Mesothelin binding proteins

文档序号：496571 发布日期：2022-01-07 浏览：19次中文

阅读说明：本技术 间皮素结合蛋白质 (Mesothelin binding proteins ) 是由霍尔格·韦舍布莱恩·D·莱蒙理查德·J·奥斯汀罗伯特·B·杜布里奇于 2018-05-11 设计创作，主要内容包括：本文公开了MSLN结合蛋白质,其具有改善的结合亲和力和改善的介导表达间皮素的癌细胞的T细胞依赖性杀伤的能力。进一步提供了包含本文公开的结合蛋白质的药物组合物和使用这类制剂的方法。(Disclosed herein are MSLN binding proteins with improved binding affinity and improved ability to mediate T-cell dependent killing of mesothelin-expressing cancer cells. Further provided are pharmaceutical compositions comprising the binding proteins disclosed herein and methods of using such formulations.)

1. A single domain mesothelin binding protein wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 41, 42, 43 or 44 or comprising one or more amino acid residue substitutions with respect to SEQ ID NO 41, 42, 43 or 44.

2. The single domain mesothelin binding protein of claim 1, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO 41 or comprises one or more amino acid residue substitutions relative to SEQ ID NO 41.

3. The single domain mesothelin binding protein according to claim 1 or 2, wherein said protein comprises a sequence comprising a conserved region comprising a sequence identical to SEQ ID NO:42 or comprising one or more amino acid residue substitutions relative to SEQ ID NO: 42.

4. The single domain mesothelin binding protein of claim 1,2 or 3, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID No. 43 or comprises one or more amino acid residue substitutions relative to SEQ ID No. 43.

5. The single domain mesothelin binding protein of claim 1,2 or 3, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:44 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 44.

6. The single domain mesothelin binding protein of claim 1, wherein said protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO: 41; (ii) a stretch of amino acids corresponding to SEQ ID NO 42; (iii) a stretch of amino acids corresponding to SEQ ID NO 43; and (iv) a stretch of amino acids corresponding to SEQ ID NO: 44.

7. A single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

8. A single domain mesothelin binding protein, wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 45, 46, 47, 48, 49 or 50 or comprising one or more amino acid residue substitutions with respect to SEQ ID NO 45, 46, 47, 48, 49 or 50.

9. A single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

10. A single domain mesothelin binding protein, wherein said protein comprises one or more CDRs selected from the group consisting of SEQ ID Nos. 51-56 and 63-179.

Background

The present disclosure provides Mesothelin (MSLN) binding proteins that are useful for diagnosing and treating indications associated with the expression of MSLN. Mesothelin (MSLN) is a GPI-linked membrane-bound tumor antigen MSLN that is overexpressed in ovarian, pancreatic, lung, and triple-negative breast cancers as well as mesotheliomas. Normal tissue expression of MSLN is restricted to the single intercellular cortex lining the pleural, pericardial and peritoneal cavities. Overexpression of MSLN was associated with poor prognosis in lung adenocarcinoma and triple negative breast cancer. MSLN has been used as a cancer antigen for a number of modalities, including immunotoxins, vaccines, antibody drug conjugates, and CAR-T cells. Early signs of clinical efficacy have confirmed MSLN as a target, but therapies with improved efficacy are needed to treat MSLN-expressing cancers.

Disclosure of Invention

One embodiment provides a single domain mesothelin binding protein wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 41, 42, 43 or 44 or comprising one or more amino acid residue substitutions relative to SEQ ID NO 41, 42, 43 or 44. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID NO:41 or comprising one or more amino acid residue substitutions relative to SEQ ID NO: 41. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID NO:42 or comprising one or more amino acid residue substitutions relative to SEQ ID NO: 42. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 43 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 43. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 44 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 44. In some embodiments, the protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO: 41; (ii) a stretch of amino acids corresponding to SEQ ID NO 42; (iii) a stretch of amino acids corresponding to SEQ ID NO 43; and (iv) a stretch of amino acids corresponding to SEQ ID NO: 44.

One embodiment provides a single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

wherein r1 is the same as or comprises one or more amino acid residue substitutions relative to SEQ ID NO 51; r2 is the same as SEQ ID NO:52 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 52; and r3 is the same as SEQ ID NO 53 or comprises one or more amino acid residue substitutions relative to SEQ ID NO 53; and wherein f1, f2, f3 and f4 are framework residues. In some embodiments, the protein comprises a sequence at least 80% identical to a sequence selected from SEQ ID NOs 1-29, 30-40, 58, and 60-62. In some embodiments, the protein comprises one or more modifications that result in humanization of the binding protein. In some embodiments, the modification comprises a substitution, addition, or deletion of an amino acid residue. In some embodiments, the protein comprises from 111 amino acids to 124 amino acids. In some embodiments, the protein comprises a VHH domain derived from a non-human source. In some embodiments, the protein comprises a llama VHH domain. In some embodiments, the epitope is in region I comprising amino acid residues 296-390 of SEQ ID NO:57, region II comprising amino acid residues 391-486 of SEQ ID NO:57 or region III comprising amino acid residues 487-598 of SEQ ID NO: 57.

Some embodiments provide a single domain mesothelin binding protein wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 45, 46, 47, 48, 49 or 50 or comprising one or more amino acid residue substitutions with respect to SEQ ID NO 45, 46, 47, 48, 49 or 50. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID NO:45 or comprising one or more amino acid residue substitutions relative to SEQ ID NO: 45. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 46 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 46. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 47 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 47. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 48 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 48. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID No. 49 or comprising one or more amino acid residue substitutions relative to SEQ ID No. 49. In some embodiments, the protein comprises a conserved region comprising a sequence identical to SEQ ID NO:50 or comprising one or more amino acid residue substitutions relative to SEQ ID NO: 50. In some embodiments, the protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO: 45; (ii) a stretch of amino acids corresponding to SEQ ID NO 46; (iii) a stretch of amino acids corresponding to SEQ ID NO. 47; (iv) a stretch of amino acids corresponding to SEQ ID NO 48; (v) a stretch of amino acids corresponding to SEQ ID NO. 49; and (vi) a stretch of amino acids corresponding to SEQ ID NO: 50.

