSF）
受体-激活抗体融合蛋白（具有非FcγR依赖性激动活性的TNFRSF受体-激活
抗体融合蛋白；TRAAFFIAA）
<130> P20113345WP
<150> EP 17210849.0
<151> 2017-12-28
<150> EP 18189937.8
<151> 2018-08-21
<160> 167
<170> PatentIn version 3.5
<210> 1
<211> 2892
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scTNF80(mu)-pCR3 (N297A)
<400> 1
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca aggaattcga gttcacacgg gacaaacctg tggctcatgt ggtggccaat 1500
catcaggtgg aggaacagct ggaatggctg agtcagagag caaacgccct gctggcaaat 1560
gggatggacc tcaaagacaa tcagctcgtg gtgcctgccg atggactgta cctggtgtac 1620
tctcaggtcc tgtttaaggg acagggatgc cccgattacg tgctgctcac ccacactgtg 1680
tcacgcttcg ccatctcata ccaggagaaa gtcaatctcc tctccgccgt gaaatcacca 1740
tgtcctaagg atactcccga gggagccgaa ctgaaacctt ggtacgaacc catctacctg 1800
ggcggcgtgt ttcagctgga gaaaggcgat cagctctccg ccgaagtgaa tctgcccaaa 1860
tacctcaact ttagggaatc cggacaggtc tactttggcg tgattgccct gggaggcgga 1920
tctggaggag gctctggcgg gggatctggg ggcggatccg acaaacctgt ggcacacgtc 1980
gtggcaaacc atcaggtcga ggaacagctc gagtggctgt cacagagggc caatgccctg 2040
ctggcaaatg gaatggatct gaaggataat cagctcgtcg tgcctgccga cggcctctac 2100
ctcgtctact ctcaggtcct ctttaaggga cagggctgcc ccgactacgt cctgctcact 2160
cataccgtga gtcgcttcgc tatttcatac caggaaaaag tcaacctgct gagtgctgtg 2220
aaatctcctt gccctaagga tacccctgag ggagccgaac tcaaaccatg gtacgagcca 2280
atctacctcg gaggagtgtt tcagctggaa aaaggggatc agctctccgc cgaagtcaac 2340
ctccccaaat acctcaattt ccgggaatcc ggacaggtgt actttggagt cattgccctg 2400
ggaggcggct ctggcggggg atctggagga ggctccggag gaggcagtga caaacccgtc 2460
gctcacgtgg tggcaaatca tcaggtcgag gaacagctgg aatggctgtc tcagagagca 2520
aacgctctcc tcgccaatgg aatggatctc aaggacaacc agctcgtcgt ccctgccgat 2580
ggactctacc tggtctactc tcaggtgctc tttaagggac agggatgccc cgattacgtc 2640
ctgctcacac acaccgtgtc tcgctttgct atttcatacc aggagaaagt caatctgctg 2700
tctgccgtca aatctccttg tccaaaagac acacccgagg gagccgaact caaaccttgg 2760
tacgagccaa tctacctggg gggagtgttt cagctggaga agggggatca gctctccgcc 2820
gaagtgaatc tcccaaaata cctcaatttt cgggaatccg gacaggtcta ctttggagtg 2880
attgccctgt ag 2892
<210> 2
<211> 1455
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-pCR3 (hIgG1)
<400> 2
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca agtaa 1455
<210> 3
<211> 2189
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scFv-抗CD20-pCR3 (N297A)
<400> 3
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca aggaattcca ggtacaactg cagcagcctg gggctgagct ggtgaagcct 1500
ggggcctcag tgaagatgtc ctgcaaggct tctggctaca catttaccag ttacaatatg 1560
cactgggtaa aacagacacc tggtcggggc ctggaatgga ttggagctat ttatcccgga 1620
aatggtgata cttcctacaa tcagaagttc aaaggcaagg ccacattgac tgcagacaaa 1680
tcctccagca cagcctacat gcagctcagc agcctgacat ctgaggactc tgcggtctat 1740
tactgtgcaa gatcgactta ctacggcggt gactggtact tcaatgtctg gggcgcaggg 1800
accacggtca ccgtctcttc aggaggaggc ggatccggcg gaggcggaag cggtggcgga 1860
ggctctcaaa ttgttctctc ccagtctcca gcaatcctgt ctgcatctcc aggggagaag 1920
gtcacaatga cttgcagggc cagctcaagt gtaagttaca tccactggtt ccagcagaag 1980
ccaggatcct cccccaaacc ctggatttat gccacatcca acctggcttc tggagtccct 2040
gttcgcttca gtggcagtgg gtctgggact tcttactctc tcacaatcag cagagtggag 2100
gcgaagatgc tgccacttat tactgccagc agtggactag taacccaccc acgttcggag 2160
gggggaccaa gctggaaatc aaacgttaa 2189
<210> 4
<211> 2874
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VH-heavy-full-scBaff-pCR3 (N297A)
<400> 4
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatccagca gcgcctctac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 840
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 900
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 960
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1020
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1080
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1140
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1200
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1260
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1320
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1380
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1440
agccccggca agctcgaggg acccgaggaa actgtgactc aggactgtct ccagctcatt 1500
gccgatagtg aaacccctac catccagaaa ggctcttaca ccttcgtgcc atggctgctg 1560
tcattcaaac ggggatctgc tctggaggag aaggaaaaca aaatcctggt caaggaaacc 1620
ggctacttct tcatctacgg ccaggtcctc tacaccgaca aaacatacgc tatggggcat 1680
ctcattcagc ggaaaaaagt ccacgtgttc ggcgacgaac tctctctcgt gacactgttc 1740
cggtgtattc agaacatgcc cgagactctg cccaataata gctgctactc tgctggcatt 1800
gccaaactgg aggagggcga cgaactccag ctggctattc ctagggaaaa tgcccagatt 1860
agcctggacg gggatgtgac attttttggc gccctgaaac tgctgggagg cggagggagt 1920
ggcgggggag gctctggacc tgaggaaact gtgacccagg attgtctcca gctcattgcc 1980
gatagtgaga ctcctaccat tcagaaggga tcttacacct ttgtgccttg gctgctgtct 2040
ttcaaacggg gctctgctct ggaggaaaag gagaacaaaa ttctggtcaa agagactggc 2100
tacttcttca tctacggcca ggtgctgtac accgacaaaa catacgccat gggccatctc 2160
attcagcgga aaaaagtcca cgtgttcggc gacgaactct ctctcgtgac actgttccgg 2220
tgtatccaga acatgcccga gacactgccc aataatagct gctactctgc cggcattgct 2280
aaactggagg agggggacga actccagctg gctattccta gggaaaatgc ccagatttct 2340
ctcgatgggg atgtgacatt cttcggggcc ctcaaactgc tgggaggcgg cggatctggc 2400
ggaggcggga gtcaattcgc agcaggtcca gaagaaacag tcactcaaga ctgcttgcaa 2460
ctgattgcag acagtgaaac accaactata caaaaaggat cttacacatt tgttccatgg 2520
cttctcagct ttaaaagggg aagtgcccta gaagaaaaag agaataaaat attggtcaaa 2580
gaaactggtt acttttttat atatggtcag gttttatata ctgataagac ctacgccatg 2640
ggacatctaa ttcagaggaa gaaggtccat gtctttgggg atgaattgag tctggtgact 2700
ttgtttcgat gtattcaaaa tatgcctgaa acactaccca ataattcctg ctattcagct 2760
ggcattgcaa aactggaaga aggagatgaa ctccaacttg caataccaag agaaaatgca 2820
caaatatcac tggatggaga tgtcacattt tttggtgcat tgaaactgct gtga 2874
<210> 5
<211> 789
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40-Flag-VL-light-full-pCR3
<400> 5
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
gtgatgactc agaacccact gtctctgcct gtgtctctgg gggatgaggc tagcatttct 180
tgccgctcat ctcagtcact ggagaactcc aatggcaaca ccttcctgaa ttggtttttc 240
cagaaacccg gccagtcacc tcagctgctc atctaccgag tgagcaatcg gtttagcgga 300
gtgcccgatc gattctctgg ctccggatct gggaccgact ttaccctgaa aatctcacga 360
gtggaggccg aggatctggg agtgtacttc tgtctccagg tcacacatgt gccttacaca 420
tttggcggcg gaacaactct cgaaatcaaa ggatccgaaa tcaagcgtac ggtggccgct 480
cccagcgtgt tcatcttccc acctagcgac gagcagctga agtccggcac agcctctgtc 540
gtgtgcctgc tgaacaactt ctacccccgc gaggccaagg tgcagtggaa ggtggacaat 600
gccctgcaga gcggcaacag ccaggaaagc gtgaccgagc aggacagcaa ggactccacc 660
tacagcctga gcagcaccct gaccctgagc aaggccgact acgagaagca caaggtgtac 720
gcctgcgaag tgacccacca gggcctgtct agccccgtga ccaagagctt caaccggggc 780
gagtgctaa 789
<210> 6
<211> 1908
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-full-IL2(mu)-pCR3 (hIgG1) (无 Flag)
<400> 6
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctct agcgccagca caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgacagtg tcctggaact ctggcgccct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
actgtgccca gcagcagcct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccctga actgctgggc ggaccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcccagaga ggaacagtac aacagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgcctcc cagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta cccctccgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattgc ccggtaccgc ccctaccagc agcagcacct ctagctctac agccgaggct 1500
caacaacaac aacaacagca acagcagcag cagcagcacc tggaacagct gctgatggac 1560
ctgcaggaac tgctgagccg gatggaaaac taccggaacc tgaagctgcc ccggatgctg 1620
accttcaagt tctacctgcc caagcaggcc accgagctga aggatctgca gtgcctggaa 1680
gatgagctgg gccccctgag acacgtgctg gatctgaccc agagcaagag ctttcagctg 1740
gaagatgccg agaacttcat cagcaacatc agagtgaccg tcgtgaagct gaagggcagc 1800
gacaacacct tcgagtgcca gttcgacgac gagagcgcta ccgtggtgga cttcctgcgg 1860
agatggatcg ccttctgcca gagcatcatc agcaccagcc cccagtaa 1908
<210> 7
<211> 2199
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-konst-full-scFv-抗CD19-pCR3 (N297A) (无Flag)
<400> 7
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg acattcagat gacgcagtct ccatcctcca tgtctgtatc tctgggagac 1500
acagtcagca tcacttgcca tgcaagtcag ggcattagca gtaatatagg gtggttgcag 1560
cagaaaccag ggaaatcatt taagggcctg atctatcatg gaaccaactt ggaagatgga 1620
gttccatcaa ggttcagtgg cagtggatct ggagcagatt attctctcac catcagcagc 1680
ctggaatctg aagattttgc agactattac tgtgtacagt atgctcagtt tccgtacacg 1740
ttcggagggg ggaccaagct ggagctgaaa cgtggtggtg gtggttctgg tggtggtggt 1800
tctggcggcg gcggctccag tggtggtgga tcccaggttc agctgcagca atctggacct 1860
gagctggtga agcctggggc ctcagtgaag atttcctgca aagcttctgg ctacgcattc 1920
agtagctctt ggatggactg ggtgaagcag aggcctggac agggtcttga gtggattgga 1980
cggatttatc ctggagatgg agatactaac tacaatggga agttcaaggg caaggccaca 2040
ctgactgcag acaaatcctc cagcacagcc tacatgcagc tcagcagcct gacctctgtg 2100
gactctgcgg tctatttctg tgcaaggtcc attactacgg tagtagggtg gtacttcgat 2160
gtctggggcg cagggaccac ggtcaccgtt tcctcctaa 2199
<210> 8
<211> 2181
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-konst-full-scFv-抗CD20-pCR3 (N297A) (无Flag)
<400> 8
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc aggtacaact gcagcagcct ggggctgagc tggtgaagcc tggggcctca 1500
gtgaagatgt cctgcaaggc ttctggctac acatttacca gttacaatat gcactgggta 1560
aaacagacac ctggtcgggg cctggaatgg attggagcta tttatcccgg aaatggtgat 1620
acttcctaca atcagaagtt caaaggcaag gccacattga ctgcagacaa atcctccagc 1680
acagcctaca tgcagctcag cagcctgaca tctgaggact ctgcggtcta ttactgtgca 1740
agatcgactt actacggcgg tgactggtac ttcaatgtct ggggcgcagg gaccacggtc 1800
accgtctctt caggaggagg cggatccggc ggaggcggaa gcggtggcgg aggctctcaa 1860
attgttctct cccagtctcc agcaatcctg tctgcatctc caggggagaa ggtcacaatg 1920
acttgcaggg ccagctcaag tgtaagttac atccactggt tccagcagaa gccaggaccc 1980
tcccccaaac cctggattta tgccacatcc aacctggctt ctggagtccc tgttcgcttc 2040
agtggcagtg ggtctgggac ttcttactct ctcacaatca gcagagtgga ggctgaagat 2100
gctgccactt attactgcca gcagtggact agtaacccac ccacgttcgg aggggggacc 2160
aagctggaaa tcaaacgtta a 2181
<210> 9
<211> 1446
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-pCR3 (IgG1) (无Flag)
<400> 9
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctct agcgccagca caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgacagtg tcctggaact ctggcgccct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
actgtgccca gcagcagcct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccctga actgctgggc ggaccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcccagaga ggaacagtac aacagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgcctcc cagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta cccctccgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aagtaa 1446
<210> 10
<211> 2724
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-scGITRL-pCR3 (N297A)
<400> 10
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg gcggagggag cggacagctg gaaactgcca aagaaccctg tatggccaaa 1500
ttcggaccac tgcctagcaa atggcagatg gctagtagcg aacctccatg tgtgaacaaa 1560
gtgagcgatt ggaaactcga gatcctccag aatggactgt acctcatcta cggacaggtc 1620
gccccaaatg ccaattacaa tgatgtggcc ccctttgaag tccggctcta caaaaacaag 1680
gatatgatcc agaccctcac caacaaatcc aaaatccaga atgtgggcgg cacatacgaa 1740
ctccatgtcg gcgataccat cgatctcatt ttcaactctg aacaccaggt gctcaaaaac 1800
aacacctact ggggaatcat cctgctggca aaccctcagt tcatctccgg cggcggctct 1860
ggcggcggat ctggcggagg gagtggcgga ggctcacagc tggagactgc taaagaaccc 1920
tgtatggcaa aattcgggcc cctgccctca aaatggcaga tggcctcctc tgaaccaccc 1980
tgtgtgaaca aagtgagtga ttggaaactg gaaatcctcc agaacggcct ctacctcatc 2040
tacggacagg tggcacccaa tgccaattac aacgacgtgg cacccttcga agtgagactg 2100
tacaaaaaca aggatatgat ccagaccctc accaacaaat caaaaatcca gaatgtcgga 2160
gggacatacg aactccatgt gggagacact atcgacctca ttttcaattc cgaacatcag 2220
gtcctgaaaa acaacactta ctggggcatt attctgctcg ccaatccaca gtttattagt 2280
ggaggcgggg gatctggggg aggctccggc ggagggagtg gaggcggatc tcaattccaa 2340
ttagagactg ctaaggagcc ctgtatggct aagtttggac cattaccctc aaaatggcaa 2400
atggcatctt ctgaacctcc ttgcgtgaat aaggtgtctg actggaagct ggagatactt 2460
cagaatggct tatatttaat ttatggccaa gtggctccca atgcaaacta caatgatgta 2520
gctccttttg aggtgcggct gtataaaaac aaagacatga tacaaactct aacaaacaaa 2580
tctaaaatcc aaaatgtagg agggacttat gaattgcatg ttggggacac catagacttg 2640
atattcaact ctgagcatca ggttctaaaa aataatacat actggggtat cattttacta 2700
gcaaatcccc aattcatctc ctag 2724
<210> 11
<211> 3411
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-sc(mu)41BBL-pCR3 (N297A)
<400> 11
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcg gcggtggaag tggtactgaa ccgcgacccg ctctcactat aaccaccagc 1500
cctaacttgg gaacgcggga gaataacgcg gatcaagtaa ccccagtttc ccatatcggg 1560
tgtccaaata ccactcaaca aggatcaccg gtctttgcta agctcctggc gaaaaaccaa 1620
gcgtccctct gtaatacaac ccttaattgg cactcccaag atggggcggg ttcttcatac 1680
ctctcacagg ggctccgata cgaagaagac aaaaaggagt tggtggtcga ctcaccagga 1740
ctttactatg tattcctgga gttgaagctc tctcctacat tcaccaatac cggtcacaaa 1800
gtacagggtt gggtgagcct tgtgctccag gcgaagccgc aggtggacga cttcgacaac 1860
cttgcactca ctgtagagct gttcccttgt tccatggaga ataaattggt ggaccgctct 1920
tggagtcagc tcctcctcct caaggcgggt catcgattga gcgttggcct tcgggcatat 1980
cttcatgggg cgcaagatgc ataccgcgat tgggaattgt cataccccaa cacgacctct 2040
tttggcctgt ttctggtcaa acccgacaat ccgtgggaag gtgggggaag cggagggggt 2100
tcaggaggag gatctggggg aggttcaact gaaccgaggc ccgcgcttac tatcacgact 2160
tccccaaatc tggggactag ggaaaacaat gccgatcagg tcactcctgt tagtcacatt 2220
ggttgtccta atacgaccca gcaaggctct ccggtgtttg ccaaactgtt ggccaaaaat 2280
caggcgtcac tttgtaatac aacgctcaac tggcatagtc aggatggggc cggctcctca 2340
tacttgtctc aaggtcttag gtacgaagaa gataagaagg agctggtggt agacagcccc 2400
gggctctact atgtgttcct ggagctcaaa ctgtcaccga cgttcactaa caccggtcat 2460
aaggtacagg gttgggtatc cttggtgttg caagcaaaac cccaggtgga cgatttcgat 2520
aatcttgcgc ttactgtaga gctctttcca tgttcaatgg aaaataaact ggtcgatagg 2580
agctggtccc aacttctcct tcttaaagct ggccatcgcc tgagtgttgg cctgagagcg 2640
tatcttcatg gggcgcagga cgcttaccgg gattgggaac tgtcataccc aaacaccacc 2700
agctttgggc tcttccttgt aaagccagac aatccgtggg aggggggagg cgggagtggg 2760
ggcgggtctg gagggggcag tggggggggt agtacggagc cgcgccccgc cctgaccatc 2820
acaacgtcac ccaatcttgg gactcgggag aataacgccg accaggttac ccctgtatcc 2880
catatcggtt gtcctaatac gacacaacaa ggcagtcctg tattcgctaa actcttggca 2940
aaaaaccagg ccagtctttg taatacgacg cttaattggc atagccagga cggtgcgggc 3000
agctcctacc tttcccaggg gctcaggtat gaagaagata agaaagaact cgttgtagac 3060
agtcccggat tgtattacgt ttttttggaa ctcaagctct ctccaacctt caccaatacg 3120
ggacataagg tccagggctg ggtgagcctc gtactccagg ctaagccgca agttgacgat 3180
ttcgataatc tcgctcttac agtggagttg tttccctgta gtatggagaa taagctcgtc 3240
gaccggtctt ggagccaact tctgctgctt aaggctggtc accggctcag tgtaggcctc 3300
cgagcgtatt tgcatggggc gcaggacgcc tatcgagact gggagctttc ctaccctaac 3360
acgaccagct ttggactctt cttggtgaaa cctgacaatc cgtgggaata a 3411
<210> 12
<211> 1836
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-HC-heavy-full-(mu)GITRL-pCR3 (N297A)
<400> 12
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc caactgccat cgagtcctgc atggttaagt ttgaactatc atcctcaaaa 1500
tggcacatga catctcccaa acctcactgt gtgaatacga catctgatgg gaagctgaag 1560
atactgcaga gtggcacata tttaatctac ggccaagtga ttcctgtgga taagaaatac 1620
ataaaagaca atgccccctt cgtagtacag atatataaaa agaatgatgt cctacaaact 1680
ctaatgaatg attttcaaat cttgcctata ggaggggttt atgaactgca tgctggagat 1740
aacatatatc tgaagttcaa ctctaaagac catattcaga aaactaacac atactggggg 1800
atcatcttaa tgcctgatct accattcatc tcttag 1836
<210> 13
<211> 735
<212> DNA
<213> Artificial Sequence
<220>
<223> C4-LC-light-full-pCR3 (无Flag)
<400> 13
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggacatt gtgatgaccc agtctcacaa attcatgtcc 120
acatcagtag gagacagggt cagcatcacc tgcaaggcca gtcaggatgt ggatactgct 180
gtagcctggt atcaacaaaa accagggcaa tctcctaaac tactgattta ctgggcatcc 240
acccggcaca ctggagtccc tgatcgcttc acaggcagtg gatctgggac agattatact 300
ctcaccatca gcagtgtgca ggctgaagac ctggcgcgtt attactgtca gcaatattat 360
agtgttcctc cgacgttcgg tggaggcacc aagctgggat ccgaaatcaa gcgtacggtg 420
gccgctccca gcgtgttcat cttcccacct agcgacgagc agctgaagtc cggcacagcc 480
tctgtcgtgt gcctgctgaa caacttctac ccccgcgagg ccaaggtgca gtggaaggtg 540
gacaatgccc tgcagagcgg caacagccag gaaagcgtga ccgagcagga cagcaaggac 600
tccacctaca gcctgagcag caccctgacc ctgagcaagg ccgactacga gaagcacaag 660
gtgtacgcct gcgaagtgac ccaccagggc ctgtctagcc ccgtgaccaa gagcttcaac 720
cggggcgagt gctaa 735
<210> 14
<211> 2268
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-Flag-VH-full-scFv-抗CD19-pCR3 (IgG1)
<400> 14
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc agatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agctcgagga cattcagatg acgcagtctc catcctccat gtctgtatct 1560
ctgggagaca cagtcagcat cacttgccat gcaagtcagg gcattagcag taatataggg 1620
tggttgcagc agaaaccagg gaaatcattt aagggcctga tctatcatgg aaccaacttg 1680
gaagatggag ttccatcaag gttcagtggc agtggatctg gagcagatta ttctctcacc 1740
atcagcagcc tggaatctga agattttgca gactattact gtgtacagta tgctcagttt 1800
ccgtacacgt tcggaggggg gaccaagctg gagctgaaac gtggtggtgg tggttctggt 1860
ggtggtggtt ctggcggcgg cggctccagt ggtggtggat cccaggttca gctgcagcaa 1920
tctggacctg agctggtgaa gcctggggcc tcagtgaaga tttcctgcaa agcttctggc 1980
tacgcattca gtagctcttg gatggactgg gtgaagcaga ggcctggaca gggtcttgag 2040
tggattggac ggatttatcc tggagatgga gatactaact acaatgggaa gttcaagggc 2100
aaggccacac tgactgcaga caaatcctcc agcacagcct acatgcagct cagcagcctg 2160
acctctgtgg actctgcggt ctatttctgt gcaaggtcca ttactacggt agtagggtgg 2220
tacttcgatg tctggggcgc agggaccacg gtcaccgttt cctcctaa 2268
<210> 15
<211> 1515
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VH-heavy-full-pCR3 (IgG1)
<400> 15
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc agatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccctgaa ctgctgggcg gaccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcccagagag gaacagtaca acagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgcctccc agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac ccctccgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agtaa 1515
<210> 16
<211> 1602
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-Flag-VH-FAB2(1-114)-scFv-抗CD20-pCR3 (IgG1)
<400> 16
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgccctcgag caggtacaac tgcagcagcc tggggctgag 900
ctggtgaagc ctggggcctc agtgaagatg tcctgcaagg cttctggcta cacatttacc 960
agttacaata tgcactgggt aaaacagaca cctggtcggg gcctggaatg gattggagct 1020
atttatcccg gaaatggtga tacttcctac aatcagaagt tcaaaggcaa ggccacattg 1080
actgcagaca aatcctccag cacagcctac atgcagctca gcagcctgac atctgaggac 1140
tctgcggtct attactgtgc aagatcgact tactacggcg gtgactggta cttcaatgtc 1200
tggggcgcag ggaccacggt caccgtctct tcaggaggag gcggatccgg cggaggcgga 1260
agcggtggcg gaggctctca aattgttctc tcccagtctc cagcaatcct gtctgcatct 1320
ccaggggaga aggtcacaat gacttgcagg gccagctcaa gtgtaagtta catccactgg 1380
ttccagcaga agccaggatc ctcccccaaa ccctggattt atgccacatc caacctggct 1440
