阅读说明：本技术 一种血型抗体复合物、制备方法 (A kind of blood group antibody compound, preparation method ) 是由 陈晔洲 李勇 王红梅 段生宝 丁少华 魏双施 于 2019-07-23 设计创作，主要内容包括：本发明提供一种血型抗体复合物,结构为血型抗体Ⅰ-桥联二抗-血型抗体Ⅱ。本发明还提供一种制备如上所述的血型抗体复合物的方法,包括如下步骤：S1、选择两个来源于同一生物种属的抗体,即血型抗体Ⅰ和血型抗体Ⅱ；S2、根据血型抗体Ⅰ和血型抗体Ⅱ的种属来源,选择能够同时与血型抗体Ⅰ和血型抗体Ⅱ发生特异性结合的桥联二抗；S3、将血型抗体Ⅰ、血型抗体Ⅱ、桥联二抗按克分子当量数1:1:1混合均匀,室温放置15-30分钟,制得结构为血型抗体Ⅰ-桥联二抗-血型抗体Ⅱ的复合物。本发明所提供的血型抗体复合物的制备方法,利用抗体和其特异性抗体之间特异识别性,可以增加血型抗体的特异性或提高血型抗体的亲合性、效价,操作简单,快捷,应用广泛。(The present invention provides a kind of blood group antibody compound, and structure is the anti-blood group antibody II of I-bridging of blood group antibody two.The present invention also provides a kind of methods for preparing blood group antibody compound as described above, include the following steps: S1, selection two antibody from same biological kind, i.e. blood group antibody I and blood group antibody II；S2, the bridging secondary antibody that can be specifically bound simultaneously with blood group antibody I and blood group antibody II according to the Species origin of blood group antibody I and blood group antibody II, selection；S3, blood group antibody I, blood group antibody II, bridging secondary antibody are uniformly mixed by molar eq number 1:1:1, are placed at room temperature for 15-30 minutes, the compound that structure is the anti-blood group antibody II of I-bridging of blood group antibody two is made.The preparation method of blood group antibody compound provided by the present invention can increase the specificity of blood group antibody or improve affinity, the potency of blood group antibody using specific recognition between antibody and its specific antibody, easy to operate, fast, be widely used.)

1. a kind of blood group antibody compound, which is characterized in that the structure of the blood group antibody compound is I-bridging of blood group antibody Two anti-blood group antibodies II；Wherein, the blood group antibody I and the blood group antibody II are from same biological kind.

2. blood group antibody compound according to claim 1, which is characterized in that the biology includes mouse, chicken, rabbit, people, sheep Camel.

3. blood group antibody compound according to claim 1, which is characterized in that the blood group antibody I and blood group antibody II Resist including source of mouse monoclonal antibody, chicken source widow, rabbit source monoclonal antibody, Human monoclonal antibody, alpaca source monoclonal antibody.

4. blood group antibody compound according to claim 1, which is characterized in that the Species origin and blood of the bridging secondary antibody Type antibody I is different with the Species origin of blood group antibody II.

5. blood group antibody compound according to claim 1, which is characterized in that the bridging secondary antibody include anti-mouse it is mostly anti-and Monoclonal antibody, anti-chicken are mostly anti-, anti-rabbit is mostly anti-, anti-human more anti-, anti-alpaca sources are mostly anti-.

6. blood group antibody compound described in -5 any one according to claim 1, which is characterized in that the blood group antibody I is It include anti-A blood group monoclonal antibody, anti-B blood group monoclonal antibody, anti-RhD blood group monoclonal antibody and mostly anti-with the blood group antibody II.

7. a kind of method for preparing the blood group antibody compound as described in any one of claim 1-6, which is characterized in that packet Include following steps:

S1, selection two blood group antibodies from same biological kind, i.e. blood group antibody I and blood group antibody II；

S2, according to the Species origin of blood group antibody I and blood group antibody II, be selected differently from blood group antibody I and blood group antibody II Species origin and the bridging secondary antibody that can be specifically bound simultaneously with blood group antibody I and blood group antibody II；

S3, blood group antibody I, blood group antibody II, bridging secondary antibody are uniformly mixed by molar eq number 1:1:1, are placed at room temperature for 15- 30 minutes, the blood group compound that structure is the anti-blood group antibody II of I-bridging of blood group antibody two is made.