One embodiment provides a single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

wherein r1 is the same as SEQ ID NO. 54 or comprises one or more amino acid residue substitutions relative to SEQ ID NO. 54; r2 is the same as or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 55; and r3 is the same as or comprises one or more amino acid residue substitutions relative to SEQ ID NO 56; and wherein f1, f2, f3 and f4 are framework residues. In some embodiments, the protein comprises a sequence at least 80% identical to a sequence selected from SEQ ID Nos 30-40, 58, and 60-62. In some embodiments, the protein comprises from 111 amino acids to 119 amino acids. In some embodiments, the protein comprises a VHH domain derived from a non-human source. In some embodiments, the protein comprises a llama VHH domain. In some embodiments, the protein binds to a human mesothelin protein comprising the sequence shown in SEQ ID NO: 57. In some embodiments, the protein binds to an epitope of mesothelin, wherein the epitope is in region I comprising amino acid residues 296-390 of SEQ ID NO:57, region II comprising amino acid residues 391-486 of SEQ ID NO:57 or region III comprising amino acid residues 487-598 of SEQ ID NO: 57. In some embodiments, the binding protein is a chimeric antibody or a humanized antibody. In some embodiments, the binding protein is a single domain antibody. In some embodiments, the binding protein is a humanized single domain antibody.

One embodiment provides a single domain mesothelin binding protein wherein said protein comprises one or more CDRs selected from the group consisting of SEQ ID Nos 51-56 and 63-179. In some embodiments, the protein comprises a CDR1, the CDR1 comprising the sequence set forth in any one of SEQ ID nos. 51, 54, and 63-101. In some embodiments, the protein comprises CDR2, the CDR2 comprising the sequence set forth in any one of SEQ ID Nos. 52, 55 and 102-140. In some embodiments, the protein comprises CDR3, the CDR3 comprising the sequence set forth in any one of SEQ ID Nos. 53, 56 and 141 and 179. In some embodiments, the protein comprises framework region 1(f1), wherein framework region 1 comprises the sequence set forth in any one of SEQ ID Nos. 180-218. In some embodiments, the protein comprises framework region 2(f2), the framework region 2 comprising the sequence set forth in any one of SEQ ID Nos. 219 and 257. In some embodiments, the protein comprises framework region 3(f3), wherein the framework region 3 comprises the sequence set forth in any one of SEQ ID Nos: 258-296. In some embodiments, the protein comprises framework region 4(f4), wherein framework region 4 comprises the sequence set forth in any one of SEQ ID Nos: 297-335. In some embodiments, the protein comprises an amino acid sequence as set forth in any one of SEQ ID nos. 1-40 and 58.

One embodiment provides a polynucleotide encoding a single domain mesothelin binding protein according to any one of the above embodiments. Another embodiment provides a vector comprising the polynucleotide of the above embodiment. Another embodiment provides a host cell transformed with a vector according to the above embodiment.

An embodiment provides a pharmaceutical composition comprising (i) a single domain mesothelin-binding protein according to any one of the above embodiments, a polynucleotide according to any one of the above embodiments, a vector according to any one of the above embodiments or a host cell according to any one of the above embodiments, and (ii) a pharmaceutically acceptable carrier.

Another embodiment provides a method of producing a single domain mesothelin-binding protein according to any one of the above embodiments, said method comprising culturing a host transformed or transfected with a vector comprising a nucleic acid sequence encoding a single domain mesothelin-binding protein according to any one of the above embodiments under conditions permitting expression of the mesothelin-binding protein and recovery and purification of the produced protein from the culture.

One embodiment provides a method of treating or ameliorating a proliferative disease or a neoplastic disease comprising administering to a subject in need thereof a mesothelin binding protein according to any one of the above embodiments. In some embodiments, the subject is a human. In some embodiments, the method further comprises administering an agent in combination with a single domain mesothelin binding protein according to any of the above embodiments. In some embodiments, the single domain mesothelin-binding protein selectively binds to mesothelin-expressing tumor cells. In some embodiments, the single domain mesothelin-binding protein mediates killing of mesothelin-expressing tumor cells by T cells. In some embodiments, the neoplastic disease comprises a solid tumor disease. In some embodiments, the solid tumor disease comprises mesothelioma, lung cancer, gastric cancer, ovarian cancer, or triple negative breast cancer. In some embodiments, the solid tumor disease is metastatic.

Drawings

The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

figure 1 shows the efficacy of exemplary MSLN-targeting trispecific molecules (2a2 and 2a4) containing anti-MSLN binding proteins according to the present disclosure in killing OVCAR8 cells expressing the target protein MSLN.

FIG. 2 shows that a tri-specific MSLN-targeting antigen-binding protein containing an exemplary MSLN binding domain of the disclosure (MH6T) directed T cells from five donors (donor 02; donor 86; donor 41; donor 81; and donor 35) to kill Caov3 cells. The figure also shows that the control trispecific protein (GFP TriTAC) was unable to direct T cells from these five donors (donor 02; donor 86; donor 41; donor 81; and donor 35) to kill Caov3 cells.

Figure 3 shows that a tri-specific MSLN-targeted antigen binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) directed T cells from five donors (donor 02; donor 86; donor 41; donor 81; and donor 35) to kill OVCAR3 cells. The figure also shows that the control trispecific protein (GFP TriTAC) was unable to direct T cells from these five donors (donor 02; donor 86; donor 41; donor 81; and donor 35) to kill OVCAR3 cells.