tctggagtcc ctgttcgctt cagtggcagt gggtctggga cttcttactc tctcacaatc 1500
agcagagtgg aggctgaaga tgctgccact tattactgcc agcagtggac tagtaaccca 1560
cccacgttcg gaggggggac caagctggaa atcaaacgtt aa 1602
<210> 17
<211> 2934
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VH-heavy-full-scBaff-pCR3 (N297A)
<400> 17
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatccagca gcgcctctac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca agctcgaggg acccgaggaa actgtgactc aggactgtct ccagctcatt 1560
gccgatagtg aaacccctac catccagaaa ggctcttaca ccttcgtgcc atggctgctg 1620
tcattcaaac ggggatctgc tctggaggag aaggaaaaca aaatcctggt caaggaaacc 1680
ggctacttct tcatctacgg ccaggtcctc tacaccgaca aaacatacgc tatggggcat 1740
ctcattcagc ggaaaaaagt ccacgtgttc ggcgacgaac tctctctcgt gacactgttc 1800
cggtgtattc agaacatgcc cgagactctg cccaataata gctgctactc tgctggcatt 1860
gccaaactgg aggagggcga cgaactccag ctggctattc ctagggaaaa tgcccagatt 1920
agcctggacg gggatgtgac attttttggc gccctgaaac tgctgggagg cggagggagt 1980
ggcgggggag gctctggacc tgaggaaact gtgacccagg attgtctcca gctcattgcc 2040
gatagtgaga ctcctaccat tcagaaggga tcttacacct ttgtgccttg gctgctgtct 2100
ttcaaacggg gctctgctct ggaggaaaag gagaacaaaa ttctggtcaa agagactggc 2160
tacttcttca tctacggcca ggtgctgtac accgacaaaa catacgccat gggccatctc 2220
attcagcgga aaaaagtcca cgtgttcggc gacgaactct ctctcgtgac actgttccgg 2280
tgtatccaga acatgcccga gacactgccc aataatagct gctactctgc cggcattgct 2340
aaactggagg agggggacga actccagctg gctattccta gggaaaatgc ccagatttct 2400
ctcgatgggg atgtgacatt cttcggggcc ctcaaactgc tgggaggcgg cggatctggc 2460
ggaggcggga gtcaattcgc agcaggtcca gaagaaacag tcactcaaga ctgcttgcaa 2520
ctgattgcag acagtgaaac accaactata caaaaaggat cttacacatt tgttccatgg 2580
cttctcagct ttaaaagggg aagtgcccta gaagaaaaag agaataaaat attggtcaaa 2640
gaaactggtt acttttttat atatggtcag gttttatata ctgataagac ctacgccatg 2700
ggacatctaa ttcagaggaa gaaggtccat gtctttgggg atgaattgag tctggtgact 2760
ttgtttcgat gtattcaaaa tatgcctgaa acactaccca ataattcctg ctattcagct 2820
ggcattgcaa aactggaaga aggagatgaa ctccaacttg caataccaag agaaaatgca 2880
caaatatcac tggatggaga tgtcacattt tttggtgcat tgaaactgct gtga 2934
<210> 18
<211> 786
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95-E09-Flag-VL-light-full-pCR3
<400> 18
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagagc 120
gtgctgaccc agcctcctag cgtgtccgaa gcccctagac agaccgtgac catcagctgc 180
tccggcaaca gcttcaacat cggcagatac cccgtgaact ggtatcagca gctgccaggc 240
aaggccccta aactgctgat ctactataac aacctgcggt tcagcggagt gtccgaccgg 300
ttctctggca gcaagtctgg cacatctgcc agcctggcca tccgggatct gctgtctgag 360
gacgaggccg actactactg cagcacctgg gacgacaccc tgaagggctg ggtgttcggc 420
ggaggcacca aagtgacagt gctgggcgga tccgaaatca agcgtacggt ggccgctccc 480
agcgtgttca tcttcccacc tagcgacgag cagctgaagt ccggcacagc ctctgtcgtg 540
tgcctgctga acaacttcta cccccgcgag gccaaggtgc agtggaaggt ggacaatgcc 600
ctgcagagcg gcaacagcca ggaaagcgtg accgagcagg acagcaagga ctccacctac 660
agcctgagca gcaccctgac cctgagcaag gccgactacg agaagcacaa ggtgtacgcc 720
tgcgaagtga cccaccaggg cctgtctagc cccgtgacca agagcttcaa ccggggcgag 780
tgctaa 786
<210> 19
<211> 963
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 1表达的蛋白质构建体
<400> 19
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Glu Phe Glu Phe Thr Arg Asp Lys Pro Val Ala His
485 490 495
Val Val Ala Asn His Gln Val Glu Glu Gln Leu Glu Trp Leu Ser Gln
500 505 510
Arg Ala Asn Ala Leu Leu Ala Asn Gly Met Asp Leu Lys Asp Asn Gln
515 520 525
Leu Val Val Pro Ala Asp Gly Leu Tyr Leu Val Tyr Ser Gln Val Leu
530 535 540
Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val Leu Leu Thr His Thr Val
545 550 555 560
Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys Val Asn Leu Leu Ser Ala
565 570 575
Val Lys Ser Pro Cys Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys
580 585 590
Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys
595 600 605
Gly Asp Gln Leu Ser Ala Glu Val Asn Leu Pro Lys Tyr Leu Asn Phe
610 615 620
Arg Glu Ser Gly Gln Val Tyr Phe Gly Val Ile Ala Leu Gly Gly Gly
625 630 635 640
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Asp Lys Pro
645 650 655
Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln Leu Glu Trp
660 665 670
Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met Asp Leu Lys
675 680 685
Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu Val Tyr Ser
690 695 700
Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val Leu Leu Thr
705 710 715 720
His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys Val Asn Leu
725 730 735
Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro Glu Gly Ala
740 745 750
Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly Val Phe Gln
755 760 765
Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu Pro Lys Tyr
770 775 780
Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val Ile Ala Leu
785 790 795 800
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
805 810 815
Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln
820 825 830
Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met
835 840 845
Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu
850 855 860
Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val
865 870 875 880
Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys
885 890 895
Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro
900 905 910
Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly
915 920 925
Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu
930 935 940
Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val
945 950 955 960
Ile Ala Leu
<210> 20
<211> 484
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 2表达的蛋白质构建体
<400> 20
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys
<210> 21
<211> 729
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 3表达的蛋白质构建体
<400> 21
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln Pro Gly Ala Glu
485 490 495
Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly
500 505 510
Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly
515 520 525
Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr
530 535 540
Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys
545 550 555 560
Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp
565 570 575
Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp
580 585 590
Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly
595 600 605
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile
610 615 620
Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys
625 630 635 640
Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp
645 650 655
Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr
660 665 670
Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser
675 680 685
Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala
690 695 700
Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly
705 710 715 720
Gly Gly Thr Lys Leu Glu Ile Lys Arg
725
<210> 22
<211> 957
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 4表达的蛋白质构建体
<400> 22
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
260 265 270
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
275 280 285
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
290 295 300
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
305 310 315 320
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr
325 330 335
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
340 345 350
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
355 360 365
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
370 375 380
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
385 390 395 400
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
405 410 415
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
420 425 430
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
435 440 445
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
450 455 460
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
465 470 475 480
Ser Pro Gly Lys Leu Glu Gly Pro Glu Glu Thr Val Thr Gln Asp Cys
485 490 495
Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser
500 505 510
Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu
515 520 525
Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe
530 535 540
Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His
545 550 555 560
Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu
565 570 575
Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn
580 585 590
Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu
595 600 605
Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly
610 615 620
Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser
625 630 635 640
Gly Gly Gly Gly Ser Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu
645 650 655
Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr
660 665 670
Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu
675 680 685
Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile
690 695 700
Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu
705 710 715 720
Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val
725 730 735
Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn
740 745 750
Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu
755 760 765
Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp
770 775 780
Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly
785 790 795 800
Gly Gly Gly Ser Gln Phe Ala Ala Gly Pro Glu Glu Thr Val Thr Gln
805 810 815
Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys
820 825 830
Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser
835 840 845
Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr
850 855 860
Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met
865 870 875 880
Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu
885 890 895
Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu
900 905 910
Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly
915 920 925
Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu
930 935 940
Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu
945 950 955
<210> 23
<211> 262
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 5表达的蛋白质构建体
<400> 23
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Val Met Thr Gln Asn Pro Leu Ser
35 40 45
Leu Pro Val Ser Leu Gly Asp Glu Ala Ser Ile Ser Cys Arg Ser Ser
50 55 60
Gln Ser Leu Glu Asn Ser Asn Gly Asn Thr Phe Leu Asn Trp Phe Phe
65 70 75 80
Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Val Ser Asn
85 90 95
Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr
100 105 110
Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val
115 120 125
Tyr Phe Cys Leu Gln Val Thr His Val Pro Tyr Thr Phe Gly Gly Gly
130 135 140
Thr Thr Leu Glu Ile Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala
145 150 155 160
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
165 170 175
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
180 185 190
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
195 200 205
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
210 215 220
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
225 230 235 240
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
245 250 255
Phe Asn Arg Gly Glu Cys
260
<210> 24
<211> 635
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 6表达的蛋白质构建体
<400> 24
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Leu Pro Gly Thr Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser
485 490 495
Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln
500 505 510
His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser Arg Met
515 520 525
Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe Lys Phe
530 535 540
Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu
545 550 555 560
Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln Ser Lys
565 570 575
Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile Arg Val
580 585 590
Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe
595 600 605
Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp Ile Ala
610 615 620
Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln
625 630 635
<210> 25
<211> 732
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 7表达的蛋白质构建体
<400> 25
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val
485 490 495
Ser Leu Gly Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile
500 505 510
Ser Ser Asn Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys
515 520 525
Gly Leu Ile Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg
530 535 540
Phe Ser Gly Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser
545 550 555 560
Leu Glu Ser Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln
565 570 575
Phe Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly
580 585 590
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly
595 600 605
Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys
610 615 620
Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe
625 630 635 640
Ser Ser Ser Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu
645 650 655
Glu Trp Ile Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn
660 665 670
Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
675 680 685
Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val
690 695 700
Tyr Phe Cys Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp
705 710 715 720
Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser
725 730
<210> 26
<211> 726
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 8表达的蛋白质构建体
<400> 26
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys
485 490 495
Pro Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe
500 505 510
Thr Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu
515 520 525
Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn
530 535 540
Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser
545 550 555 560
Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val
565 570 575
Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn
580 585 590
Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly
595 600 605
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser
610 615 620
Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met
625 630 635 640
Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln
645 650 655
Lys Pro Gly Pro Ser Ser Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu
660 665 670
Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser
675 680 685
Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr
690 695 700
Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr
705 710 715 720
Lys Leu Glu Ile Lys Arg
725
<210> 27
<211> 481
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 9表达的蛋白质构建体
<400> 27
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys
<210> 28
<211> 907
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 10表达的蛋白质构建体
<400> 28
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gly Gly Gly Ser Gly Gln Leu Glu Thr Ala Lys Glu Pro
485 490 495
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
500 505 510
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
515 520 525
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
530 535 540
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
545 550 555 560
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
565 570 575
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
580 585 590
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
595 600 605
Leu Ala Asn Pro Gln Phe Ile Ser Gly Gly Gly Ser Gly Gly Gly Ser
610 615 620
Gly Gly Gly Ser Gly Gly Gly Ser Gln Leu Glu Thr Ala Lys Glu Pro
625 630 635 640
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
645 650 655
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
660 665 670
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
675 680 685
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
690 695 700
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
705 710 715 720
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
725 730 735
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
740 745 750
Leu Ala Asn Pro Gln Phe Ile Ser Gly Gly Gly Gly Ser Gly Gly Gly
755 760 765
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gln Phe Gln Leu Glu Thr Ala
770 775 780
Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln
785 790 795 800
Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys
805 810 815
Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala
820 825 830
Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr
835 840 845
Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln
850 855 860
Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu
865 870 875 880
Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly
885 890 895
Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
900 905
<210> 29
<211> 1136
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 11表达的蛋白质构建体
<400> 29
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gly Gly Gly Ser Gly Thr Glu Pro Arg Pro Ala Leu Thr
485 490 495
Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln
500 505 510
Val Thr Pro Val Ser His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly
515 520 525
Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys
530 535 540
Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr
545 550 555 560
Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val
565 570 575
Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro
580 585 590
Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val
595 600 605
Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr
610 615 620
Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser
625 630 635 640
Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val Gly
645 650 655
Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp Trp Glu
660 665 670
Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu Val Lys Pro
675 680 685
Asp Asn Pro Trp Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
690 695 700
Ser Gly Gly Gly Ser Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr
705 710 715 720
Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro
725 730 735
Val Ser His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val
740 745 750
Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr
755 760 765
Leu Asn Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln
770 775 780
Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro
785 790 795 800
Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr
805 810 815
Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val Leu Gln Ala
820 825 830
Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr Val Glu Leu
835 840 845
Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser Trp Ser Gln
850 855 860
Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val Gly Leu Arg Ala
865 870 875 880
Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp Trp Glu Leu Ser Tyr
885 890 895
Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu Val Lys Pro Asp Asn Pro
900 905 910
Trp Glu Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly
915 920 925
Gly Gly Ser Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro
930 935 940