Technical field

The present invention relates to field of biotechnology more particularly to one kind to mention a kind of blood group antibody compound and preparation method.

Background technique

The immune response ability of antibody includes three aspects: antibody titer, Antibody Avidity, antibody specificity." antibody effect Valence " is biology noun, is the residence time with the physical state of antibody and its in vivo, with its with it is antigen reactive it is how many come Indicate a kind of reaction of its immune effect, it is popular to say the maximum dilution multiple for indicating that antibody be detected." antibody affinity Property " refers to the combination degree between antibody molecule and entire antigen." antibody specificity " refers to antibody for the defined epitope of antigen Selective recognition.

Blood group antibody will be applied in clinical Blood grouping work, and potency and affinity are very important index.It wants The potency and affinity of blood group antibody are improved, the method generallyd use is the cell of screening secretion high-titer high affinity antibody Strain, or the mouse ascites that addition hybridoma induces.The cell strain of screening secretion high-titer high affinity antibody is needed using leaching Bar cell hybridoma technique, is repeated continuously fusion, clone and screening process, this method is time-consuming and laborious, and efficiency is very low, still Obtained cell strain is more stable, and antibody bioactive is preferable.And the mouse ascites induced with hybridoma, although its potency is very Height, but since its is complex in composition, the intracorporal natural antibody of mouse may there are cross reactions with people ABO system, are checked with it It will lead to false positive when blood group, furthermore when Application mouse ascites, cost is big, and yield and quality is unstable, be unsuitable for mass production. Further, since anti-D only has source of people at present, the anti-D of source of mouse monoclonal, the sheet of source of people still are not developed with hybridoma technology Matter limits the cell strain for being difficult screening secretion high-titer high affinity antibody, can not also prepare mouse ascites.

When detecting difficult sample, such as positive reverse type is not consistent, and suspects that there are weak A or weak B antigen, Huo Zhejian When detecting new-born baby blood group due on neonatal red blood cell antigenicity it is very weak, be applied alone anti-A or anti-B to be not easy to detect, need to tie Share anti-AB antibody.But anti-AB antibody is more difficult to get, generally by hybridoma technology, by fusion, gram Grand and screening obtains and A type, Type B, the reaction of AB type red blood cell and the nonreactive anti-AB antibody of O-shaped red blood cell.But this method Randomness is larger, time-consuming and laborious, and efficiency is very low.

Summary of the invention

For overcome the deficiencies in the prior art, described it is an object of the present invention to provide a kind of blood group antibody compound The structure of blood group antibody compound is the anti-blood group antibody II of I-bridging of blood group antibody two.When blood group antibody I and blood group antibody II are When different antibody, structure is the compound of the anti-blood group antibody II of I-bridging of blood group antibody two relative to blood group antibody I or blood group For antibody II, not only with the specificity of blood group antibody I, but also the specificity with blood group antibody II, specificity increase.Work as blood group When antibody I and blood group antibody II are identical antibody, structure is the compound phase of the anti-blood group antibody II of I-bridging of blood group antibody two For I/blood group antibody of blood group antibody II, the potency and affinity of complex antibody are higher than I/blood group antibody of blood group antibody Ⅱ。

To achieve the goals above, the invention is realized by the following technical scheme.

A kind of blood group antibody compound provided by the invention, the structure of the blood group antibody compound are I-bridge of blood group antibody The anti-blood group antibody II of union II；Wherein, the blood group antibody I and the blood group antibody II are from same biological kind.

Preferably, the biology includes but is not limited to mouse, chicken, rabbit, people, alpaca biology.

Preferably, the blood group antibody I and blood group antibody II include but is not limited to source of mouse monoclonal antibody, chicken source widow is anti-, rabbit source is single Anti-, Human monoclonal antibody, alpaca source monoclonal antibody.

Preferably, the Species origin of the bridging secondary antibody is different from the Species origin of blood group antibody I and blood group antibody II.