Figure 4 shows that a tri-specific MSLN-targeted antigen-binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) is able to direct T cells from healthy donors to kill MSLN-expressing cells (OVCAR3 cells; Caov4 cells; OVCAR3 cells; and OVCAR8 cells). This figure also shows that the tri-specific MSLN-targeted antigen-binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) failed to direct T cells from healthy donors to kill MSLN-non-expressing cells (MDAPCa2b cells; and NCI-H510A cells).

FIG. 5 shows that a tri-specific MSLN-targeting antigen-binding protein containing an exemplary MSLN binding domain of the disclosure (MH6T) is able to direct T-cells from cynomolgus monkeys to kill human ovarian cancer cell lines (OVCAR3 cells; Caov3 cells). The figure also shows that the control trispecific protein (GFP TriTAC) is unable to direct T cells from cynomolgus monkeys to kill human ovarian cancer cell lines (OVCAR3 cells; Caov3 cells).

Figure 6 shows that a tri-specific MSLN-targeted antigen-binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) is capable of directing T cells to kill NCI-H2052 mesothelioma cells expressing MSLN in the presence or absence of Human Serum Albumin (HSA).

FIG. 7 shows that a trispecific MSLN-targeted antigen binding protein containing an exemplary MSLN binding domain of the disclosure (MH6T) is capable of activating T cells from four healthy donors (donor 2; donor 86; donor 35; and donor 81) in the presence of MSLN-expressing Caov4 cells, as demonstrated by the secretion of TNF- α by the T cells.

Figure 8 shows that a trispecific MSLN-targeted antigen-binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) was able to activate T cells from four healthy donors (donor 2; donor 86; donor 35; and donor 81) in the presence of MSLN-expressing OVCAR8 cells, as demonstrated by activation of CD69 expression on T cells.

Fig. 9A and 9B show binding of a trispecific MSLN-targeted antigen binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) to a cell line expressing MSLN or a cell line not expressing MSLN. FIG. 9A shows binding to MSLN-expressing cells (Caov3 cells-top left panel; Caov4 cells-top right panel; OVCAR3 cells-bottom left panel; OVCAR8 cells-bottom right panel) that bind to a trispecific MSLN-targeting antigen binding protein containing an exemplary MSLN binding domain of the present disclosure (MH 6T); figure 9A further shows that the control trispecific protein (GFP TriTAC) was unable to bind to the same cell line. Figure 9B shows that neither the trispecific MSLN-targeted antigen-binding protein containing the exemplary MSLN binding domain of the present disclosure (MH6T) nor GFP TriTAC could bind to a cell line that does not express MSLN (MDCA2B cells-left panel; NCI-H510A cells-right panel).

Figure 10 shows binding of a trispecific MSLN-targeted antigen binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) to T cells from four healthy donors (donor 2-top left panel; donor 35-top right panel; donor 41-bottom left panel; donor 81-bottom right panel).

Figure 11 shows that a tri-specific MSLN-targeted antigen-binding protein containing an exemplary MSLN binding domain of the present disclosure (MH6T) is capable of inhibiting tumor growth in NCG mice engrafted with NCI-H292 cells expressing MSLN.

Fig. 12 shows the pharmacokinetic profile of a trispecific MSLN-targeted antigen binding protein containing an exemplary MSLN binding domain of the present disclosure (MH 6T). Serum levels of trispecific MSLN-targeted antigen-binding proteins containing an exemplary MSLN binding domain of the disclosure (MH6T) at different time points after injection into two cynomolgus monkeys are shown.

Detailed Description

While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Certain definitions

The terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent that the terms "includes," including, "" has, "or variants thereof are used in either the detailed description and/or the claims, these terms are intended to be inclusive in a manner similar to the term" comprising.

The term "about" or "approximately" means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which error range will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" may mean within 1 or more than 1 standard deviation, depending on the practice of the given value. Where a particular value is described in the application and claims, unless otherwise stated, the term "about" should be considered to mean an acceptable error range for that particular value.

The terms "individual", "patient" or "subject" are used interchangeably. None of these terms require or are limited to situations characterized by supervision (e.g., continuous or intermittent) by a healthcare worker (e.g., a doctor, a registered nurse, a practicing nurse, a physician's assistant, a caregiver, or a attending care worker).

The term "framework" or "FR" residues (or regions) refers to variable domain residues other than the CDR or hypervariable region residues defined herein. A "human consensus framework" is a framework representing the amino acid residues most frequently occurring in the selection of human immunoglobulin VL or VH framework sequences.

As used herein, "variable region" or "variable domain" refers to the following: certain portions of the variable domains differ widely in sequence between antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domain of the antibody. It is concentrated in three segments called Complementarity Determining Regions (CDRs) or hypervariable regions in the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called the Framework (FR). The variable domains of native heavy and light chains each comprise four FR regions, which predominantly adopt a β -sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the β -sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, together with the CDRs from the other chain, contribute to the formation of the antigen binding site of the antibody (see Kabat et al, Sequences of Proteins of Immunological Interest, fifth edition, National Institute of Health, Bethesda, Md. (1991)). Constant domains, while not directly involved in binding an antibody to an antigen, exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity of the antibody. "variable domain residue numbering according to Kabat" or "amino acid position numbering according to Kabat" and variations thereof refer to the numbering system of the heavy or light chain variable domain used for antibody compilation in Kabat et al, Sequences of Proteins of Immunological Interest, fifth edition Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence can contain fewer or additional amino acids corresponding to a shortening of or insertion into the FRs or CDRs of the variable domains. For example, the heavy chain variable domain may comprise a single amino acid insertion (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc. according to Kabat) after heavy chain FR residue 82. The Kabat numbering of residues for a given antibody can be determined by aligning the antibody sequences in the region of homology with "standard" Kabat numbered sequences. This does not imply that the CDRs of the present disclosure necessarily correspond to the Kabat numbering convention.