Asn Leu Gly Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro Val Ser
945 950 955 960
His Ile Gly Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val Phe Ala
965 970 975
Lys Leu Leu Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn
980 985 990
Trp His Ser Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu
995 1000 1005
Arg Tyr Glu Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro Gly
1010 1015 1020
Leu Tyr Tyr Val Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr
1025 1030 1035
Asn Thr Gly His Lys Val Gln Gly Trp Val Ser Leu Val Leu Gln
1040 1045 1050
Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu Ala Leu Thr Val
1055 1060 1065
Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val Asp Arg Ser
1070 1075 1080
Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu Ser Val
1085 1090 1095
Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg Asp
1100 1105 1110
Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu
1115 1120 1125
Val Lys Pro Asp Asn Pro Trp Glu
1130 1135
<210> 30
<211> 611
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 12表达的蛋白质构建体
<400> 30
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Pro Thr Ala Ile Glu Ser Cys Met Val Lys Phe Glu Leu
485 490 495
Ser Ser Ser Lys Trp His Met Thr Ser Pro Lys Pro His Cys Val Asn
500 505 510
Thr Thr Ser Asp Gly Lys Leu Lys Ile Leu Gln Ser Gly Thr Tyr Leu
515 520 525
Ile Tyr Gly Gln Val Ile Pro Val Asp Lys Lys Tyr Ile Lys Asp Asn
530 535 540
Ala Pro Phe Val Val Gln Ile Tyr Lys Lys Asn Asp Val Leu Gln Thr
545 550 555 560
Leu Met Asn Asp Phe Gln Ile Leu Pro Ile Gly Gly Val Tyr Glu Leu
565 570 575
His Ala Gly Asp Asn Ile Tyr Leu Lys Phe Asn Ser Lys Asp His Ile
580 585 590
Gln Lys Thr Asn Thr Tyr Trp Gly Ile Ile Leu Met Pro Asp Leu Pro
595 600 605
Phe Ile Ser
610
<210> 31
<211> 244
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 13表达的蛋白质构建体
<400> 31
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Ile Val Met
20 25 30
Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser
35 40 45
Ile Thr Cys Lys Ala Ser Gln Asp Val Asp Thr Ala Val Ala Trp Tyr
50 55 60
Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser
65 70 75 80
Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly
85 90 95
Thr Asp Tyr Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
100 105 110
Arg Tyr Tyr Cys Gln Gln Tyr Tyr Ser Val Pro Pro Thr Phe Gly Gly
115 120 125
Gly Thr Lys Leu Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
130 135 140
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
145 150 155 160
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
165 170 175
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
180 185 190
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
195 200 205
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
210 215 220
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
225 230 235 240
Arg Gly Glu Cys
<210> 32
<211> 755
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 14表达的蛋白质构建体
<400> 32
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Asp Ile Gln Met Thr Gln
500 505 510
Ser Pro Ser Ser Met Ser Val Ser Leu Gly Asp Thr Val Ser Ile Thr
515 520 525
Cys His Ala Ser Gln Gly Ile Ser Ser Asn Ile Gly Trp Leu Gln Gln
530 535 540
Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile Tyr His Gly Thr Asn Leu
545 550 555 560
Glu Asp Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp
565 570 575
Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser Glu Asp Phe Ala Asp Tyr
580 585 590
Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr Thr Phe Gly Gly Gly Thr
595 600 605
Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
610 615 620
Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser Gln Val Gln Leu Gln Gln
625 630 635 640
Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys
645 650 655
Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser Trp Met Asp Trp Val Lys
660 665 670
Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Arg Ile Tyr Pro Gly
675 680 685
Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu
690 695 700
Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu
705 710 715 720
Thr Ser Val Asp Ser Ala Val Tyr Phe Cys Ala Arg Ser Ile Thr Thr
725 730 735
Val Val Gly Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr
740 745 750
Val Ser Ser
755
<210> 33
<211> 504
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 15表达的蛋白质构建体
<400> 33
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys
500
<210> 34
<211> 533
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 16表达的蛋白质构建体
<400> 34
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Leu Glu Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro
290 295 300
Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
305 310 315 320
Ser Tyr Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu
325 330 335
Trp Ile Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln
340 345 350
Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr
355 360 365
Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr
370 375 380
Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val
385 390 395 400
Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser
405 410 415
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln
420 425 430
Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr
435 440 445
Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys
450 455 460
Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala
465 470 475 480
Ser Gly Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr
485 490 495
Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr
500 505 510
Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys
515 520 525
Leu Glu Ile Lys Arg
530
<210> 35
<211> 977
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 17表达的蛋白质构建体
<400> 35
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Gly Pro Glu Glu Thr Val
500 505 510
Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile
515 520 525
Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg
530 535 540
Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr
545 550 555 560
Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr
565 570 575
Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp
580 585 590
Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu
595 600 605
Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu
610 615 620
Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile
625 630 635 640
Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Pro Glu Glu Thr Val Thr
660 665 670
Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln
675 680 685
Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly
690 695 700
Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly
705 710 715 720
Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala
725 730 735
Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe Gly Asp Glu
740 745 750
Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr
755 760 765
Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu
770 775 780
Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser
785 790 795 800
Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu Leu Gly Gly
805 810 815
Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe Ala Ala Gly Pro Glu Glu
820 825 830
Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr Pro
835 840 845
Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser Phe
850 855 860
Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val Lys
865 870 875 880
Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp Lys
885 890 895
Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val Phe
900 905 910
Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn Met
915 920 925
Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala Lys
930 935 940
Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn Ala
945 950 955 960
Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys Leu
965 970 975
Leu
<210> 36
<211> 261
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 18表达的蛋白质构建体
<400> 36
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Ser Val Leu Thr Gln Pro Pro Ser Val
35 40 45
Ser Glu Ala Pro Arg Gln Thr Val Thr Ile Ser Cys Ser Gly Asn Ser
50 55 60
Phe Asn Ile Gly Arg Tyr Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly
65 70 75 80
Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Asn Asn Leu Arg Phe Ser Gly
85 90 95
Val Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu
100 105 110
Ala Ile Arg Asp Leu Leu Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser
115 120 125
Thr Trp Asp Asp Thr Leu Lys Gly Trp Val Phe Gly Gly Gly Thr Lys
130 135 140
Val Thr Val Leu Gly Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro
145 150 155 160
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
165 170 175
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
180 185 190
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
195 200 205
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
210 215 220
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
225 230 235 240
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
245 250 255
Asn Arg Gly Glu Cys
260
<210> 37
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR1-H
<400> 37
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn
1 5 10
<210> 38
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR2-H
<400> 38
Tyr Ile Arg Tyr Asp Gly Thr Ser Glu Tyr Thr Pro Ser Leu Lys Asn
1 5 10 15
<210> 39
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR1-L
<400> 39
Ser Ser Gln Ser Leu Glu Asn Ser Asn Gly Asn Thr Phe Leu Asn
1 5 10 15
<210> 40
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5抗体的CDR2-L
<400> 40
Arg Val Ser Asn Arg Phe Ser
1 5
<210> 41
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD40 G28.5的CDR3-L
<400> 41
Leu Gln Val Thr His Val Pro Tyr Thr
1 5
<210> 42
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR1-H
<400> 42
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val
1 5 10
<210> 43
<211> 21
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR2-H
<400> 43
Ser Ile Ala Tyr Arg Gly Asn Ser Asn Ser Gly Ser Thr Tyr Tyr Asn
1 5 10 15
Pro Ser Leu Lys Ser
20
<210> 44
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR3-H
<400> 44
Arg Gln Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp
1 5 10 15
Val
<210> 45
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR1-L
<400> 45
Ser Gly Asn Ser Phe Asn Ile Gly Arg Tyr Pro Val Asn
1 5 10
<210> 46
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR2-L
<400> 46
Tyr Asn Asn Leu Arg Phe Ser
1 5
<210> 47
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD95 E09抗体的CDR3-L
<400> 47
Ser Thr Trp Asp Asp Thr Leu Lys Gly Trp Val
1 5 10
<210> 48
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR1-H
<400> 48
Gly Ser Ile Ser Ser Gly Asp Tyr Phe Trp Ser
1 5 10
<210> 49
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR2-H
<400> 49
His Ile His Asn Ser Gly Thr Thr Tyr Tyr Asn Pro Ser Leu Lys Ser
1 5 10 15
<210> 50
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR3-H
<400> 50
Asp Arg Gly Gly Asp Tyr Tyr Tyr Gly Met Asp Val
1 5 10
<210> 51
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR1-L
<400> 51
Arg Ala Ser Gln Gly Ile Ser Arg Ser Tyr Leu Ala
1 5 10
<210> 52
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR2-L
<400> 52
Gly Ala Ser Ser Arg Ala Thr
1 5
<210> 53
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> DR5 Conatu抗体的CDR3-L
<400> 53
Gln Gln Phe Gly Ser Ser Pro Trp Thr
1 5
<210> 54
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR1-H
<400> 54
Tyr Thr Phe Thr Asp Tyr Gly Met His
1 5
<210> 55
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR2-H
<400> 55
Val Ile Ser Thr Tyr Asn Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 56
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR3-H
<400> 56
Ala Tyr Tyr Gly Asn Leu Tyr Tyr Ala Met Asp Tyr
1 5 10
<210> 57
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR1-L
<400> 57
Arg Ala Ser Lys Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His
1 5 10 15
<210> 58
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR2-L
<400> 58
Tyr Ala Ser Asn Leu Glu Ser
1 5
<210> 59
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 P4A8抗体的CDR3-L
<400> 59
Gln His Ser Arg Glu Leu Pro Phe Thr
1 5
<210> 60
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR1-H
<400> 60
Tyr Ser Phe Thr Ser Tyr Asp Ile Asn
1 5
<210> 61
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR2-H
<400> 61
Trp Ile Tyr Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys
1 5 10 15
Gly
<210> 62
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR3-H
<400> 62
Leu Thr Gly Pro Tyr Trp Tyr Phe Asp Val
1 5 10
<210> 63
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR1-L
<400> 63
Lys Ala Ser Gln Asp Val Asp Thr Ala Val Ala
1 5 10
<210> 64
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR2-L
<400> 64
Trp Ala Ser Thr Arg His Thr
1 5
<210> 65
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> TNFR2 C4抗体的CDR3-L
<400> 65
Gln Gln Tyr Tyr Ser Val Pro Pro Thr
1 5
<210> 66
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR1-H
<400> 66
Tyr Thr Phe Thr Ser Tyr Asn Met His
1 5
<210> 67
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR2-H
<400> 67
Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 68
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR3-H
<400> 68
Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val
1 5 10
<210> 69
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR1-L
<400> 69
Arg Ala Ser Ser Ser Val Ser Tyr Ile His
1 5 10
<210> 70
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR2-L
<400> 70
Ala Thr Ser Asn Leu Ala Ser
1 5
<210> 71
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD20抗体的CDR3-L
<400> 71
Gln Gln Trp Thr Ser Asn Pro Pro Thr
1 5
<210> 72
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR1-H
<400> 72
Tyr Ala Phe Ser Ser Ser Trp Met Asp
1 5
<210> 73
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR2-H
<400> 73
Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys
1 5 10 15
Gly
<210> 74
<211> 12
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR3-H
<400> 74
Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val
1 5 10
<210> 75
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR1-L
<400> 75
His Ala Ser Gln Gly Ile Ser Ser Asn Ile Gly
1 5 10
<210> 76
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR2-L
<400> 76
His Gly Thr Asn Leu Glu Asp
1 5
<210> 77
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD19抗体的CDR3-L
<400> 77
Val Gln Tyr Ala Gln Phe Pro Tyr Thr
1 5
<210> 78
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR1-H
<400> 78
Tyr Thr Phe Thr Asn Tyr Gly Met Asn
1 5
<210> 79
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR2-H
<400> 79
Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe Lys
1 5 10 15
Gly
<210> 80
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR3-H
<400> 80
Asp Tyr Gly Asp Tyr Gly Met Asp Tyr
1 5
<210> 81
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR1-L
<400> 81
Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His
1 5 10 15
<210> 82
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR2-L
<400> 82
Leu Ala Ser Asn Leu Glu Ser
1 5
<210> 83
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 1F6抗体的CDR3-L
<400> 83
Gln His Ser Arg Glu Val Pro Trp Thr
1 5
<210> 84
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR1-H
<400> 84
Phe Thr Phe Ser Ser Tyr Trp Met Ser
1 5
<210> 85
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR2-H
<400> 85
Glu Ile Arg Leu Lys Ser Asp Asn Tyr Ala Thr His Tyr Ala Glu Ser
1 5 10 15
Val Lys Gly
<210> 86
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR3-H
<400> 86
Gly Tyr Tyr Ala Asp Ala Met Asp Tyr
1 5
<210> 87
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR1-L
<400> 87
Arg Ala Ser Gln Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His
1 5 10 15
<210> 88
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR2-L
<400> 88
Tyr Ala Ser Asn Leu Glu Ser
1 5
<210> 89
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Fn14 PDL192抗体的CDR3-L
<400> 89
Gln His Ser Trp Glu Ile Pro Tyr Thr
1 5
<210> 90
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR1-H
<400> 90
Phe Thr Phe Ser Ser Tyr Ile Met His
1 5
<210> 91
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR2-H
<400> 91
Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 92
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR3-H
<400> 92
Asp Thr Asp Gly Tyr Asp Phe Asp Tyr
1 5
<210> 93
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR1-L
<400> 93
Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
1 5 10
<210> 94
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR2-L
<400> 94
Asp Ala Ser Asn Arg Ala Thr
1 5
<210> 95
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> CD70 2H5抗体的CDR3-L
<400> 95
Gln Gln Arg Thr Asn Trp Pro Leu Thr
1 5
<210> 96
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 21所示蛋白质构建体的scFv-抗CD20锚定结构域
<400> 96
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 97
<211> 149
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 24所示的蛋白质构建体的鼠源 IL-2 锚定结构域
<400> 97
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln
145
<210> 98
<211> 249
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 25所示的蛋白质构建体的scFv-抗CD19锚定结构域
<400> 98
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val Ser Leu Gly
1 5 10 15
Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile Ser Ser Asn
20 25 30
Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile
35 40 45
Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser
65 70 75 80
Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser
115 120 125
Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
130 135 140
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser
145 150 155 160
Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
165 170 175
Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe
180 185 190
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
195 200 205
Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val Tyr Phe Cys
210 215 220
Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val Trp Gly
225 230 235 240
Ala Gly Thr Thr Val Thr Val Ser Ser
245
<210> 99
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 26所示的蛋白质构建体的scFv-抗CD20锚定结构域
<400> 99
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Pro Ser Ser Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 100
<211> 249
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 32所示的蛋白质构建体的scFv-抗CD19锚定结构域
<400> 100
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Val Ser Leu Gly
1 5 10 15
Asp Thr Val Ser Ile Thr Cys His Ala Ser Gln Gly Ile Ser Ser Asn
20 25 30
Ile Gly Trp Leu Gln Gln Lys Pro Gly Lys Ser Phe Lys Gly Leu Ile
35 40 45
Tyr His Gly Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Ala Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser
65 70 75 80
Glu Asp Phe Ala Asp Tyr Tyr Cys Val Gln Tyr Ala Gln Phe Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Gly Ser
115 120 125
Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
130 135 140
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser
145 150 155 160