Preferably, the bridging secondary antibody includes but is not limited to that mostly anti-, the anti-chicken of anti-mouse is mostly anti-, anti-rabbit is mostly anti-, anti-human how anti-, anti- Alpaca is mostly anti-.

Preferably, blood group antibody I and blood group antibody II are the different blood group antibodies from same biological kind.

Preferably, blood group antibody I and blood group antibody II are the identical blood group antibody from same biological kind.

Preferably, the blood group antibody I and the blood group antibody II include but is not limited to anti-A blood group monoclonal antibody, anti-B blood group list Anti-, anti-RhD blood group monoclonal antibody and mostly anti-.

Second object of the present invention provides a kind of method for preparing blood group antibody compound as described above, selects source In the different antibodies or same antibody of same biological kind, i.e. blood group antibody I and blood group antibody II, according to blood group antibody I and blood The Species origin (such as source of mouse) of type antibody II determines required secondary antibody (such as sheep anti mouse or rabbit-anti mouse)；By blood group antibody I, blood group Antibody II and secondary antibody are mixed by molar eq number 1:1:1；Room temperature 15-30 minutes, it was I-bridging of blood group antibody two that structure, which is made, The compound of anti-blood group antibody II.

A kind of method preparing blood group antibody compound as described above provided by the present invention, includes the following steps:

S1, selection two blood group antibodies from same biological kind, i.e. blood group antibody I and blood group antibody II；

S2, according to the Species origin of blood group antibody I and blood group antibody II, be selected differently from blood group antibody I and blood group antibody II Species origin and the bridging secondary antibody that can be specifically bound simultaneously with blood group antibody I and blood group antibody II；

S3, blood group antibody I, blood group antibody II, bridging secondary antibody are uniformly mixed by equivalents 1:1:1, are placed at room temperature for 15-30 Minute, it is compound that the blood group antibody that structure as described in claim 1 is the anti-blood group antibody II of I-bridging of blood group antibody two is made Object.

Compared with prior art, the beneficial effects of the present invention are: it is small with traditional screening high quality cell strain or addition The methods of mouse ascites fluid is compared, and the principle of blood group antibody compound preparation method of the invention is antigen-antibody reaction, utilizes antibody It is prepared between its specific antibody with specific recognition, easy to operate, at low cost, bioactivity is good.Work as the present invention In blood group antibody I and blood group antibody II when being different antibodies, obtained bispecific antibody can be different for two Antigen binding site；When in the present invention blood group antibody I and blood group antibody II be same antibody when, obtained antibody have two A blood group antibody I/II, to reach the potency and affinity for improving blood group antibody I/II.

The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention, And can be implemented in accordance with the contents of the specification, it is described in detail below with some embodiments.A specific embodiment of the invention It is shown in detail by following embodiment.

Detailed description of the invention

The drawings described herein are used to provide a further understanding of the present invention, constitutes part of this application, this hair Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention.In the accompanying drawings:

Fig. 1 is the process flow chart of preparation method of the present invention；

Fig. 2 is the structural schematic diagram of blood group antibody compound of the present invention；

Fig. 3 is the blood group that structure obtained by the present invention is the anti-anti- B blood group monoclonal antibody of A blood group monoclonal antibody-how anti-mouse of sheep anti mouse of mouse The characterization effect picture of antibody complex；

Fig. 4 is the potency comparison diagram of control group and experimental group in one embodiment of the invention；

Fig. 5 is the potency comparison diagram of control group and experimental group in another embodiment of the present invention；

Wherein: 1, blood group antibody I；2, blood group antibody II；3, bridging secondary antibody

Specific embodiment

In order to which invention described herein can be more fully understood by, following embodiment is illustrated.It should understand that these embodiments It being merely to illustrate property purpose and is understood not to limit the invention in any way.

This paper or less is belonged to and is explained.

" molar eq " described herein refers to the molecular amounts of antibody.

" bridging secondary antibody " described herein, also referred to as immunoglobulin, specificity are anti-for blood group antibody I and blood group The kind antibody that body II generates, also referred to as xenoantibody." bridging secondary antibody " is II isotype antigen of blood group antibody I and blood group antibody The antibody of epitope can be specifically bound with blood group antibody I and blood group antibody II simultaneously.