As used herein, the term "percent (%) amino acid sequence identity" for a sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular sequence, after the sequences are aligned and gaps, if necessary, are introduced to achieve the maximum percent sequence identity, and no conservative substitutions are considered as part of the sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be accomplished in a variety of ways within the skill in the art, for example, using publicly available computer software such as EMBOSS match, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS needlet, EMBOSS LALIGN, BLAST-2, ALIGN, or megalign (dnastar) software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms required to achieve maximum alignment over the full length of the sequences being compared.

As used herein, "elimination half-life" is used in its ordinary sense, as described in The Pharmaceutical Basis of Therapeutics 21-25(Alfred Goodman Gilman, Louis S.Goodman and Alfred Gilman, sixth edition 1980) by Goodman and Gillman. In short, the term is intended to include a quantitative measure of the time course of drug elimination. The elimination of most drugs is exponential (i.e., follows first order kinetics) because the drug concentration is not generally close to that required for saturation of the elimination process. The rate of the exponential process can be determined by its rate constant k or by its half-life t_1/2Expressed, the rate constant k represents the fractional change per unit time, half-life t_1/2Representing the time required for the process to complete 50%. The units of these two constants are time^-1And time. The first order rate constant and half-life of the reaction are simply related (kXt)_1/20.693) and may be interchanged accordingly. Since the first order elimination kinetics indicate a constant fraction of drug lost per unit time, the log of drug concentration versus time plot is linear at all times after the initial distribution phase (i.e., after drug absorption and distribution is complete). The half-life of drug elimination can be accurately determined from such a graph.

As used herein, the term "knotSynthetic affinity "refers to the affinity of a protein described in this disclosure for its binding target, and is numerically represented using a" Kd "value. If two or more proteins are shown to have comparable binding affinities for their binding targets, then the Kd values for binding of each protein to its binding target are within + -2-fold of each other. If two or more proteins are shown to have comparable binding affinities for a single binding target, then the Kd values for binding of each protein to the single binding target are within + -2-fold of each other. If a protein is shown to bind two or more targets with comparable binding affinity, the Kd values for the binding of the protein to the two or more targets are within + -2-fold of each other. Generally, higher Kd values correspond to weaker binding. In some embodiments, BIAcore is used^TM-2000 or BIAcore^TM-3000(BIAcore, inc., Piscataway, n.j.) to measure "Kd" by radiolabeled antigen binding assay (RIA) or surface plasmon resonance assay. In certain embodiments, BIAcore is also used^TM-2000 or BIAcore^TM-3000(BIAcore, inc., Piscataway, n.j.) the "association rate" or "rate of association" or "association rate" or "kon" and the "dissociation rate" or "rate of dissociation" or "dissociation rate" or "koff" were determined by surface plasmon resonance techniques. In a further embodiment, use is made ofSystems (pall Life sciences) to determine "Kd", "kon" and "koff". In useIn an exemplary method of measuring binding affinity by Systems, a ligand (e.g., biotinylated human or cynomolgus PSMA) is immobilizedStreptavidin capillary sensor tips were surface activated using approximately 20-50 μ g/ml human or cynomolgus monkey MSLN protein following the manufacturer's instructions. Also introduced is PBS/caseinProtein solution is used as a sealant. For association kinetics measurements, MSLN binding protein variants are introduced at a concentration of about 10ng/mL to about 100 μ g/mL, about 50ng/mL to about 5 μ g/mL, or about 2ng/mL to about 20 μ g/mL. In some embodiments, the MSLN binding single domain protein is used at a concentration of about 2ng/mL to about 20 μ g/mL. Complete dissociation was observed in the case of the negative control, i.e. assay buffer without bound protein. The kinetic parameters of the binding reaction are then determined using suitable tools, such as ForteBio software.

Described herein are MSLN binding proteins, pharmaceutical compositions, and nucleic acids, recombinant expression vectors, and host cells for making such MSLN binding proteins. Also provided are methods of preventing and/or treating diseases, conditions, and disorders using the disclosed MSLN binding proteins. The MSLN binding protein is capable of specifically binding to MSLN. In some embodiments, the MSLN binding protein comprises additional domains, such as a CD3 binding domain and an albumin binding domain. Mesothelin (MSLN) and its use in neoplastic diseases

This document relates to mesothelin binding proteins. Mesothelin is a glycoprotein present on the mesothelial lining cell surface of the peritoneal, pleural and pericardial cavities. The mesothelin gene (MSLN) encodes a 71 kilodalton (kDa) precursor protein that is processed into a 40-kDa protein called mesothelin, a glycophosphatidylinositol-anchored glycoprotein present on the cell surface (Chang et al, Proc Natl Acad Sci USA (1996)93: 136-40). Mesothelin cDNA was cloned from a library prepared from the HPC-Y5 cell line (Kojima et al (1995) J.biol.chem.270: 21984-21990). The cDNA was also cloned using the monoclonal antibody K1 which recognizes mesothelioma (Chang and Pastan (1996) Proc. Natl. Acad. Sci. USA 93: 136-40). Mesothelin is a differentiation antigen whose expression in normal human tissues is limited to mesothelial cells lining body cavities such as the pleura, pericardium and peritoneum. Mesothelin is also highly expressed in several different human cancers, including mesothelioma, pancreatic adenocarcinoma, ovarian cancer, gastric and lung adenocarcinoma. (Hassan et al, Eur J Cancer (2008)44:46-53) (Ordonez, Am J Surg Pathol (2003)27: 1418-28; Ho et al, Clin Cancer Res (2007)13: 1571-5). Mesothelin is overexpressed in the vast majority of primary pancreatic adenocarcinomas, with rare and weak expression seen in benign pancreatic tissue. Argani P et al, Clin Cancer Res.2001; 7(12):3862-3868. Epithelial Malignant Pleural Mesothelioma (MPM) ubiquitously expresses mesothelin, whereas sarcoma-like MPM may not. Most serous epithelial ovarian cancers and related primary peritoneal cancers express mesothelin.