Trp Met Asp Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
165 170 175
Gly Arg Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe
180 185 190
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
195 200 205
Met Gln Leu Ser Ser Leu Thr Ser Val Asp Ser Ala Val Tyr Phe Cys
210 215 220
Ala Arg Ser Ile Thr Thr Val Val Gly Trp Tyr Phe Asp Val Trp Gly
225 230 235 240
Ala Gly Thr Thr Val Thr Val Ser Ser
245
<210> 101
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 34所示的蛋白质构建体的scFv-抗CD20锚定结构域
<400> 101
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys Arg
<210> 102
<211> 473
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 19所示的蛋白质构建体的scTNF80（鼠）锚定结构域
<400> 102
Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val Glu Glu Gln
1 5 10 15
Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala Asn Gly Met
20 25 30
Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly Leu Tyr Leu
35 40 45
Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro Asp Tyr Val
50 55 60
Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr Gln Glu Lys
65 70 75 80
Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys Asp Thr Pro
85 90 95
Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly
100 105 110
Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu Val Asn Leu
115 120 125
Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr Phe Gly Val
130 135 140
Ile Ala Leu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly
145 150 155 160
Gly Gly Ser Asp Lys Pro Val Ala His Val Val Ala Asn His Gln Val
165 170 175
Glu Glu Gln Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala Leu Leu Ala
180 185 190
Asn Gly Met Asp Leu Lys Asp Asn Gln Leu Val Val Pro Ala Asp Gly
195 200 205
Leu Tyr Leu Val Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly Cys Pro
210 215 220
Asp Tyr Val Leu Leu Thr His Thr Val Ser Arg Phe Ala Ile Ser Tyr
225 230 235 240
Gln Glu Lys Val Asn Leu Leu Ser Ala Val Lys Ser Pro Cys Pro Lys
245 250 255
Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu Pro Ile Tyr
260 265 270
Leu Gly Gly Val Phe Gln Leu Glu Lys Gly Asp Gln Leu Ser Ala Glu
275 280 285
Val Asn Leu Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly Gln Val Tyr
290 295 300
Phe Gly Val Ile Ala Leu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Ser Gly Gly Gly Ser Asp Lys Pro Val Ala His Val Val Ala Asn
325 330 335
His Gln Val Glu Glu Gln Leu Glu Trp Leu Ser Gln Arg Ala Asn Ala
340 345 350
Leu Leu Ala Asn Gly Met Asp Leu Lys Asp Asn Gln Leu Val Val Pro
355 360 365
Ala Asp Gly Leu Tyr Leu Val Tyr Ser Gln Val Leu Phe Lys Gly Gln
370 375 380
Gly Cys Pro Asp Tyr Val Leu Leu Thr His Thr Val Ser Arg Phe Ala
385 390 395 400
Ile Ser Tyr Gln Glu Lys Val Asn Leu Leu Ser Ala Val Lys Ser Pro
405 410 415
Cys Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr Glu
420 425 430
Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys Gly Asp Gln Leu
435 440 445
Ser Ala Glu Val Asn Leu Pro Lys Tyr Leu Asn Phe Arg Glu Ser Gly
450 455 460
Gln Val Tyr Phe Gly Val Ile Ala Leu
465 470
<210> 103
<211> 471
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 22所示的蛋白质构建体的scBaff锚定结构域
<400> 103
Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp
1 5 10 15
Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp
20 25 30
Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys
35 40 45
Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu
50 55 60
Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys
65 70 75 80
Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys
85 90 95
Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala
100 105 110
Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro
115 120 125
Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly
130 135 140
Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
145 150 155 160
Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser
165 170 175
Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu
180 185 190
Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile
195 200 205
Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr
210 215 220
Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val
225 230 235 240
His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile
245 250 255
Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly
260 265 270
Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg
275 280 285
Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala
290 295 300
Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe
305 310 315 320
Ala Ala Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile
325 330 335
Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val
340 345 350
Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu
355 360 365
Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln
370 375 380
Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg
385 390 395 400
Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe
405 410 415
Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr
420 425 430
Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala
435 440 445
Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe
450 455 460
Phe Gly Ala Leu Lys Leu Leu
465 470
<210> 104
<211> 419
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 28所示的蛋白质构建体的scGITRL锚定结构域
<400> 104
Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu
1 5 10 15
Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys
20 25 30
Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile
35 40 45
Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe
50 55 60
Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn
65 70 75 80
Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly
85 90 95
Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn
100 105 110
Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
115 120 125
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
130 135 140
Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe Gly Pro Leu
145 150 155 160
Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys
165 170 175
Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile
180 185 190
Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe
195 200 205
Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn
210 215 220
Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly
225 230 235 240
Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn
245 250 255
Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
260 265 270
Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly
275 280 285
Ser Gln Phe Gln Leu Glu Thr Ala Lys Glu Pro Cys Met Ala Lys Phe
290 295 300
Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys
305 310 315 320
Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu
325 330 335
Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val
340 345 350
Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr
355 360 365
Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu
370 375 380
His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val
385 390 395 400
Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln
405 410 415
Phe Ile Ser
<210> 105
<211> 648
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 29所示的蛋白质构建体的鼠sc41BBL锚定结构域
<400> 105
Thr Glu Pro Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly
1 5 10 15
Thr Arg Glu Asn Asn Ala Asp Gln Val Thr Pro Val Ser His Ile Gly
20 25 30
Cys Pro Asn Thr Thr Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu
35 40 45
Ala Lys Asn Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser
50 55 60
Gln Asp Gly Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu
65 70 75 80
Glu Asp Lys Lys Glu Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val
85 90 95
Phe Leu Glu Leu Lys Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys
100 105 110
Val Gln Gly Trp Val Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp
115 120 125
Asp Phe Asp Asn Leu Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met
130 135 140
Glu Asn Lys Leu Val Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys
145 150 155 160
Ala Gly His Arg Leu Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala
165 170 175
Gln Asp Ala Tyr Arg Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser
180 185 190
Phe Gly Leu Phe Leu Val Lys Pro Asp Asn Pro Trp Glu Gly Gly Gly
195 200 205
Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Glu Pro
210 215 220
Arg Pro Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu
225 230 235 240
Asn Asn Ala Asp Gln Val Thr Pro Val Ser His Ile Gly Cys Pro Asn
245 250 255
Thr Thr Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn
260 265 270
Gln Ala Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly
275 280 285
Ala Gly Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys
290 295 300
Lys Glu Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu
305 310 315 320
Leu Lys Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly
325 330 335
Trp Val Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp
340 345 350
Asn Leu Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys
355 360 365
Leu Val Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His
370 375 380
Arg Leu Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala
385 390 395 400
Tyr Arg Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu
405 410 415
Phe Leu Val Lys Pro Asp Asn Pro Trp Glu Gly Gly Gly Gly Ser Gly
420 425 430
Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Thr Glu Pro Arg Pro
435 440 445
Ala Leu Thr Ile Thr Thr Ser Pro Asn Leu Gly Thr Arg Glu Asn Asn
450 455 460
Ala Asp Gln Val Thr Pro Val Ser His Ile Gly Cys Pro Asn Thr Thr
465 470 475 480
Gln Gln Gly Ser Pro Val Phe Ala Lys Leu Leu Ala Lys Asn Gln Ala
485 490 495
Ser Leu Cys Asn Thr Thr Leu Asn Trp His Ser Gln Asp Gly Ala Gly
500 505 510
Ser Ser Tyr Leu Ser Gln Gly Leu Arg Tyr Glu Glu Asp Lys Lys Glu
515 520 525
Leu Val Val Asp Ser Pro Gly Leu Tyr Tyr Val Phe Leu Glu Leu Lys
530 535 540
Leu Ser Pro Thr Phe Thr Asn Thr Gly His Lys Val Gln Gly Trp Val
545 550 555 560
Ser Leu Val Leu Gln Ala Lys Pro Gln Val Asp Asp Phe Asp Asn Leu
565 570 575
Ala Leu Thr Val Glu Leu Phe Pro Cys Ser Met Glu Asn Lys Leu Val
580 585 590
Asp Arg Ser Trp Ser Gln Leu Leu Leu Leu Lys Ala Gly His Arg Leu
595 600 605
Ser Val Gly Leu Arg Ala Tyr Leu His Gly Ala Gln Asp Ala Tyr Arg
610 615 620
Asp Trp Glu Leu Ser Tyr Pro Asn Thr Thr Ser Phe Gly Leu Phe Leu
625 630 635 640
Val Lys Pro Asp Asn Pro Trp Glu
645
<210> 106
<211> 128
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 30所示的蛋白质构建体的鼠GITRL锚定结构域
<400> 106
Pro Thr Ala Ile Glu Ser Cys Met Val Lys Phe Glu Leu Ser Ser Ser
1 5 10 15
Lys Trp His Met Thr Ser Pro Lys Pro His Cys Val Asn Thr Thr Ser
20 25 30
Asp Gly Lys Leu Lys Ile Leu Gln Ser Gly Thr Tyr Leu Ile Tyr Gly
35 40 45
Gln Val Ile Pro Val Asp Lys Lys Tyr Ile Lys Asp Asn Ala Pro Phe
50 55 60
Val Val Gln Ile Tyr Lys Lys Asn Asp Val Leu Gln Thr Leu Met Asn
65 70 75 80
Asp Phe Gln Ile Leu Pro Ile Gly Gly Val Tyr Glu Leu His Ala Gly
85 90 95
Asp Asn Ile Tyr Leu Lys Phe Asn Ser Lys Asp His Ile Gln Lys Thr
100 105 110
Asn Thr Tyr Trp Gly Ile Ile Leu Met Pro Asp Leu Pro Phe Ile Ser
115 120 125
<210> 107
<211> 111
<212> PRT
<213> Artificial Sequence
<220>
<223> 人IL-2锚定结构域
<400> 107
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Glu His Lys Pro Ser Ser Gln Arg Lys Glu Glu Ser Thr Cys
100 105 110
<210> 108
<211> 807
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(G28.5)-VH(1-114)
<400> 108
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgcctaa 807
<210> 109
<211> 2226
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(G28.5)-VH(1-114)-scBaff
<400> 109
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggatatc 120
cagctccagc agtctggccc tggactcgtc aaaccatctc agagcctgtc tctcacctgt 180
tctgtcaccg gatactccat caccaccaac tacaactgga attggattcg gcagtttcct 240
gggaacaaac tcgaatggat gggatacatc cgatacgacg gcactagtga atacacccca 300
tctctcaaaa atcgggtgtc cattacccgg gacacttcta tgaaccagtt ctttctccga 360
ctcacctctg tgacacctga ggataccgcc acatactact gtgctagact ggactactgg 420
gggcagggaa cactggtgac cgtgtcatct ggatcctcta gcgccagcac aaagggcccc 480
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 540
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 600
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 660
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 720
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 780
cacacctgtc ccccttgtcc tgccctcgag ggacccgagg aaactgtgac tcaggactgt 840
ctccagctca ttgccgatag tgaaacccct accatccaga aaggctctta caccttcgtg 900
ccatggctgc tgtcattcaa acggggatct gctctggagg agaaggaaaa caaaatcctg 960
gtcaaggaaa ccggctactt cttcatctac ggccaggtcc tctacaccga caaaacatac 1020
gctatggggc atctcattca gcggaaaaaa gtccacgtgt tcggcgacga actctctctc 1080
gtgacactgt tccggtgtat tcagaacatg cccgagactc tgcccaataa tagctgctac 1140
tctgctggca ttgccaaact ggaggagggc gacgaactcc agctggctat tcctagggaa 1200
aatgcccaga ttagcctgga cggggatgtg acattttttg gcgccctgaa actgctggga 1260
ggcggaggga gtggcggggg aggctctgga cctgaggaaa ctgtgaccca ggattgtctc 1320
cagctcattg ccgatagtga gactcctacc attcagaagg gatcttacac ctttgtgcct 1380
tggctgctgt ctttcaaacg gggctctgct ctggaggaaa aggagaacaa aattctggtc 1440
aaagagactg gctacttctt catctacggc caggtgctgt acaccgacaa aacatacgcc 1500
atgggccatc tcattcagcg gaaaaaagtc cacgtgttcg gcgacgaact ctctctcgtg 1560
acactgttcc ggtgtatcca gaacatgccc gagacactgc ccaataatag ctgctactct 1620
gccggcattg ctaaactgga ggagggggac gaactccagc tggctattcc tagggaaaat 1680
gcccagattt ctctcgatgg ggatgtgaca ttcttcgggg ccctcaaact gctgggaggc 1740
ggcggatctg gcggaggcgg gagtcaattc gcagcaggtc cagaagaaac agtcactcaa 1800
gactgcttgc aactgattgc agacagtgaa acaccaacta tacaaaaagg atcttacaca 1860
tttgttccat ggcttctcag ctttaaaagg ggaagtgccc tagaagaaaa agagaataaa 1920
atattggtca aagaaactgg ttactttttt atatatggtc aggttttata tactgataag 1980
acctacgcca tgggacatct aattcagagg aagaaggtcc atgtctttgg ggatgaattg 2040
agtctggtga ctttgtttcg atgtattcaa aatatgcctg aaacactacc caataattcc 2100
tgctattcag ctggcattgc aaaactggaa gaaggagatg aactccaact tgcaatacca 2160
agagaaaatg cacaaatatc actggatgga gatgtcacat tttttggtgc attgaaactg 2220
ctgtga 2226
<210> 110
<211> 2286
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-VH(1-114)-scBaff
<400> 110
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatcctcta gcgccagcac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgacagtgt cctggaactc tggcgccctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga ctgtgcccag cagcagcctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgccctcgag ggacccgagg aaactgtgac tcaggactgt 900
ctccagctca ttgccgatag tgaaacccct accatccaga aaggctctta caccttcgtg 960
ccatggctgc tgtcattcaa acggggatct gctctggagg agaaggaaaa caaaatcctg 1020
gtcaaggaaa ccggctactt cttcatctac ggccaggtcc tctacaccga caaaacatac 1080
gctatggggc atctcattca gcggaaaaaa gtccacgtgt tcggcgacga actctctctc 1140
gtgacactgt tccggtgtat tcagaacatg cccgagactc tgcccaataa tagctgctac 1200
tctgctggca ttgccaaact ggaggagggc gacgaactcc agctggctat tcctagggaa 1260
aatgcccaga ttagcctgga cggggatgtg acattttttg gcgccctgaa actgctggga 1320
ggcggaggga gtggcggggg aggctctgga cctgaggaaa ctgtgaccca ggattgtctc 1380
cagctcattg ccgatagtga gactcctacc attcagaagg gatcttacac ctttgtgcct 1440
tggctgctgt ctttcaaacg gggctctgct ctggaggaaa aggagaacaa aattctggtc 1500
aaagagactg gctacttctt catctacggc caggtgctgt acaccgacaa aacatacgcc 1560
atgggccatc tcattcagcg gaaaaaagtc cacgtgttcg gcgacgaact ctctctcgtg 1620
acactgttcc ggtgtatcca gaacatgccc gagacactgc ccaataatag ctgctactct 1680
gccggcattg ctaaactgga ggagggggac gaactccagc tggctattcc tagggaaaat 1740
gcccagattt ctctcgatgg ggatgtgaca ttcttcgggg ccctcaaact gctgggaggc 1800
ggcggatctg gcggaggcgg gagtcaattc gcagcaggtc cagaagaaac agtcactcaa 1860
gactgcttgc aactgattgc agacagtgaa acaccaacta tacaaaaagg atcttacaca 1920
tttgttccat ggcttctcag ctttaaaagg ggaagtgccc tagaagaaaa agagaataaa 1980
atattggtca aagaaactgg ttactttttt atatatggtc aggttttata tactgataag 2040
acctacgcca tgggacatct aattcagagg aagaaggtcc atgtctttgg ggatgaattg 2100
agtctggtga ctttgtttcg atgtattcaa aatatgcctg aaacactacc caataattcc 2160
tgctattcag ctggcattgc aaaactggaa gaaggagatg aactccaact tgcaatacca 2220
agagaaaatg cacaaatatc actggatgga gatgtcacat tttttggtgc attgaaactg 2280
ctgtga 2286
<210> 111
<211> 2169
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG2-VH-scFv:CD20
<400> 111
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatcctcg agtgctagca ccaagggccc atcggtcttc 480
cccctggcgc cctgctccag gagcacctcc gagagcacag cggccctggg ctgcctggtc 540
aaggactact tccccgaacc ggtgacggtg tcgtggaact caggcgctct gaccagcggc 600
gtgcacacct tcccagctgt cctacagtcc tcaggactct actccctcag cagcgtggtg 660
accgtgccct ccagcaactt cggcacccag acctacacct gcaacgtaga tcacaagccc 720
agcaacacca aggtggacaa gacagttgag cgcaaatgtt gtgtcgagtg cccaccgtgc 780
ccagcaccac ctgtggcagg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 840
ctcatgatct cccggacccc tgaggtcacg tgcgtggtgg tggacgtgag ccacgaagac 900
cccgaggtcc agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 960
ccacgggagg agcagttcaa cagcacgttc cgtgtggtca gcgtcctcac cgttgtgcac 1020
caggactggc tgaacggcaa ggagtacaag tgcaaggtct ccaacaaagg cctcccagcc 1080
cccatcgaga aaaccatctc caaaaccaaa gggcagcccc gagaaccaca ggtgtacacc 1140
ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg cctggtcaaa 1200
ggcttctacc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 1260
tacaagacca cgcctcccat gctggactcc gacggctcct tcttcctcta cagcaagctc 1320
accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 1380
gctctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa agaattccag 1440
gtacaactgc agcagcctgg ggctgagctg gtgaagcctg gggcctcagt gaagatgtcc 1500
tgcaaggctt ctggctacac atttaccagt tacaatatgc actgggtaaa acagacacct 1560
ggtcggggcc tggaatggat tggagctatt tatcccggaa atggtgatac ttcctacaat 1620
cagaagttca aaggcaaggc cacattgact gcagacaaat cctccagcac agcctacatg 1680
cagctcagca gcctgacatc tgaggactct gcggtctatt actgtgcaag atcgacttac 1740
tacggcggtg actggtactt caatgtctgg ggcgcaggga ccacggtcac cgtctcttca 1800
ggaggaggcg gatccggcgg aggcggaagc ggtggcggag gctctcaaat tgttctctcc 1860
cagtctccag caatcctgtc tgcatctcca ggggagaagg tcacaatgac ttgcagggcc 1920
agctcaagtg taagttacat ccactggttc cagcagaagc caggatcctc ccccaaaccc 1980
tggatttatg ccacatccaa cctggcttct ggagtccctg ttcgcttcag tggcagtggg 2040
tctgggactt cttactctct cacaatcagc agagtggagg ctgaagatgc tgccacttat 2100
tactgccagc agtggactag taacccaccc acgttcggag gggggaccaa gctggaaatc 2160
aaacgttaa 2169
<210> 112
<211> 2196
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(1F6)
<400> 112
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc agattcagct cgtccagtcc ggacctgaag tgaaaaaacc tggcgaaacc 1500
gtgaaaattt cctgtaaggc ctctggctac acctttacca actacggcat gaactgggtc 1560
aaacaggctc ctgggaaggg cctgaaatgg atgggatgga tcaacaccta caccggcgaa 1620
ccaacatacg ccgatgcctt taagggacgc tttgccttct ctctggaaac ttccgcctct 1680
actgcttacc tccagatcaa taacctcaaa aacgaggaca ccgccactta cttttgtgct 1740
cgggattacg gggactacgg gatggattac tggggacagg gaacatctgt gaccgtgtct 1800
agcgcttcta caaaggggcc taaactggag gagggcgagt ttagcgaggc tagagtggat 1860
atcgtgctca cacagtctcc cgcttctctg gctgtctcac tgggccagcg agcaacaatc 1920