Mesothelin also shed from tumor cells as a soluble form of the protein compared to the native membrane-bound form (Hellstrom et al, Cancer epidemibiol biolarkers Prev (2006)15: 1014-20; Ho et al, Cancer epidemi biolarkers Prev (2006)15: 1751). Structurally, mesothelin is expressed on the cell surface as a 60kDa precursor polypeptide that is proteolytically processed into a 31kDa shed component (corresponding to MPF) and a 40kDa membrane bound component (Hassan et al (2004) Clin. cancer. Res.10: 3937-3942). Mesothelin has been shown to interact with CA125 (also known as MUC-16), a mucin-like glycoprotein present on the surface of tumor cells and has previously been identified as an ovarian cancer antigen. In addition, binding of CA125 to membrane-bound mesothelin mediates heterotypic cell adhesion, and CA125 and mesothelin are co-expressed in advanced ovarian adenocarcinoma (Rump, A. et al (2004) J.biol.chem.279: 9190-9198). The expression of mesothelin in the peritoneal lining is associated with a preferred site of metastatic development of ovarian cancer, and mesothelin-CA 125 binding is thought to promote peritoneal metastasis of ovarian tumors (Gubbels, j.a. et al (2006) mol. cancer.5: 50).

Mesothelin is a target of the innate immune response in ovarian cancer and has been proposed as a target for cancer immunotherapy. Bracci L et al Clin Cancer Res.2007; 13(2Pt 1): 644-653; moschella F et al Cancer res.2011; 71(10) 3528-3539; gross G et al FASEB J.1992; 3370-3378; sadelain M et al Nat Rev cancer.2003; 35 to 45 percent (3); muul L M et al blood.2003; 101(7) 2563 and 2569; yee C et al Proc Natl Acad Sci USA.2002; 99(25):16168-16173. The presence of mesothelin-specific CTL in pancreatic cancer patients correlates with overall survival. Thomas A M et al J Exp Med.2004; 200:297-306. In addition, patan and colleagues used soluble antibody fragments of anti-mesothelin antibodies conjugated to immunotoxins to treat cancer patients with mesothelin-positive tumors. This method showed sufficient safety and certain clinical activity in pancreatic cancer. Hassan R et al Cancer Immun.2007; 7:20, and Hassan R et al Clin Cancer Res.2007; 13(17):5144-5149. In ovarian cancer, this treatment strategy produced a minor response according to RECIST criteria and resulted in stable disease in the second patient, whose ascites had also completely resolved.

Mesothelin can also be used as a marker for the diagnosis and prognosis of certain types of cancer, as traces of mesothelin can be detected in the blood of some mesothelin-positive cancer patients (Cristaudo et al, clin. cancer res.13: 5076-. It has been reported that mesothelin can be released into serum by deletion at its carboxy terminus or by proteolytic cleavage from its membrane-bound form (Hassan et al, Clin. cancer Res.10:3937-3942, 2004). An increase in the soluble form of mesothelin can be detected years before malignant mesothelioma develops in asbestos-exposed workers (Creaney and Robinson, Hematol. Oncol. Clin. North am.19: 1025. 1040, 2005). In addition, soluble mesothelin is also elevated in serum from patients with ovarian, pancreatic and lung cancers (Cristaudo et al, Clin. Cancer Res.13:5076-5081, 2007; Hassan et al, Clin. Cancer Res.12:447-453, 2006; Croso et al, Cancer Detect. Prev.30:180-187, 2006). Therefore, mesothelin is a suitable target for disease prevention or treatment methods, and there is a need for effective mesothelin-specific antibodies.

It has been shown that cell surface mature mesothelin comprises three distinct domains, namely region I (comprising residues 296-390), II (comprising residues 391-486) and III (comprising residues 487-598). (Tang et al, A human single-domain antibodies in tissue antigens or activities by targeting an epitope in a spacer close to the cancer cell surface, mol. Can. therapeutics,12(4):416-426, 2013).

The first anti-mesothelin antibodies generated against the therapeutic intervention were designed to interfere with the interaction between mesothelin and CA-125. Phage display identified Fv SS, which was affinity optimized and used to generate recombinant immunotoxin SS1P targeting mesothelin. The MORAB-009 antibody amatuximab of SS1 was also used to recognize a nonlinear epitope in the amino-terminal 64 amino acids within mesothelin region I. Chimeric antigen receptor engineered T cells were also generated using SS1 Fv. Recently, new anti-mesothelin antibodies have been reported that recognize other regions of the mesothelin protein.

There remains a need for additional available options for treating solid tumor diseases associated with mesothelin overexpression, such as ovarian cancer, pancreatic cancer, mesothelioma, lung cancer, gastric cancer, and triple negative breast cancer. In certain embodiments, the present disclosure provides single domain proteins that specifically bind to MSLN on the surface of a tumor target cell.

MSLN binding proteins

In certain embodiments, provided herein are binding proteins, such as anti-MSLN single domain antibodies or antibody variants, that bind to an epitope in the MSLN protein. In some embodiments, the MSLN binding protein binds to a protein comprising the sequence of SEQ ID NO 57. In some embodiments, the MSLN binding protein binds to a protein comprising a truncated sequence compared to SEQ ID NO: 57.

In some embodiments, the MSLN binding proteins disclosed herein recognize full-length mesothelin. In certain instances, the MSLN binding proteins disclosed herein recognize an epitope in mesothelin region I (comprising amino acid residues 296-390 of SEQ ID NO: 57), region II (comprising amino acid residues 391-486 of SEQ ID NO: 57), or region III (comprising amino acid residues 487-598 of SEQ ID NO: 57). It is contemplated that in some embodiments, the MSLN binding proteins of the present disclosure can recognize and bind to epitopes located outside region I, II or III of mesothelin. In still other embodiments, MSLN binding proteins are disclosed that recognize and bind to an epitope different from the moab-009 antibody.

In some embodiments, the MSLN binding proteins of the present disclosure are expressed within multidomain proteins including additional immunoglobulin domains. Such multidomain proteins may act through immunotoxin-based tumor growth inhibition and induction of antibody-dependent cellular cytotoxicity (ADCC). In some embodiments, multidomain proteins comprising an MSLN binding protein of the disclosure exhibit Complement Dependent Cytotoxicity (CDC) activity. In some embodiments, a multidomain protein comprising a MSLN binding protein of the present disclosure exhibits both ADCC and CDC activity on mesothelin-expressing cancer cells.

Furthermore, in some embodiments, where the multidomain protein comprising the MSLN binding protein functions by CDC, the MLSN binding protein can recognize a conformational epitope near the C-terminus of the mesothelin protein on the cell surface. In some embodiments, the mesothelin protein comprises the sequence shown as SEQ ID NO:57 and the C-terminus comprises amino acid residues 539-588.

In some embodiments, the MSLN binding protein is an anti-MSLN antibody or antibody variant. As used herein, the term "antibody variant" refers to variants and derivatives of the antibodies described herein. In certain embodiments, it relates to amino acid sequence variants of the anti-MSLN antibodies described herein. For example, in certain embodiments, amino acid sequence variants of the anti-MSLN antibodies described herein are expected to improve the binding affinity and/or other biological properties of the antibody. Exemplary methods for making amino acid variants include, but are not limited to, introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions from and/or insertions into and/or substitutions of residues within the amino acid sequence of the antibody.

Any combination of deletions, insertions, and substitutions can be made to arrive at the final construct, provided that the final construct possesses the desired properties, e.g., antigen binding. In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include CDRs and framework regions. Examples of such permutations are described below. Amino acid substitutions can be introduced into an antibody of interest and the product screened for a desired activity, e.g., retained/improved antigen binding, reduced immunogenicity, or improved antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). Both conservative and non-conservative amino acid substitutions are contemplated for making antibody variants.

In another example of substitutions used to generate a variant anti-MSLN antibody, one or more hypervariable region residues of the parent antibody are substituted. Typically, the variants are then selected based on an improvement in the desired property compared to the parent antibody, e.g., increased affinity, decreased immunogenicity, increased pH dependence of binding. For example, affinity matured variant antibodies can be generated using, for example, phage display-based affinity maturation techniques, such as those described herein and known in the art.

In some embodiments, the MSLN binding proteins described herein are single domain antibodies, such as heavy chain variable domains (VH) specific for mesothelin, variable domains (VHH) of llama (llama) -derived sdabs, peptides, ligands, or small molecule entities. In some embodiments, the mesothelin-binding domain of the MSLN-binding proteins described herein is any domain that binds to mesothelin, including, but not limited to, domains from monoclonal antibodies, polyclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies. In certain embodiments, the MSLN binding protein is a single domain antibody. In other embodiments, the MSLN binding protein is a peptide. In a further embodiment, the MSLN binding protein is a small molecule.

Generally, it should be noted that the term single domain antibody as used herein in its broadest sense is not limited to a particular biological source or a particular method of preparation. A single domain antibody is an antibody whose complementarity determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies that naturally do not contain a light chain, single domain antibodies derived from conventional 4 chain antibodies, engineered antibodies, and single domain scaffolds other than those derived from antibodies. The single domain antibody may be any single domain antibody in the art, or any future single domain antibody. Single domain antibodies may be derived from any species, including but not limited to mouse, human, camel, llama, goat, rabbit, cow. For example, in some embodiments, a single domain antibody of the present disclosure is obtained by: (1) by isolating the VHH domain of a naturally occurring heavy chain antibody; (2) by expressing a nucleotide sequence encoding a naturally occurring VHH domain; (3) by "humanization" of naturally occurring VHH domains or by expression of nucleic acids encoding such humanized VHH domains; (4) by "camelising" a naturally occurring VH domain from any animal species, particularly from a mammalian species, such as from a human, or by expressing a nucleic acid encoding such a camelised VH domain; (5) by "camelization" of a "domain antibody" or "Dab", or by expression of a nucleic acid encoding such a camelized VH domain; (6) preparing a protein, polypeptide or other amino acid sequence by using synthetic or semi-synthetic techniques; (7) preparing a nucleic acid encoding a single domain antibody by using nucleic acid synthesis techniques known in the art, and then expressing the nucleic acid thus obtained; and/or (8) by any combination of one or more of the foregoing.

In one embodiment, the single domain antibody corresponds to the VHH domain of a naturally occurring heavy chain antibody directed against MSLN. As further described herein, such VHH sequences can generally be generated or obtained by the following methods: suitably immunizing a llama species with MSLN (i.e. to generate an immune response and/or heavy chain antibodies against MSLN), obtaining a suitable biological sample (such as a blood sample, serum sample or B cell sample) from the llama, and generating VHH sequences against MSLN from the sample using any suitable technique known in the art.