tcttgtcggg cttccaaatc cgtgtctact agcggctact cttttatgca ctggtaccag 1980
cagaaacctg ggcagcctcc aaaactgctc atctacctgg cttcaaacct cgaatccgga 2040
gtgcctgctc gattttctgg ctctggctcc gggaccgact ttacactgaa cattcatcct 2100
gtcgaggagg aggacgctgc cacatactac tgtcagcatt ctagggaggt gccatggaca 2160
tttggcgggg gaacaaaact ggaaatcaaa cggtaa 2196
<210> 113
<211> 2187
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TNFR2(C4)-IgG1(N297A)-VH-scFv:CD70(2H5)
<400> 113
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attgcaggtt cagctgctgc agtctggacc tgagctggtg 120
aagcctgggg cttcagtgaa gttgtcctgc aaggcttctg gttatagttt cacaagttac 180
gatattaact gggtgaagca gaggcctgga cagggacttg agtgggttgg atggatttat 240
cctagagatg gtgatactaa gtacaatgag aaattcaagg gcaaggccat attgactgta 300
gacacatcct ccaacacagc gtacatgaac ctccacagcc tgacatctga ggactctgcg 360
gtctatttct gtgcaagact aactgggccc tactggtact tcgatgtctg gggcacaggg 420
accacggtca ccgtctcctc aggatccagc agcgcctcta caaagggccc cagcgtgttc 480
cctctggccc ctagcagcaa gagcacatct ggcggaacag ccgccctggg ctgcctcgtg 540
aaggactact ttcccgagcc cgtgaccgtg tcctggaact ctggcgctct gacaagcggc 600
gtgcacacct ttccagccgt gctgcagagc agcggcctgt actctctgag cagcgtcgtg 660
acagtgccca gcagctctct gggcacccag acctacatct gcaacgtgaa ccacaagccc 720
agcaacacca aggtggacaa gaaggtggaa cccaagagct gcgacaagac ccacacctgt 780
cccccttgtc ctgcccccga actgctggga ggcccttccg tgttcctgtt ccccccaaag 840
cccaaggaca ccctgatgat cagccggacc cccgaagtga cctgcgtggt ggtggatgtg 900
tcccacgagg accctgaagt gaagtttaat tggtacgtgg acggcgtgga agtgcacaac 960
gccaagacca agcctagaga ggaacagtac gccagcacct accgggtggt gtccgtgctg 1020
acagtgctgc accaggactg gctgaacggc aaagagtaca agtgcaaggt gtccaacaag 1080
gccctgcctg cccccatcga gaaaaccatc agcaaggcca agggccagcc ccgcgaaccc 1140
caggtgtaca cactgccccc aagcagggac gagctgacca agaaccaggt gtccctgacc 1200
tgtctcgtga aaggcttcta ccccagcgat atcgccgtgg aatgggagag caacggccag 1260
cccgagaaca actacaagac caccccccct gtgctggaca gcgacggctc attcttcctg 1320
tacagcaagc tgaccgtgga caagtcccgg tggcagcagg gcaacgtgtt cagctgcagc 1380
gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc 1440
aaggaattcc aggtgcagct ggtggaatct ggcggcggag tggtgcagcc tggcagaagc 1500
ctgagactga gctgtgccgc cagcggcttc accttcagca gctacatcat gcactgggtg 1560
cgccaggccc ctggcaaggg actggaatgg gtggccgtga tcagctacga cggccggaac 1620
aagtactacg ccgacagcgt gaagggccgg ttcaccatct cccgggacaa cagcaagaac 1680
accctgtacc tgcagatgaa cagcctgcgg gccgaggaca ccgccgtgta ctactgtgcc 1740
agagacaccg acggctacga cttcgactat tggggccagg gcaccctcgt gaccgtgtct 1800
agcggaggcg gaggatctgg cggaggggga tcaggcgggg gaggctctga aatcgtgctg 1860
acacagagcc ccgccaccct gtcactgtct ccaggcgaaa gagccaccct gagctgcaga 1920
gccagccaga gcgtgtccag ctacctggcc tggtatcagc agaagcccgg acaggccccc 1980
agactgctga tctacgacgc cagcaatcgg gccacaggca tccctgccag attttccggc 2040
tctggcagcg gcaccgactt caccctgaca atcagcagcc tggaacccga ggactttgcc 2100
gtgtattatt gccagcagcg gaccaactgg cccctgacct ttggcggagg caccaaggtg 2160
gaaatcaagg ccagcaccaa gggctaa 2187
<210> 114
<211> 2211
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK4)-IgG1(N297A)-VH-scFv:CD20
<400> 114
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatccagc 480
agcgcctcta caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgaccgtg 600
tcctggaact ctggcgctct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg acagtgccca gcagctctct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgcccccga actgctggga 840
ggcccttccg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 900
cccgaagtga cctgcgtggt ggtggatgtg tcccacgagg accctgaagt gaagtttaat 960
tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcctagaga ggaacagtac 1020
gccagcacct accgggtggt gtccgtgctg acagtgctgc accaggactg gctgaacggc 1080
aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 1140
agcaaggcca agggccagcc ccgcgaaccc caggtgtaca cactgccccc aagcagggac 1200
gagctgacca agaaccaggt gtccctgacc tgtctcgtga aaggcttcta ccccagcgat 1260
atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 1320
gtgctggaca gcgacggctc attcttcctg tacagcaagc tgaccgtgga caagtcccgg 1380
tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1440
acccagaagt ccctgagcct gagccccggc aagctcgagc aggtacaact gcagcagcct 1500
ggggctgagc tggtgaagcc tggggcctca gtgaagatgt cctgcaaggc ttctggctac 1560
acatttacca gttacaatat gcactgggta aaacagacac ctggtcgggg cctggaatgg 1620
attggagcta tttatcccgg aaatggtgat acttcctaca atcagaagtt caaaggcaag 1680
gccacattga ctgcagacaa atcctccagc acagcctaca tgcagctcag cagcctgaca 1740
tctgaggact ctgcggtcta ttactgtgca agatcgactt actacggcgg tgactggtac 1800
ttcaatgtct ggggcgcagg gaccacggtc accgtctctt caggaggagg cggatccggc 1860
ggaggcggaa gcggtggcgg aggctctcaa attgttctct cccagtctcc agcaatcctg 1920
tctgcatctc caggggagaa ggtcacaatg acttgcaggg ccagctcaag tgtaagttac 1980
atccactggt tccagcagaa gccaggatcc tcccccaaac cctggattta tgccacatcc 2040
aacctggctt ctggagtccc tgttcgcttc agtggcagtg ggtctgggac ttcttactct 2100
ctcacaatca gcagagtgga ggctgaagat gctgccactt attactgcca gcagtggact 2160
agtaacccac ccacgttcgg aggggggacc aagctggaaa tcaaacgtta a 2211
<210> 115
<211> 774
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK4)-VL
<400> 115
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
gtgatgactc agagccccgc cttcctgtct gtgacccctg gcgagaaagt gaccatcacc 180
tgtagagcca gccagaccat cagcgactac ctgcactggt atcagcagaa gcccgatcag 240
gcccctaagc tgctgattaa gtacgcctct cagagcatca gcggcatccc cagcagattt 300
tctggcagcg gctctggcac cgacttcacc tttaccatca gctccctgga agccgaggat 360
gccgccacct actattgtca ggacggccac agcttccctc caacctttgg acagggcaca 420
aagctggaaa tcaagggatc cgaaatcaag cgtacggtgg ccgctcccag cgtgttcatc 480
ttcccaccta gcgacgagca gctgaagtcc ggcacagcct ctgtcgtgtg cctgctgaac 540
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaatgccct gcagagcggc 600
aacagccagg aaagcgtgac cgagcaggac agcaaggact ccacctacag cctgagcagc 660
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaagtgacc 720
caccagggcc tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaa 774
<210> 116
<211> 2250
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD95(E09)-IgG1(N297A)-VH-scFv:CD20
<220>
<221> misc_feature
<222> (1581)..(1581)
<223> n is a, c, g, or t
<400> 116
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagctg 120
cagctgcagg aatctggccc tggcctcgtg aagcccagcg agacactgag cctgacctgt 180
accgtgtccg gcgccagcat cagcgccaac agctactatg gcgtgtgggt gcgccagagc 240
cctggcaagg gactggaatg ggtgggatct atcgcctacc ggggcaacag caacagcggc 300
agcacctact acaaccccag cctgaagtcc cgggccaccg tgtctgtgga caccagcaag 360
aaccaggtgt ccctgcggct gacctctgtg acagccgccg ataccgccct gtactactgc 420
gccagaaggc agctgctgga cgacggcaca ggatatcagt gggccgcctt cgatgtgtgg 480
ggccagggaa caatggtcac cgtgtcctcc ggatccagca gcgcctctac aaagggcccc 540
agcgtgttcc ctctggcccc tagcagcaag agcacatctg gcggaacagc cgccctgggc 600
tgcctcgtga aggactactt tcccgagccc gtgaccgtgt cctggaactc tggcgctctg 660
acaagcggcg tgcacacctt tccagccgtg ctgcagagca gcggcctgta ctctctgagc 720
agcgtcgtga cagtgcccag cagctctctg ggcacccaga cctacatctg caacgtgaac 780
cacaagccca gcaacaccaa ggtggacaag aaggtggaac ccaagagctg cgacaagacc 840
cacacctgtc ccccttgtcc tgcccccgaa ctgctgggag gcccttccgt gttcctgttc 900
cccccaaagc ccaaggacac cctgatgatc agccggaccc ccgaagtgac ctgcgtggtg 960
gtggatgtgt cccacgagga ccctgaagtg aagtttaatt ggtacgtgga cggcgtggaa 1020
gtgcacaacg ccaagaccaa gcctagagag gaacagtacg ccagcaccta ccgggtggtg 1080
tccgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140
tccaacaagg ccctgcctgc ccccatcgag aaaaccatca gcaaggccaa gggccagccc 1200
cgcgaacccc aggtgtacac actgccccca agcagggacg agctgaccaa gaaccaggtg 1260
tccctgacct gtctcgtgaa aggcttctac cccagcgata tcgccgtgga atgggagagc 1320
aacggccagc ccgagaacaa ctacaagacc accccccctg tgctggacag cgacggctca 1380
ttcttcctgt acagcaagct gaccgtggac aagtcccggt ggcagcaggg caacgtgttc 1440
agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg 1500
agccccggca aggaattcca ggtacaactg cagcagcctg gggctgagct ggtgaagcct 1560
ggggcctcag tgaagatgtc ntgcaaggct tctggctaca catttaccag ttacaatatg 1620
cactgggtaa aacagacacc tggtcggggc ctggaatgga ttggagctat ttatcccgga 1680
aatggtgata cttcctacaa tcagaagttc aaaggcaagg ccacattgac tgcagacaaa 1740
tcctccagca cagcctacat gcagctcagc agcctgacat ctgaggactc tgcggtctat 1800
tactgtgcaa gatcgactta ctacggcggt gactggtact tcaatgtctg gggcgcaggg 1860
accacggtca ccgtctcttc aggaggaggc ggatccggcg gaggcggaag cggtggcgga 1920
ggctctcaaa ttgttctctc ccagtctcca gcaatcctgt ctgcatctcc aggggagaag 1980
gtcacaatga cttgcagggc cagctcaagt gtaagttaca tccactggtt ccagcagaag 2040
ccaggatcct cccccaaacc ctggatttat gccacatcca acctggcttc tggagtccct 2100
gttcgcttca gtggcagtgg gtctgggact tcttactctc tcacaatcag cagagtggag 2160
gctgaagatg ctgccactta ttactgccag cagtggacta gtaacccacc cacgttcgga 2220
ggggggacca agctggaaat caaacgttaa 2250
<210> 117
<211> 268
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 108所示的蛋白质构建体
<400> 117
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
260 265
<210> 118
<211> 741
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 109所示的蛋白质构建体
<400> 118
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Leu Gln Gln Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly
50 55 60
Tyr Ser Ile Thr Thr Asn Tyr Asn Trp Asn Trp Ile Arg Gln Phe Pro
65 70 75 80
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Arg Tyr Asp Gly Thr Ser
85 90 95
Glu Tyr Thr Pro Ser Leu Lys Asn Arg Val Ser Ile Thr Arg Asp Thr
100 105 110
Ser Met Asn Gln Phe Phe Leu Arg Leu Thr Ser Val Thr Pro Glu Asp
115 120 125
Thr Ala Thr Tyr Tyr Cys Ala Arg Leu Asp Tyr Trp Gly Gln Gly Thr
130 135 140
Leu Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro
145 150 155 160
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
165 170 175
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
180 185 190
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
195 200 205
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
210 215 220
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
225 230 235 240
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
245 250 255
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Leu Glu Gly Pro
260 265 270
Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu
275 280 285
Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu
290 295 300
Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu
305 310 315 320
Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr
325 330 335
Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His
340 345 350
Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln
355 360 365
Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile
370 375 380
Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu
385 390 395 400
Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu
405 410 415
Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Pro Glu
420 425 430
Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp Ser Glu Thr
435 440 445
Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp Leu Leu Ser
450 455 460
Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys Ile Leu Val
465 470 475 480
Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu Tyr Thr Asp
485 490 495
Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys Val His Val
500 505 510
Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys Ile Gln Asn
515 520 525
Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala Gly Ile Ala
530 535 540
Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro Arg Glu Asn
545 550 555 560
Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly Ala Leu Lys
565 570 575
Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Phe Ala Ala
580 585 590
Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala Asp
595 600 605
Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro Trp
610 615 620
Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn Lys
625 630 635 640
Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val Leu
645 650 655
Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys Lys
660 665 670
Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg Cys
675 680 685
Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser Ala
690 695 700
Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile Pro
705 710 715 720
Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe Gly
725 730 735
Ala Leu Lys Leu Leu
740
<210> 119
<211> 761
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 110所示的蛋白质构建体
<400> 119
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Leu Glu Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile
290 295 300
Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val
305 310 315 320
Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu
325 330 335
Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln
340 345 350
Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg
355 360 365
Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe
370 375 380
Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr
385 390 395 400
Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala
405 410 415
Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe
420 425 430
Phe Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly
435 440 445
Ser Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln Leu Ile Ala
450 455 460
Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr Phe Val Pro
465 470 475 480
Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu Lys Glu Asn
485 490 495
Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr Gly Gln Val
500 505 510
Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile Gln Arg Lys
515 520 525
Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr Leu Phe Arg
530 535 540
Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser Cys Tyr Ser
545 550 555 560
Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln Leu Ala Ile
565 570 575
Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val Thr Phe Phe
580 585 590
Gly Ala Leu Lys Leu Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
595 600 605
Gln Phe Ala Ala Gly Pro Glu Glu Thr Val Thr Gln Asp Cys Leu Gln
610 615 620
Leu Ile Ala Asp Ser Glu Thr Pro Thr Ile Gln Lys Gly Ser Tyr Thr
625 630 635 640
Phe Val Pro Trp Leu Leu Ser Phe Lys Arg Gly Ser Ala Leu Glu Glu
645 650 655
Lys Glu Asn Lys Ile Leu Val Lys Glu Thr Gly Tyr Phe Phe Ile Tyr
660 665 670
Gly Gln Val Leu Tyr Thr Asp Lys Thr Tyr Ala Met Gly His Leu Ile
675 680 685
Gln Arg Lys Lys Val His Val Phe Gly Asp Glu Leu Ser Leu Val Thr
690 695 700
Leu Phe Arg Cys Ile Gln Asn Met Pro Glu Thr Leu Pro Asn Asn Ser
705 710 715 720
Cys Tyr Ser Ala Gly Ile Ala Lys Leu Glu Glu Gly Asp Glu Leu Gln
725 730 735
Leu Ala Ile Pro Arg Glu Asn Ala Gln Ile Ser Leu Asp Gly Asp Val
740 745 750
Thr Phe Phe Gly Ala Leu Lys Leu Leu
755 760
<210> 120
<211> 722
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 111所示的蛋白质构建体
<400> 120
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu
245 250 255
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
260 265 270
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
275 280 285
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln
290 295 300
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
305 310 315 320
Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu
325 330 335
Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
340 345 350
Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
355 360 365
Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
370 375 380
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
385 390 395 400
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
405 410 415
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly
420 425 430
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
435 440 445
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
450 455 460
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln
465 470 475 480
Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser
485 490 495
Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn
500 505 510
Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly
515 520 525
Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys
530 535 540
Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met
545 550 555 560
Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala
565 570 575
Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala
580 585 590
Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
595 600 605
Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala
610 615 620
Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala
625 630 635 640
Ser Ser Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser
645 650 655
Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val
660 665 670
Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr
675 680 685
Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
690 695 700
Trp Thr Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile
705 710 715 720
Lys Arg
<210> 121
<211> 731
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 112所示的蛋白质构建体
<400> 121
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Val Lys Lys
485 490 495
Pro Gly Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe
500 505 510
Thr Asn Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu
515 520 525
Lys Trp Met Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala
530 535 540
Asp Ala Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser
545 550 555 560
Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr
565 570 575
Tyr Phe Cys Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly
580 585 590
Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys
595 600 605
Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg Val Asp Ile Val Leu Thr
610 615 620
Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile
625 630 635 640
Ser Cys Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met
645 650 655
His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr
660 665 670
Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
675 680 685
Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu
690 695 700
Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg Glu Val Pro Trp Thr
705 710 715 720
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
725 730
<210> 122
<211> 728
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 113所示的蛋白质构建体
<400> 122
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Gln Val Gln Leu
20 25 30
Leu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu
35 40 45
Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr Asp Ile Asn Trp
50 55 60
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Val Gly Trp Ile Tyr
65 70 75 80
Pro Arg Asp Gly Asp Thr Lys Tyr Asn Glu Lys Phe Lys Gly Lys Ala
85 90 95
Ile Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr Met Asn Leu His
100 105 110
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Leu Thr
115 120 125
Gly Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr Val Thr
130 135 140
Val Ser Ser Gly Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
145 150 155 160
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
165 170 175
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
180 185 190
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