In another embodiment, such naturally occurring VHH domains directed against MSLNs are obtained from a naive library of camelidae VHH sequences, such a library being screened using MSLNs or at least one part, fragment, antigenic determinant or epitope thereof, e.g. by using one or more screening techniques known in the art. Such libraries and techniques are described, for example, in WO 99/37681, WO 01/90190, WO 03/025020 and WO 03/035694. Alternatively, improved synthetic or semi-synthetic libraries derived from naive VHH libraries are used, such as VHH libraries obtained from naive VHH libraries by techniques such as random mutagenesis and/or CDR shuffling, for example as described in WO 00/43507.

In a further embodiment, yet another technique for obtaining VHH sequences directed to MSLN involves suitably immunizing a transgenic mammal capable of expressing heavy chain antibodies (i.e., to generate an immune response against MSLN and/or heavy chain antibodies), obtaining a suitable biological sample (such as a blood sample, serum sample, or B cell sample) from the transgenic mammal, and then generating VHH sequences directed to MSLN from the sample using any suitable technique known in the art. For example, rats or mice expressing heavy chain antibodies and other methods and techniques described in WO 02/085945 and WO 04/049794 may be used for this purpose.

In some embodiments, the anti-MSLN antibodies described herein comprise single domain antibodies having an amino acid sequence corresponding to the amino acid sequence of a naturally occurring VHH domain, but which have been "humanized", i.e. by replacing one or more amino acid residues in the amino acid sequence of said naturally occurring VHH sequence (in particular in the framework sequence) with one or more amino acid residues present at corresponding positions in the VH domain of a conventional 4-chain antibody from a human (e.g. as described above). This may be done in a manner known in the art and will be apparent to those skilled in the art, e.g., based on the further description herein. Furthermore, it should be noted that such humanized anti-MSLN single domain antibodies of the present disclosure are obtained in any suitable manner known per se (i.e., as shown in points (1) - (8) above), and are therefore not strictly limited to polypeptides obtained using a polypeptide comprising a naturally occurring VHH domain as a starting material. In some further embodiments, a single domain MSLN antibody as described herein comprises a single domain antibody having an amino acid sequence corresponding to the amino acid sequence of a naturally occurring VH domain, but which has been "camelised", i.e. by replacing one or more amino acid residues in the amino acid sequence of a naturally occurring VH domain from a conventional 4 chain antibody with one or more amino acid residues present at corresponding positions in the VHH domain of the heavy chain antibody. Such "camelised" substitutions are preferably inserted at amino acid positions forming and/or present at the VH-VL interface and/or at so-called camelid tag residues (see e.g. WO 94/04678, and Davies and Riechmann (1994 and 1996)). Preferably, the VH sequences used as starting material or starting point for generating or designing camelised single domains are preferably VH sequences from mammals, more preferably VH sequences from humans, such as VH3 sequences. It should be noted, however, that in certain embodiments, such camelized anti-MSLN single domain antibodies of the present disclosure are obtained in any suitable manner known in the art (i.e., as shown in points (1) - (8) above), and are therefore not strictly limited to polypeptides obtained using a polypeptide comprising a naturally occurring VH domain as a starting material. For example, "humanization" and "camelization" are both performed by providing a nucleotide sequence encoding a naturally occurring VHH domain or VH domain, respectively, and then altering one or more codons in the nucleotide sequence in such a way that the new nucleotide sequence encodes a "humanized" or "camelized" single domain antibody, respectively, as further described herein. The nucleic acid may then be expressed to provide the desired anti-MSLN single domain antibodies of the present disclosure. Alternatively, in other embodiments, the amino acid sequence of a desired humanized or camelized anti-MSLN single domain antibody of the present disclosure is designed based on the amino acid sequence of a naturally occurring VHH domain or VH domain, respectively, and then synthesized de novo using known peptide synthesis techniques. In some embodiments, the nucleotide sequence encoding a desired humanized or camelized anti-MSLN single domain antibody of the present disclosure is designed based on the amino acid sequence or nucleotide sequence of a naturally occurring VHH domain or VH domain, respectively, and then synthesized de novo using known nucleic acid synthesis techniques, followed by expression of the thus obtained nucleic acid using known expression techniques, to provide the desired anti-MSLN single domain antibody of the present disclosure.

Other suitable methods and techniques for obtaining the anti-MSLN single domain antibodies of the present disclosure and/or nucleic acids encoding the same, starting from naturally-occurring VH sequences or VHH sequences, include, for example, combining one or more portions of one or more naturally-occurring VH sequences, such as one or more Framework (FR) sequences and/or Complementarity Determining Region (CDR) sequences, one or more portions of one or more naturally-occurring VHH sequences, such as one or more FR sequences or CDR sequences, and/or one or more synthetic or semi-synthetic sequences in a suitable manner to provide the anti-MSLN single domain antibodies of the present disclosure or nucleotide sequences or nucleic acids encoding the same.

In some embodiments, it is contemplated that the MSLN binding protein is quite small, and in some embodiments no greater than 25kD, no greater than 20kD, no greater than 15kD, or no greater than 10 kD. In certain instances, the MSLN binding protein, if a peptide or small molecule entity, is 5kD or less.

In some embodiments, the MSLN binding protein is an anti-MSLN specific antibody comprising a heavy chain variable complementarity determining region CDR1, a heavy chain variable CDR2, a heavy chain variable CDR3, a light chain variable CDR1, a light chain variable CDR2, and a light chain variable CDR 3. In some embodiments, the MSLN-binding protein comprises any domain that binds to MSLN, including but not limited to domains from: monoclonal antibodies, polyclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, or antigen binding fragments such as single domain antibodies (sdabs), Fab', f (ab)2, and Fv fragments, fragments consisting of one or more CDRs, single chain antibodies (e.g., single chain Fv fragment (scFv)), disulfide stabilized Fv (dsfv) fragments, heteroconjugate antibodies (e.g., bispecific antibodies), pFv fragments, heavy chain monomers or dimers, light chain monomers or dimers, and dimers consisting of one heavy chain and one light chain. In some embodiments, the MSLN binding protein is a single domain antibody. In some embodiments, the anti-MSLN single domain antibody comprises a heavy chain variable Complementarity Determining Region (CDR), CDR1, CDR2, and CDR 3.