195 200 205
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
210 215 220
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
225 230 235 240
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
245 250 255
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
260 265 270
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
275 280 285
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
290 295 300
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
305 310 315 320
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val
325 330 335
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
340 345 350
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
355 360 365
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
370 375 380
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
385 390 395 400
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
405 410 415
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
420 425 430
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
435 440 445
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
450 455 460
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
465 470 475 480
Lys Glu Phe Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
485 490 495
Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
500 505 510
Ser Ser Tyr Ile Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
515 520 525
Glu Trp Val Ala Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala
530 535 540
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
545 550 555 560
Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
565 570 575
Tyr Tyr Cys Ala Arg Asp Thr Asp Gly Tyr Asp Phe Asp Tyr Trp Gly
580 585 590
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
595 600 605
Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro
610 615 620
Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
625 630 635 640
Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
645 650 655
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr
660 665 670
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
675 680 685
Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
690 695 700
Gln Gln Arg Thr Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val
705 710 715 720
Glu Ile Lys Ala Ser Thr Lys Gly
725
<210> 123
<211> 736
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 114所示的蛋白质构建体
<400> 123
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
275 280 285
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
290 295 300
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
305 310 315 320
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
325 330 335
Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
340 345 350
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
355 360 365
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
370 375 380
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
385 390 395 400
Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
405 410 415
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
420 425 430
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
435 440 445
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
450 455 460
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
465 470 475 480
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Gln Val Gln
485 490 495
Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys
500 505 510
Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His
515 520 525
Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile
530 535 540
Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys
545 550 555 560
Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu
565 570 575
Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser
580 585 590
Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr
595 600 605
Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
610 615 620
Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu
625 630 635 640
Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser
645 650 655
Ser Val Ser Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro
660 665 670
Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val
675 680 685
Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser
690 695 700
Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr
705 710 715 720
Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
725 730 735
<210> 124
<211> 257
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 115所示的蛋白质构建体
<400> 124
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Val Met Thr Gln Ser Pro Ala Phe
35 40 45
Leu Ser Val Thr Pro Gly Glu Lys Val Thr Ile Thr Cys Arg Ala Ser
50 55 60
Gln Thr Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Pro Asp Gln
65 70 75 80
Ala Pro Lys Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile
85 90 95
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr
100 105 110
Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asp
115 120 125
Gly His Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
130 135 140
Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
145 150 155 160
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
165 170 175
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
180 185 190
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
195 200 205
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
210 215 220
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
225 230 235 240
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
245 250 255
Cys
<210> 125
<211> 749
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 116所示的蛋白质构建体
<400> 125
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Ala Ser Ile Ser Ala Asn Ser Tyr Tyr Gly Val Trp Val Arg Gln Ser
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Val Gly Ser Ile Ala Tyr Arg Gly Asn
85 90 95
Ser Asn Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Ala
100 105 110
Thr Val Ser Val Asp Thr Ser Lys Asn Gln Val Ser Leu Arg Leu Thr
115 120 125
Ser Val Thr Ala Ala Asp Thr Ala Leu Tyr Tyr Cys Ala Arg Arg Gln
130 135 140
Leu Leu Asp Asp Gly Thr Gly Tyr Gln Trp Ala Ala Phe Asp Val Trp
145 150 155 160
Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser Ser Ser Ala Ser
165 170 175
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
180 185 190
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
195 200 205
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
210 215 220
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
225 230 235 240
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
245 250 255
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
260 265 270
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
275 280 285
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
290 295 300
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
305 310 315 320
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
325 330 335
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
340 345 350
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
355 360 365
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
370 375 380
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
385 390 395 400
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
405 410 415
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
420 425 430
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
435 440 445
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
450 455 460
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
465 470 475 480
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
485 490 495
Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln
500 505 510
Pro Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys
515 520 525
Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys
530 535 540
Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly
545 550 555 560
Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu
565 570 575
Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu
580 585 590
Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr
595 600 605
Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr
610 615 620
Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
625 630 635 640
Gly Ser Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser
645 650 655
Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser
660 665 670
Tyr Ile His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp
675 680 685
Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser
690 695 700
Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu
705 710 715 720
Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro
725 730 735
Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
740 745
<210> 126
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR1-H
<400> 126
Tyr Thr Phe Ser Ser Tyr Trp Met His
1 5
<210> 127
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR2-H
<400> 127
Glu Ile Asn Pro Gly Asn Gly His Thr Asn Tyr Asn Glu Lys Phe Lys
1 5 10 15
Ser
<210> 128
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR3-H
<400> 128
Ser Phe Thr Thr Ala Arg Ala Phe Ala Tyr
1 5 10
<210> 129
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR1-L
<400> 129
Arg Ala Ser Gln Thr Ile Ser Asp Tyr Leu His
1 5 10
<210> 130
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR2-L
<400> 130
Leu Ala Ser Gln Ser Ile Ser
1 5
<210> 131
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> HBBK4 CDR3-L
<400> 131
Gln Asp Gly His Ser Phe Pro Pro Thr
1 5
<210> 132
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR1-H
<400> 132
Gly Ser Phe Ser Gly Tyr Tyr Trp Ser
1 5
<210> 133
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR2-H
<400> 133
Glu Ile Asn His Gly Gly Tyr Val Thr Tyr Asn Pro Ser Leu Glu Ser
1 5 10 15
<210> 134
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR3-H
<400> 134
Asp Tyr Gly Pro Gly Asn Tyr Asp Trp Tyr Phe Asp Leu
1 5 10
<210> 135
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR1-L
<400> 135
Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
1 5 10
<210> 136
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR2-L
<400> 136
Asp Ala Ser Asn Arg Ala Thr
1 5
<210> 137
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> URELUMAB CDR3-L
<400> 137
Gln Gln Arg Ser Asn Trp Pro Pro Ala Leu Thr
1 5 10
<210> 138
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR1-H
<400> 138
Tyr Ser Phe Ser Thr Tyr Trp Ile Ser
1 5
<210> 139
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR2-H
<400> 139
Lys Ile Tyr Pro Gly Asp Ser Tyr Thr Asn Tyr Ser Pro Ser Phe Gln
1 5 10 15
Gly
<210> 140
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR3-H
<400> 140
Gly Tyr Gly Ile Phe Asp Tyr
1 5
<210> 141
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR1-L
<400> 141
Ser Gly Asp Asn Ile Gly Asp Gln Tyr Ala His
1 5 10
<210> 142
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR2-L
<400> 142
Gln Asp Lys Asn Arg Pro Ser
1 5
<210> 143
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> UTOMILUMAB CDR3-L
<400> 143
Ala Thr Tyr Thr Gly Phe Gly Ser Leu Ala Val
1 5 10
<210> 144
<211> 248
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 121所示的蛋白质构建体的scFv:CD70(1F6)锚定结构域
<400> 144
Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30
Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met
35 40 45
Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe
50 55 60
Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80
Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys
85 90 95
Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys Leu Glu Glu
115 120 125
Gly Glu Phe Ser Glu Ala Arg Val Asp Ile Val Leu Thr Gln Ser Pro
130 135 140
Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg
145 150 155 160
Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His Trp Tyr
165 170 175
Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser
180 185 190
Asn Leu Glu Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
195 200 205
Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala
210 215 220
Thr Tyr Tyr Cys Gln His Ser Arg Glu Val Pro Trp Thr Phe Gly Gly
225 230 235 240
Gly Thr Lys Leu Glu Ile Lys Arg
245
<210> 145
<211> 245
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 122所示的蛋白质构建体的scFv:CD70(2H5)锚定结构域
<400> 145
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Arg Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Thr Asp Gly Tyr Asp Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln
145 150 155 160
Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
165 170 175
Pro Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro
180 185 190
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
195 200 205
Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg
210 215 220
Thr Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
225 230 235 240
Ala Ser Thr Lys Gly
245
<210> 146
<211> 2247
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TRAILR2(Cona)-IgG1(N297A)-VH-scFv:CD70(9G2)
<400> 146
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctgcagg aatctggccc tggcctcgtg aagcctagcc agaccctgag cctgacctgt 180
accgtgtctg gcggcagcat cagcagcggc gactacttct ggtcctggat cagacagctg 240
cccggcaagg gcctggaatg gatcggccac atccacaaca gcggcaccac ctactacaac 300
cccagcctga agtccagagt gaccatcagc gtggacacca gcaagaagca gttcagcctg 360
cggctgagca gcgtgacagc cgccgataca gccgtgtact actgcgccag agacagaggc 420
ggcgattact actacggcat ggacgtgtgg ggccagggca ccaccgtgac cgtgtctagc 480
agatccagca gcgcctctac aaagggcccc agcgtgttcc ctctggcccc tagcagcaag 540
agcacatctg gcggaacagc cgccctgggc tgcctcgtga aggactactt tcccgagccc 600
gtgaccgtgt cctggaactc tggcgctctg acaagcggcg tgcacacctt tccagccgtg 660
ctgcagagca gcggcctgta ctctctgagc agcgtcgtga cagtgcccag cagctctctg 720
ggcacccaga cctacatctg caacgtgaac cacaagccca gcaacaccaa ggtggacaag 780
aaggtggaac ccaagagctg cgacaagacc cacacctgtc ccccttgtcc tgcccccgaa 840
ctgctgggag gcccttccgt gttcctgttc cccccaaagc ccaaggacac cctgatgatc 900
agccggaccc ccgaagtgac ctgcgtggtg gtggatgtgt cccacgagga ccctgaagtg 960
aagtttaatt ggtacgtgga cggcgtggaa gtgcacaacg ccaagaccaa gcctagagag 1020
gaacagtacg ccagcaccta ccgggtggtg tccgtgctga cagtgctgca ccaggactgg 1080
ctgaacggca aagagtacaa gtgcaaggtg tccaacaagg ccctgcctgc ccccatcgag 1140
aaaaccatca gcaaggccaa gggccagccc cgcgaacccc aggtgtacac actgccccca 1200
agcagggacg agctgaccaa gaaccaggtg tccctgacct gtctcgtgaa aggcttctac 1260
cccagcgata tcgccgtgga atgggagagc aacggccagc ccgagaacaa ctacaagacc 1320
accccccctg tgctggacag cgacggctca ttcttcctgt acagcaagct gaccgtggac 1380
aagtcccggt ggcagcaggg caacgtgttc agctgcagcg tgatgcacga ggccctgcac 1440
aaccactaca cccagaagtc cctgagcccc ggcaagctcg agcaggtgca gctggtggaa 1500
tctggcggcg gactgatgca gcctggcggc tctctgagac tgagctgtgc cgccagcggc 1560
ttcaccttta gcagcagcgc catgagctgg gtgcgccagg ctcctggaaa gggcctggaa 1620
tgggtgtcca gcatctacag cgacagcagc tacacctact acgccgacag cgtgaagtcc 1680
cggttcacca tcagcaccga caacgccaag aacaccctgt acctgcagat gaacagcctg 1740
aagcccgacg acaccgccgt gtactactgt gccggcagca gcgattacga gggcagcttt 1800
gcctcttggg gccagggcac acaagtgacc gtgtcctcca gatctagcac caagggcccc 1860
aagctggaag agggcgagtt cagcgaggcc caattgcaga gcgtcgtgac ccagcctcct 1920
agcctgtctg cctctcctgg aagcagcgtg cggctgacct gtacactgag cagcggcaac 1980
agcgtgggca actacgacat cagctggtat cagcagaagg ccggcagccc ccccagatac 2040
ctgctgtact actacagcga ttccgtgaag caccagggca gcggcgtgcc cagcagattt 2100
tccggaagct ctgacgccag cgccaacgcc ggactgctgc tgatttctgg cctgcagcct 2160
gaggacgagg ccgactacta ctgcagcgcc tacaagagcg gcagccacgt gttcggcgga 2220
ggcaccaaac tgacagtgct gggctaa 2247
<210> 147
<211> 768
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗TRAILR2(Cona)-VL
<400> 147
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggagatc 120
gtgctgaccc agagccctgg caccctgtca ctgtctccag gcgagagagc caccctgagc 180
tgtagagcca gccagggcat cagccggtct tacctggcct ggtatcagca gaagcccggc 240
caggctccta gcctgctgat ctacggcgcc agcagcagag ccaccggcat ccccgataga 300
ttttccggca gcggctccgg caccgacttc accctgacaa tcagcagact ggaacccgag 360
gacttcgccg tgtattattg ccagcagttc ggcagcagcc cctggacctt tggccaggga 420
acaaaagtgg gatccgaaat caagcgtacg gtggccgctc ccagcgtgtt catcttccca 480
cctagcgacg agcagctgaa gtccggcaca gcctctgtcg tgtgcctgct gaacaacttc 540
tacccccgcg aggccaaggt gcagtggaag gtggacaatg ccctgcagag cggcaacagc 600
caggaaagcg tgaccgagca ggacagcaag gactccacct acagcctgag cagcaccctg 660
accctgagca aggccgacta cgagaagcac aaggtgtacg cctgcgaagt gacccaccag 720
ggcctgtcta gccccgtgac caagagcttc aaccggggcg agtgctaa 768
<210> 148
<211> 2226
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
<400> 148
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctggttc agtctggcgc cgaagtgaaa aagcctggcg cctctgtgaa ggtgtcctgt 180
acagccagcg gcttcaacat caaggactac tacgtgcact gggtcaagca ggcccctgga 240
caaggactgg aatggatggg cagaatcgac cccgaggacg gcgactctaa gtacgcccct 300
aagttccagg gcaaagccac catgaccgcc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcaccaccag ctactatgtg 420
ggcacctacg gctattgggg ccagggcaca ctggtcaccg tgtccagcag atccagcagc 480
gcctctacaa agggccccag cgtgttccct ctggccccta gcagcaagag cacatctggc 540
ggaacagccg ccctgggctg cctcgtgaag gactactttc ccgagcccgt gaccgtgtcc 600
tggaactctg gcgctctgac aagcggcgtg cacacctttc cagccgtgct gcagagcagc 660
ggcctgtact ctctgagcag cgtcgtgaca gtgcccagca gctctctggg cacccagacc 720
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggtggaaccc 780
aagagctgcg acaagaccca cacctgtccc ccttgtcctg cccccgaact gctgggaggc 840
ccttccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc 900
gaagtgacct gcgtggtggt ggatgtgtcc cacgaggacc ctgaagtgaa gtttaattgg 960
tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ctagagagga acagtacgcc 1020
agcacctacc gggtggtgtc cgtgctgaca gtgctgcacc aggactggct gaacggcaaa 1080
gagtacaagt gcaaggtgtc caacaaggcc ctgcctgccc ccatcgagaa aaccatcagc 1140
aaggccaagg gccagccccg cgaaccccag gtgtacacac tgcccccaag cagggacgag 1200
ctgaccaaga accaggtgtc cctgacctgt ctcgtgaaag gcttctaccc cagcgatatc 1260
gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg 1320
ctggacagcg acggctcatt cttcctgtac agcaagctga ccgtggacaa gtcccggtgg 1380
cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1440
cagaagtccc tgagcctgag ccccggcaag ctcgaggagg tgcagctgct ggaatctggc 1500
ggaggacttg ttcagcctgg cggctctctg agactgtctt gtgccgccag cggcttcacc 1560
ttcagcagct atatcatgat gtgggtccga caggcccctg gcaaaggcct tgaatgggtg 1620
tccagcatct atcccagcgg cggcatcacc ttttacgccg acacagtgaa gggcagattc 1680
accatcagcc gggacaacag caagaacacc ctgtacctgc agatgaacag cctgagagcc 1740
gaggacaccg ccgtgtacta ctgcgccaga atcaagctgg gcaccgtgac caccgtggat 1800
tattggggac agggcaccct ggtcaccgtg tcctccagat cttctacaaa gggccccaag 1860
ctggaagagg gcgagtttag cgaagcccaa ttgcagagcg ccctgacaca gcctgcatcc 1920
gtgtctggat ctccaggcca gagcatcacc atctcttgta ccggcacaag ctccgatgtc 1980
ggcggctaca attacgtgtc ctggtatcag cagcaccccg gcaaggcccc taagctgatg 2040
atctacgacg tgtccaacag accctccggc gtgtccaata gattcagcgg cagcaagagc 2100
ggcaacaccg ccagcctgac aattagcgga ctgcaggccg aggacgaggc cgattactac 2160
tgtagcagct acaccagctc ctccaccaga gtgtttggca ccggcaccaa agtgaccgtg 2220
ctttaa 2226
<210> 149
<211> 771
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-VL
<400> 149
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
cagatgacac agagccccag cagcctgtct gccagcgtgg gagatagagt gaccatcacc 180
tgtagcgcca gcagcagcgt gtcctacatg ctgtggttcc agcagaagcc tggcaaggcc 240
cctaagctgc tgatctacag cacctccaat ctggccagcg gcgtgccaag cagattttct 300
ggctctggca gcggcaccga cttcaccctg accatatcta gcctgcagcc agaggacttc 360
gccacctact actgccagca gcggacattc tacccctaca cctttggcgg aggcaccaag 420
gtggaaatca agggatccga aatcaagcgt acggtggccg ctcccagcgt gttcatcttc 480
ccacctagcg acgagcagct gaagtccggc acagcctctg tcgtgtgcct gctgaacaac 540
ttctaccccc gcgaggccaa ggtgcagtgg aaggtggaca atgccctgca gagcggcaac 600
agccaggaaa gcgtgaccga gcaggacagc aaggactcca cctacagcct gagcagcacc 660
ctgaccctga gcaaggccga ctacgagaag cacaaggtgt acgcctgcga agtgacccac 720
cagggcctgt ctagccccgt gaccaagagc ttcaaccggg gcgagtgcta a 771
<210> 150
<211> 1578
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗CD40(C)-Fab2-scFv:PD-L1(Ave)
<400> 150
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtg 120
cagctggttc agtctggcgc cgaagtgaaa aagcctggcg cctctgtgaa ggtgtcctgt 180
acagccagcg gcttcaacat caaggactac tacgtgcact gggtcaagca ggcccctgga 240
caaggactgg aatggatggg cagaatcgac cccgaggacg gcgactctaa gtacgcccct 300
aagttccagg gcaaagccac catgaccgcc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcaccaccag ctactatgtg 420
ggcacctacg gctattgggg ccagggcaca ctggtcaccg tgtccagcag atcctctagc 480
gccagcacaa agggccccag cgtgttccct ctggccccta gcagcaagag cacatctggc 540
ggaacagccg ccctgggctg cctcgtgaag gactactttc ccgagcccgt gacagtgtcc 600
tggaactctg gcgccctgac aagcggcgtg cacacctttc cagccgtgct gcagagcagc 660
ggcctgtact ctctgagcag cgtcgtgact gtgcccagca gcagcctggg cacccagacc 720
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggtggaaccc 780
aagagctgcg acaagaccca cacctgtccc ccttgtcctg ccctcgagga ggtgcagctg 840
ctggaatctg gcggaggact tgttcagcct ggcggctctc tgagactgtc ttgtgccgcc 900
agcggcttca ccttcagcag ctatatcatg atgtgggtcc gacaggcccc tggcaaaggc 960
cttgaatggg tgtccagcat ctatcccagc ggcggcatca ccttttacgc cgacacagtg 1020
aagggcagat tcaccatcag ccgggacaac agcaagaaca ccctgtacct gcagatgaac 1080
agcctgagag ccgaggacac cgccgtgtac tactgcgcca gaatcaagct gggcaccgtg 1140
accaccgtgg attattgggg acagggcacc ctggtcaccg tgtcctccag atcttctaca 1200
aagggcccca agctggaaga gggcgagttt agcgaagccc aattgcagag cgccctgaca 1260
cagcctgcat ccgtgtctgg atctccaggc cagagcatca ccatctcttg taccggcaca 1320
agctccgatg tcggcggcta caattacgtg tcctggtatc agcagcaccc cggcaaggcc 1380
cctaagctga tgatctacga cgtgtccaac agaccctccg gcgtgtccaa tagattcagc 1440
ggcagcaaga gcggcaacac cgccagcctg acaattagcg gactgcaggc cgaggacgag 1500
gccgattact actgtagcag ctacaccagc tcctccacca gagtgtttgg caccggcacc 1560
aaagtgaccg tgctttaa 1578
<210> 151
<211> 2229
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK)-IgG1(N297A)-VH-scFv:PD-L1(Ave)
<400> 151
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatccagc 480
agcgcctcta caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgaccgtg 600
tcctggaact ctggcgctct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg acagtgccca gcagctctct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgcccccga actgctggga 840
ggcccttccg tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc 900
cccgaagtga cctgcgtggt ggtggatgtg tcccacgagg accctgaagt gaagtttaat 960
tggtacgtgg acggcgtgga agtgcacaac gccaagacca agcctagaga ggaacagtac 1020
gccagcacct accgggtggt gtccgtgctg acagtgctgc accaggactg gctgaacggc 1080
aaagagtaca agtgcaaggt gtccaacaag gccctgcctg cccccatcga gaaaaccatc 1140
agcaaggcca agggccagcc ccgcgaaccc caggtgtaca cactgccccc aagcagggac 1200
gagctgacca agaaccaggt gtccctgacc tgtctcgtga aaggcttcta ccccagcgat 1260
atcgccgtgg aatgggagag caacggccag cccgagaaca actacaagac caccccccct 1320
gtgctggaca gcgacggctc attcttcctg tacagcaagc tgaccgtgga caagtcccgg 1380
tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1440
acccagaagt ccctgagcct gagccccggc aagctcgagg aggtgcagct gctggaatct 1500
ggcggaggac ttgttcagcc tggcggctct ctgagactgt cttgtgccgc cagcggcttc 1560
accttcagca gctatatcat gatgtgggtc cgacaggccc ctggcaaagg ccttgaatgg 1620
gtgtccagca tctatcccag cggcggcatc accttttacg ccgacacagt gaagggcaga 1680
ttcaccatca gccgggacaa cagcaagaac accctgtacc tgcagatgaa cagcctgaga 1740
gccgaggaca ccgccgtgta ctactgcgcc agaatcaagc tgggcaccgt gaccaccgtg 1800
gattattggg gacagggcac cctggtcacc gtgtcctcca gatcttctac aaagggcccc 1860
aagctggaag agggcgagtt tagcgaagcc caattgcaga gcgccctgac acagcctgca 1920
tccgtgtctg gatctccagg ccagagcatc accatctctt gtaccggcac aagctccgat 1980
gtcggcggct acaattacgt gtcctggtat cagcagcacc ccggcaaggc ccctaagctg 2040
atgatctacg acgtgtccaa cagaccctcc ggcgtgtcca atagattcag cggcagcaag 2100
agcggcaaca ccgccagcct gacaattagc ggactgcagg ccgaggacga ggccgattac 2160
tactgtagca gctacaccag ctcctccacc agagtgtttg gcaccggcac caaagtgacc 2220
gtgctttaa 2229
<210> 152
<211> 1581
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗41BB(HBBK)-Fab2-scFv:PD-L1(Ave)
<400> 152
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaagttatt aagcctggcg cctccgtgaa gctgagctgt 180
aaagccagcg gctacacctt cagcagctac tggatgcact gggtccgaca ggctccagga 240
caaggcctgg aatggatcgg cgagatcaac cctggcaacg gccacaccaa ctacaacgag 300
aagttcaaga gccgggccac actgaccggc gataccagca caagcaccgt gtacatggaa 360
ctgagcagcc tgagaagcga ggacaccgcc gtgtactact gcgccagatc ctttaccacc 420
gccagagcct ttgcctattg gggccaggga acactggtca ccgtgtccag cagatcctct 480
agcgccagca caaagggccc cagcgtgttc cctctggccc ctagcagcaa gagcacatct 540
ggcggaacag ccgccctggg ctgcctcgtg aaggactact ttcccgagcc cgtgacagtg 600
tcctggaact ctggcgccct gacaagcggc gtgcacacct ttccagccgt gctgcagagc 660
agcggcctgt actctctgag cagcgtcgtg actgtgccca gcagcagcct gggcacccag 720
acctacatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gaaggtggaa 780
cccaagagct gcgacaagac ccacacctgt cccccttgtc ctgccctcga ggaggtgcag 840
ctgctggaat ctggcggagg acttgttcag cctggcggct ctctgagact gtcttgtgcc 900
gccagcggct tcaccttcag cagctatatc atgatgtggg tccgacaggc ccctggcaaa 960
ggccttgaat gggtgtccag catctatccc agcggcggca tcacctttta cgccgacaca 1020
gtgaagggca gattcaccat cagccgggac aacagcaaga acaccctgta cctgcagatg 1080
aacagcctga gagccgagga caccgccgtg tactactgcg ccagaatcaa gctgggcacc 1140
gtgaccaccg tggattattg gggacagggc accctggtca ccgtgtcctc cagatcttct 1200
acaaagggcc ccaagctgga agagggcgag tttagcgaag cccaattgca gagcgccctg 1260
acacagcctg catccgtgtc tggatctcca ggccagagca tcaccatctc ttgtaccggc 1320
acaagctccg atgtcggcgg ctacaattac gtgtcctggt atcagcagca ccccggcaag 1380
gcccctaagc tgatgatcta cgacgtgtcc aacagaccct ccggcgtgtc caatagattc 1440
agcggcagca agagcggcaa caccgccagc ctgacaatta gcggactgca ggccgaggac 1500
gaggccgatt actactgtag cagctacacc agctcctcca ccagagtgtt tggcaccggc 1560
accaaagtga ccgtgcttta a 1581
<210> 153
<211> 2208
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗PD-L1(Ave)-IgG2-VH-scFv:41BB(HBBK)
<400> 153
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attcgaggtg 120
cagctgctgg aatctggcgg aggacttgtt cagcctggcg gctctctgag actgtcttgt 180
gccgccagcg gcttcacctt cagcagctat atcatgatgt gggtccgaca ggcccctggc 240
aaaggccttg aatgggtgtc cagcatctat cccagcggcg gcatcacctt ttacgccgac 300
acagtgaagg gcagattcac catcagccgg gacaacagca agaacaccct gtacctgcag 360
atgaacagcc tgagagccga ggacaccgcc gtgtactact gcgccagaat caagctgggc 420
accgtgacca ccgtggatta ttggggacag ggcaccctgg tcaccgtgtc ctccagatcc 480
tcgagtgcta gcaccaaggg cccatcggtc ttccccctgg cgccctgctc caggagcacc 540
tccgagagca cagcggccct gggctgcctg gtcaaggact acttccccga accggtgacg 600
gtgtcgtgga actcaggcgc tctgaccagc ggcgtgcaca ccttcccagc tgtcctacag 660
tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcaa cttcggcacc 720
cagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagacagtt 780
gagcgcaaat gttgtgtcga gtgcccaccg tgcccagcac cacctgtggc aggaccgtca 840
gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 900
acgtgcgtgg tggtggacgt gagccacgaa gaccccgagg tccagttcaa ctggtacgtg 960
gacggcgtgg aggtgcataa tgccaagaca aagccacggg aggagcagtt caacagcacg 1020
ttccgtgtgg tcagcgtcct caccgttgtg caccaggact ggctgaacgg caaggagtac 1080
aagtgcaagg tctccaacaa aggcctccca gcccccatcg agaaaaccat ctccaaaacc 1140
aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 1200
aagaaccagg tcagcctgac ctgcctggtc aaaggcttct accccagcga catcgccgtg 1260
gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc catgctggac 1320
tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1380
gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1440
agcctctccc tgtctccggg taaagaattc caggtccagc tgcagcagtc tggcgccgaa 1500
gttattaagc ctggcgcctc cgtgaagctg agctgtaaag ccagcggcta caccttcagc 1560
agctactgga tgcactgggt ccgacaggct ccaggacaag gcctggaatg gatcggcgag 1620
atcaaccctg gcaacggcca caccaactac aacgagaagt tcaagagccg ggccacactg 1680
accggcgata ccagcacaag caccgtgtac atggaactga gcagcctgag aagcgaggac 1740
accgccgtgt actactgcgc cagatccttt accaccgcca gagcctttgc ctattggggc 1800
cagggaacac tggtcaccgt gtccagcaga tctagcacaa agggccccaa gctggaagag 1860
ggcgagttta gcgaggccca attggacatc gtgatgactc agagccccgc cttcctgtct 1920
gtgacccctg gcgagaaagt gaccatcacc tgtagagcca gccagaccat cagcgactac 1980
ctgcactggt atcagcagaa gcccgatcag gcccctaagc tgctgattaa gtacgcctct 2040
cagagcatca gcggcatccc cagcagattt tctggcagcg gctctggcac cgacttcacc 2100
tttaccatca gctccctgga agccgaggat gccgccacct actattgtca ggacggccac 2160
agcttccctc caacctttgg acagggcaca aagctggaaa tcaagtaa 2208
<210> 154
<211> 783
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗PD-L1(Ave)-VL
<400> 154
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attgcagagc 120
gccctgacac agcctgcatc cgtgtctgga tctccaggcc agagcatcac catctcttgt 180
accggcacaa gctccgatgt cggcggctac aattacgtgt cctggtatca gcagcacccc 240
ggcaaggccc ctaagctgat gatctacgac gtgtccaaca gaccctccgg cgtgtccaat 300
agattcagcg gcagcaagag cggcaacacc gccagcctga caattagcgg actgcaggcc 360
gaggacgagg ccgattacta ctgtagcagc tacaccagct cctccaccag agtgtttggc 420
accggcacca aagtgaccgt gcttggatcc gaaatcaagc gtacggtggc cgctcccagc 480
gtgttcatct tcccacctag cgacgagcag ctgaagtccg gcacagcctc tgtcgtgtgc 540
ctgctgaaca acttctaccc ccgcgaggcc aaggtgcagt ggaaggtgga caatgccctg 600
cagagcggca acagccagga aagcgtgacc gagcaggaca gcaaggactc cacctacagc 660
ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgc 720
gaagtgaccc accagggcct gtctagcccc gtgaccaaga gcttcaaccg gggcgagtgc 780
taa 783
<210> 155
<211> 2223
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗muCD27-IgG1(N297A)-VH-scFv:Fn14(18D1)
<400> 155
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attccaggtc 120
cagctgcagc agtctggcgc cgaacttgtg aagcctggca gcagcgtgaa gatcagctgt 180
aaagccagcg gctacacctt caccagctac gacatgcact ggatcaagca gcagcccggc 240
aaaggcctgg aatggatcgg ctggatctac cccggcaacg gcaacaccaa gtacaaccag 300
aagttcaacg gcaaggccac actgaccgcc gacatctcta gcagcacagc ctacatgcag 360
ctgagcagcc tgaccagcga agatagcgcc gtgtacttct gcgccaaatg gggctacaac 420
aacttcgact actggggcca gggcgtgatg gtcaccgtgt ctagcagatc cagcagcgcc 480
tctacaaagg gccccagcgt gttccctctg gcccctagca gcaagagcac atctggcgga 540
acagccgccc tgggctgcct cgtgaaggac tactttcccg agcccgtgac cgtgtcctgg 600
aactctggcg ctctgacaag cggcgtgcac acctttccag ccgtgctgca gagcagcggc 660
ctgtactctc tgagcagcgt cgtgacagtg cccagcagct ctctgggcac ccagacctac 720
atctgcaacg tgaaccacaa gcccagcaac accaaggtgg acaagaaggt ggaacccaag 780
agctgcgaca agacccacac ctgtccccct tgtcctgccc ccgaactgct gggaggccct 840
tccgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 900
gtgacctgcg tggtggtgga tgtgtcccac gaggaccctg aagtgaagtt taattggtac 960
gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacgccagc 1020
acctaccggg tggtgtccgt gctgacagtg ctgcaccagg actggctgaa cggcaaagag 1080
tacaagtgca aggtgtccaa caaggccctg cctgccccca tcgagaaaac catcagcaag 1140
gccaagggcc agccccgcga accccaggtg tacacactgc ccccaagcag ggacgagctg 1200
accaagaacc aggtgtccct gacctgtctc gtgaaaggct tctaccccag cgatatcgcc 1260
gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1320
gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagtc ccggtggcag 1380
cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1440
aagtccctga gcctgagccc cggcaagctc gaggaggtgc agctggtgga atctggcggc 1500
ggactggtgc agcctggcgg atctctgaga ctgagctgtg ccgccagcgg cttcaccttc 1560
agcaactact ggatgagctg ggtgcgccag gcccctggca aaggactgga atgggtgtcc 1620
ggcatcaacc caggcggcac ctctacctac tacgccgaca gcgtgaaggg ccggttcacc 1680
atcagccggg acaacgccaa gaacaccctg tacctgcaga tgaacagcct gaagtccgag 1740
gacaccgccg tgtactactg cgccaagcac ctgggcaact ggggcgagta caattactgg 1800
ggccagggca cacaagtgac cgtgtccagt agatctagca ccaagggccc caagctggaa 1860
gagggcgagt tcagcgaggc ccaattgcag agcgccctga cccagcctcc aagcgtgtca 1920
ggctctcctg gcaagaccgt gaccatcagc tgtgctggca ccggcggaga tgtgggctac 1980
agaaacagcg tgtcctggta tcagcagctg cccggcatgg cccccaaact gctgatctac 2040
gacgtggaca agcgggcctc tggcatcacc gacagattca gcggcagcaa gagcggcgat 2100
accgccagcc tgacaatcag cggagtgcag agcgaggacg aggccgacta ctactgtgcc 2160
agccagagaa gcggaatcgc cgccgtgttt ggcggaggca cacacctgac agtgctgggc 2220
taa 2223
<210> 156
<211> 774
<212> DNA
<213> Artificial Sequence
<220>
<223> 抗muCD27-VL
<400> 156
atgaacttcg gcttcagcct gatcttcctg gtgctggtgc tgaagggcgt gcagtgcgaa 60
gtgaagctgg tgccccggca attggactac aaggacgacg acgacaaaga attggacatc 120
cagatgacac agagccctgc cagcctgtct gcctctctgg gagagacagt gtccatcgat 180
tgtctggcca gcgagggcat cagcaacgac ctggcttggt atcagcagaa gtccggcaag 240
agccctcagc tgctgatcaa cagcgccagc agactggaag atggcgtgcc cagcagattt 300
tctggctctg gcagcggcac ccggtacagc ctgaagattt ctggcatgca gcccgaggac 360
gaggccgaat acttctgcct gcaaagctac agaagcccct ggacctttgg cggaggcaca 420
aagctggaac tgaagggatc cgaaatcaag cgtacggtgg ccgctcccag cgtgttcatc 480
ttcccaccta gcgacgagca gctgaagtcc ggcacagcct ctgtcgtgtg cctgctgaac 540
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaatgccct gcagagcggc 600
aacagccagg aaagcgtgac cgagcaggac agcaaggact ccacctacag cctgagcagc 660
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaagtgacc 720
caccagggcc tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaa 774
<210> 157
<211> 750
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 146所示的蛋白质构建体
<400> 157
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Glu Ser Gly Pro Gly
35 40 45
Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly
50 55 60
Gly Ser Ile Ser Ser Gly Asp Tyr Phe Trp Ser Trp Ile Arg Gln Leu
65 70 75 80
Pro Gly Lys Gly Leu Glu Trp Ile Gly His Ile His Asn Ser Gly Thr
85 90 95
Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp
100 105 110
Thr Ser Lys Lys Gln Phe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala
115 120 125
Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Gly Gly Asp Tyr Tyr
130 135 140
Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
145 150 155 160
Arg Ser Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
165 170 175
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
180 185 190
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
195 200 205
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
210 215 220
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
225 230 235 240
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
245 250 255
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
260 265 270
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
275 280 285
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
290 295 300
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
305 310 315 320
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
325 330 335
Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val
340 345 350
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
355 360 365
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
370 375 380
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
385 390 395 400
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
405 410 415
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
420 425 430
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
435 440 445
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
450 455 460
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
465 470 475 480
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu
485 490 495
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Met Gln Pro Gly Gly
500 505 510
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser
515 520 525
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
530 535 540
Ser Ser Ile Tyr Ser Asp Ser Ser Tyr Thr Tyr Tyr Ala Asp Ser Val
545 550 555 560
Lys Ser Arg Phe Thr Ile Ser Thr Asp Asn Ala Lys Asn Thr Leu Tyr
565 570 575
Leu Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Val Tyr Tyr Cys
580 585 590
Ala Gly Ser Ser Asp Tyr Glu Gly Ser Phe Ala Ser Trp Gly Gln Gly
595 600 605
Thr Gln Val Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu
610 615 620
Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu Gln Ser Val Val Thr Gln
625 630 635 640
Pro Pro Ser Leu Ser Ala Ser Pro Gly Ser Ser Val Arg Leu Thr Cys
645 650 655
Thr Leu Ser Ser Gly Asn Ser Val Gly Asn Tyr Asp Ile Ser Trp Tyr
660 665 670
Gln Gln Lys Ala Gly Ser Pro Pro Arg Tyr Leu Leu Tyr Tyr Tyr Ser
675 680 685
Asp Ser Val Lys His Gln Gly Ser Gly Val Pro Ser Arg Phe Ser Gly
690 695 700
Ser Ser Asp Ala Ser Ala Asn Ala Gly Leu Leu Leu Ile Ser Gly Leu
705 710 715 720
Gln Pro Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ala Tyr Lys Ser Gly
725 730 735
Ser His Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly
740 745 750
<210> 158
<211> 255
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 147所示的蛋白质构建体
<400> 158
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
35 40 45
Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser
50 55 60
Gln Gly Ile Ser Arg Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly
65 70 75 80
Gln Ala Pro Ser Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly
85 90 95
Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
100 105 110
Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln
115 120 125
Gln Phe Gly Ser Ser Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Gly
130 135 140
Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
145 150 155 160
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
165 170 175
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
180 185 190
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
195 200 205
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
210 215 220
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
225 230 235 240
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
245 250 255
<210> 159
<211> 741
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 148所示的蛋白质构建体
<400> 159
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Val Gln Ser Gly Ala Glu
35 40 45
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Thr Ala Ser Gly
50 55 60
Phe Asn Ile Lys Asp Tyr Tyr Val His Trp Val Lys Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Met Gly Arg Ile Asp Pro Glu Asp Gly Asp Ser
85 90 95
Lys Tyr Ala Pro Lys Phe Gln Gly Lys Ala Thr Met Thr Ala Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Thr Thr Ser Tyr Tyr Val Gly Thr Tyr Gly
130 135 140
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Ser
145 150 155 160
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
165 170 175
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
180 185 190
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
195 200 205
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
210 215 220
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
225 230 235 240
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
245 250 255
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
260 265 270
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
275 280 285
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
290 295 300
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
305 310 315 320
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
325 330 335
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
340 345 350
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
355 360 365
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
370 375 380
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
385 390 395 400
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
405 410 415
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
420 425 430
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
435 440 445
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
450 455 460
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
465 470 475 480
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln Leu
485 490 495
Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu
500 505 510
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ile Met Met Trp
515 520 525
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Tyr
530 535 540
Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val Lys Gly Arg Phe
545 550 555 560
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn
565 570 575
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ile Lys
580 585 590
Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu Val
595 600 605
Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly
610 615 620
Glu Phe Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Ala Ser
625 630 635 640
Val Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr
645 650 655
Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His
660 665 670
Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg Pro
675 680 685
Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala
690 695 700
Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr
705 710 715 720
Cys Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly Thr
725 730 735
Lys Val Thr Val Leu
740
<210> 160
<211> 256
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 149所示的蛋白质构建体
<400> 160
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
35 40 45
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Ser Ala Ser
50 55 60
Ser Ser Val Ser Tyr Met Leu Trp Phe Gln Gln Lys Pro Gly Lys Ala
65 70 75 80
Pro Lys Leu Leu Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro
85 90 95
Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
100 105 110
Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg
115 120 125
Thr Phe Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
130 135 140
Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
145 150 155 160
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
165 170 175
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
180 185 190
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
195 200 205
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
210 215 220
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
225 230 235 240
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
245 250 255
<210> 161
<211> 525
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 150所示的蛋白质构建体
<400> 161
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Val Gln Ser Gly Ala Glu
35 40 45
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Thr Ala Ser Gly
50 55 60
Phe Asn Ile Lys Asp Tyr Tyr Val His Trp Val Lys Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Met Gly Arg Ile Asp Pro Glu Asp Gly Asp Ser
85 90 95
Lys Tyr Ala Pro Lys Phe Gln Gly Lys Ala Thr Met Thr Ala Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Thr Thr Ser Tyr Tyr Val Gly Thr Tyr Gly
130 135 140
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Ser
145 150 155 160
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
165 170 175
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
180 185 190
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
195 200 205
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
210 215 220
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
225 230 235 240
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
245 250 255
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
260 265 270
Pro Ala Leu Glu Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val
275 280 285
Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
290 295 300
Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly
305 310 315 320
Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr
325 330 335
Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
340 345 350
Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
355 360 365
Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp
370 375 380
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser Thr
385 390 395 400
Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu Gln
405 410 415
Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser
420 425 430
Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn
435 440 445
Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met
450 455 460
Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe Ser
465 470 475 480
Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln
485 490 495
Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser Ser
500 505 510
Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu
515 520 525
<210> 162
<211> 742
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 151所示的蛋白质构建体
<400> 162
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
275 280 285
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
290 295 300
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
305 310 315 320
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
325 330 335
Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
340 345 350
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
355 360 365
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
370 375 380
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
385 390 395 400
Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
405 410 415
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
420 425 430
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
435 440 445
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
450 455 460
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
465 470 475 480
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln
485 490 495
Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg
500 505 510
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ile Met Met
515 520 525
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile
530 535 540
Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val Lys Gly Arg
545 550 555 560
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
565 570 575
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ile
580 585 590
Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu
595 600 605
Val Thr Val Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu
610 615 620
Gly Glu Phe Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Ala
625 630 635 640
Ser Val Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly
645 650 655
Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln
660 665 670
His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg
675 680 685
Pro Ser Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr
690 695 700
Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr
705 710 715 720
Tyr Cys Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly
725 730 735
Thr Lys Val Thr Val Leu
740
<210> 163
<211> 526
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 152所示的蛋白质构建体
<400> 163
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly Asn Gly His Thr
85 90 95
Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu Thr Gly Asp Thr
100 105 110
Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr Ala Arg Ala Phe
130 135 140
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
145 150 155 160
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser
165 170 175
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
180 185 190
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
195 200 205
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
210 215 220
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
225 230 235 240
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
245 250 255
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro
260 265 270
Cys Pro Ala Leu Glu Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu
275 280 285
Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
290 295 300
Thr Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys
305 310 315 320
Gly Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe
325 330 335
Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser
340 345 350
Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
355 360 365
Ala Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val
370 375 380
Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser Ser
385 390 395 400
Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Gln Leu
405 410 415
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
420 425 430
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr
435 440 445
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
450 455 460
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe
465 470 475 480
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
485 490 495
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
500 505 510
Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu
515 520 525
<210> 164
<211> 735
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 153所示的蛋白质构建体
<400> 164
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Glu Val Gln Leu Leu Glu Ser Gly Gly Gly
35 40 45
Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
50 55 60
Phe Thr Phe Ser Ser Tyr Ile Met Met Trp Val Arg Gln Ala Pro Gly
65 70 75 80
Lys Gly Leu Glu Trp Val Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr
85 90 95
Phe Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
100 105 110
Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
115 120 125
Thr Ala Val Tyr Tyr Cys Ala Arg Ile Lys Leu Gly Thr Val Thr Thr
130 135 140
Val Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Arg Ser
145 150 155 160
Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys
165 170 175
Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys
180 185 190
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
195 200 205
Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
210 215 220
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr
225 230 235 240
Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val
245 250 255
Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro
260 265 270
Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
275 280 285
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
290 295 300
Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val
305 310 315 320
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
325 330 335
Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln
340 345 350
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly
355 360 365
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
370 375 380
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
385 390 395 400
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
405 410 415
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
420 425 430
Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
435 440 445
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
450 455 460
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
465 470 475 480
Ser Leu Ser Leu Ser Pro Gly Lys Glu Phe Gln Val Gln Leu Gln Gln
485 490 495
Ser Gly Ala Glu Val Ile Lys Pro Gly Ala Ser Val Lys Leu Ser Cys
500 505 510
Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg
515 520 525
Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Gly
530 535 540
Asn Gly His Thr Asn Tyr Asn Glu Lys Phe Lys Ser Arg Ala Thr Leu
545 550 555 560
Thr Gly Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu
565 570 575
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Phe Thr Thr
580 585 590
Ala Arg Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
595 600 605
Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe Ser
610 615 620
Glu Ala Gln Leu Asp Ile Val Met Thr Gln Ser Pro Ala Phe Leu Ser
625 630 635 640
Val Thr Pro Gly Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Thr
645 650 655
Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro
660 665 670
Lys Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser
675 680 685
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser
690 695 700
Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asp Gly His
705 710 715 720
Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
725 730 735
<210> 165
<211> 260
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 154所示的蛋白质构建体
<400> 165
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Gln Ser Ala Leu Thr Gln Pro Ala Ser Val
35 40 45
Ser Gly Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser
50 55 60
Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro
65 70 75 80
Gly Lys Ala Pro Lys Leu Met Ile Tyr Asp Val Ser Asn Arg Pro Ser
85 90 95
Gly Val Ser Asn Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser
100 105 110
Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys
115 120 125
Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val Phe Gly Thr Gly Thr Lys
130 135 140
Val Thr Val Leu Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
145 150 155 160
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
165 170 175
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
180 185 190
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
195 200 205
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
210 215 220
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
225 230 235 240
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
245 250 255
Arg Gly Glu Cys
260
<210> 166
<211> 740
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 155所示的蛋白质构建体
<400> 166
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Phe Gln Val Gln Leu Gln Gln Ser Gly Ala Glu
35 40 45
Leu Val Lys Pro Gly Ser Ser Val Lys Ile Ser Cys Lys Ala Ser Gly
50 55 60
Tyr Thr Phe Thr Ser Tyr Asp Met His Trp Ile Lys Gln Gln Pro Gly
65 70 75 80
Lys Gly Leu Glu Trp Ile Gly Trp Ile Tyr Pro Gly Asn Gly Asn Thr
85 90 95
Lys Tyr Asn Gln Lys Phe Asn Gly Lys Ala Thr Leu Thr Ala Asp Ile
100 105 110
Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp
115 120 125
Ser Ala Val Tyr Phe Cys Ala Lys Trp Gly Tyr Asn Asn Phe Asp Tyr
130 135 140
Trp Gly Gln Gly Val Met Val Thr Val Ser Ser Arg Ser Ser Ser Ala
145 150 155 160
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
165 170 175
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
180 185 190
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
195 200 205
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
210 215 220
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
225 230 235 240
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
245 250 255
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
260 265 270
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
275 280 285
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
290 295 300
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
305 310 315 320
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
325 330 335
Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
340 345 350
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
355 360 365
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
370 375 380
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
385 390 395 400
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
405 410 415
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
420 425 430
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
435 440 445
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
450 455 460
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
465 470 475 480
Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Glu Glu Val Gln Leu Val
485 490 495
Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser
500 505 510
Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr Trp Met Ser Trp Val
515 520 525
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile Asn Pro
530 535 540
Gly Gly Thr Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr
545 550 555 560
Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser
565 570 575
Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys His Leu Gly
580 585 590
Asn Trp Gly Glu Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
595 600 605
Ser Ser Arg Ser Ser Thr Lys Gly Pro Lys Leu Glu Glu Gly Glu Phe
610 615 620
Ser Glu Ala Gln Leu Gln Ser Ala Leu Thr Gln Pro Pro Ser Val Ser
625 630 635 640
Gly Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Ala Gly Thr Gly Gly
645 650 655
Asp Val Gly Tyr Arg Asn Ser Val Ser Trp Tyr Gln Gln Leu Pro Gly
660 665 670
Met Ala Pro Lys Leu Leu Ile Tyr Asp Val Asp Lys Arg Ala Ser Gly
675 680 685
Ile Thr Asp Arg Phe Ser Gly Ser Lys Ser Gly Asp Thr Ala Ser Leu
690 695 700
Thr Ile Ser Gly Val Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala
705 710 715 720
Ser Gln Arg Ser Gly Ile Ala Ala Val Phe Gly Gly Gly Thr His Leu
725 730 735
Thr Val Leu Gly
740
<210> 167
<211> 257
<212> PRT
<213> Artificial Sequence
<220>
<223> SEQ ID NO: 156所示的蛋白质构建体
<400> 167
Met Asn Phe Gly Phe Ser Leu Ile Phe Leu Val Leu Val Leu Lys Gly
1 5 10 15
Val Gln Cys Glu Val Lys Leu Val Pro Arg Gln Leu Asp Tyr Lys Asp
20 25 30
Asp Asp Asp Lys Glu Leu Asp Ile Gln Met Thr Gln Ser Pro Ala Ser
35 40 45
Leu Ser Ala Ser Leu Gly Glu Thr Val Ser Ile Asp Cys Leu Ala Ser
50 55 60
Glu Gly Ile Ser Asn Asp Leu Ala Trp Tyr Gln Gln Lys Ser Gly Lys
65 70 75 80
Ser Pro Gln Leu Leu Ile Asn Ser Ala Ser Arg Leu Glu Asp Gly Val
85 90 95
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Arg Tyr Ser Leu Lys
100 105 110
Ile Ser Gly Met Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Leu Gln
115 120 125
Ser Tyr Arg Ser Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu
130 135 140
Lys Gly Ser Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
145 150 155 160
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
165 170 175
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
180 185 190
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
195 200 205
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
210 215 220
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
225 230 235 240
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
245 250 255
Cys
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