In some embodiments, the MSLN binding proteins of the present disclosure are polypeptides comprising an amino acid sequence consisting of four framework regions/sequences (f1-f4) interrupted by three complementarity determining regions/sequences, as shown in the following formula: f1-r1-f2-r2-f3-r3-f4, wherein r1, r2 and r3 are complementarity determining regions CDR1, CDR2 and CDR3, respectively, and f1, f2, f3 and f4 are framework residues. The framework residues of the MSLN binding proteins of the present disclosure comprise, for example, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, or 94 amino acid residues, and the complementarity determining region comprises, for example, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 amino acid residues. In some embodiments, the MSLN binding protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs 1-40.

In some embodiments, the CDR1 comprises the amino acid sequence shown as SEQ ID No. 51 or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 51. In some embodiments, the CDR2 comprises the sequence shown as SEQ ID No. 52, or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 52. In some embodiments, the CDR3 comprises the sequence shown as SEQ ID No. 53, or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 53.

In some embodiments, the CDR1 comprises the amino acid sequence shown as SEQ ID No. 54 or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 54. In some embodiments, the CDR2 comprises the sequence shown as SEQ ID No. 55, or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 55. In some embodiments, the CDR3 comprises the sequence shown as SEQ ID No. 56, or a variant having one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions in SEQ ID No. 56.

In certain examples, the MSLN binding proteins of the present disclosure comprise one or more conserved regions. The conserved region comprises a sequence as set forth in SEQ ID NOs: 41-50, or a variant comprising one or more amino acid residue substitutions relative to the sequence. Exemplary embodiments include MSLN binding proteins comprising one or more conserved regions selected from SEQ ID NOs 41-44, or variants comprising one or more amino acid residue substitutions relative to the sequence. In some cases, the MSLN binding protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO:41, (ii) a stretch of amino acids corresponding to SEQ ID NO:42, (iii) a stretch of amino acids corresponding to SEQ ID NO:43, and (iv) a stretch of amino acids corresponding to SEQ ID NO: 44.

Other exemplary embodiments include MSLN binding proteins comprising one or more conserved regions selected from SEQ ID NOS: 45-50, or variants comprising one or more amino acid residue substitutions relative to the sequence. In some cases, the MSLN binding protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO:45, (ii) a stretch of amino acids corresponding to SEQ ID NO:46, (iii) a stretch of amino acids corresponding to SEQ ID NO:47, (iv) a stretch of amino acids corresponding to SEQ ID NO:48, (v) a stretch of amino acids corresponding to SEQ ID NO:49, and (vi) a stretch of amino acids corresponding to SEQ ID NO: 50.

In various embodiments, an MSLN binding protein of the present disclosure is at least about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to an amino acid sequence selected from SEQ ID NOs 1-29, 58, and 60-62.

In various embodiments, the complementarity determining region of an MSLN binding protein of the present disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID No. 51 or SEQ ID No. 54.

In various embodiments, complementarity determining region 1(CDR1) of an MSLN binding protein of the present disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to an amino acid sequence set forth in any of SEQ ID NOs 63-101.

In various embodiments, complementarity determining region 2(CDR2) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in any one of SEQ ID NO: 102-140.

In various embodiments, the complementarity determining region 3(CDR3) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in any of SEQ ID NO. 141-179.

In various embodiments, framework region 1(f1) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical to the amino acid sequence set forth in any of SEQ ID NO: 180-.

In various embodiments, framework region 1(f1) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in any of SEQ ID NO: 219-257.

In various embodiments, framework region 2(f2) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in any of SEQ ID NO: 258-.

In various embodiments, framework region 3(f3) of an MSLN binding protein of the disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% identical to the amino acid sequence set forth in any of SEQ ID NO: 297-.

In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 1. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 2. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 3. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 4. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 5. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 6. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 7. In some embodiments, the MSLN binding protein according to any of the embodiments above is a single domain antibody comprising the sequence of SEQ ID NO 8. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 9. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 10. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 11. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 12. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 13. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 14. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 15. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 16. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 17. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 18. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 19. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID NO: 20. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 21. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 22. In some embodiments, the MSLN binding protein according to any of the embodiments above is a single domain antibody comprising the sequence of SEQ ID NO 23. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 24. In some embodiments, the MSLN binding protein according to any of the embodiments above is a single domain antibody comprising the sequence of SEQ ID NO. 25. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 26. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 27. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 28. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a single domain antibody comprising the sequence of SEQ ID No. 28.

In some embodiments, the MSLN binding protein according to any of the embodiments above is a humanized single domain antibody comprising the sequence of SEQ ID NO 30. In some embodiments, the MSLN binding protein according to any of the embodiments above is a humanized single domain antibody comprising the sequence of SEQ ID NO 31. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID No. 32. In some embodiments, the MSLN binding protein according to any of the embodiments above is a humanized single domain antibody comprising the sequence of SEQ ID NO 33. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID No. 34. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID No. 35. In some embodiments, the MSLN binding protein according to any of the embodiments above is a humanized single domain antibody comprising the sequence of SEQ ID NO: 36. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID No. 37. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID NO: 38. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID No. 39. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID NO: 40. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID NO: 58. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID NO: 60. In some embodiments, the MSLN binding protein according to any of the embodiments above is a humanized single domain antibody comprising the sequence of SEQ ID NO 61. In some embodiments, the MSLN binding protein according to any one of the above embodiments is a humanized single domain antibody comprising the sequence of SEQ ID NO: 62.

In some embodiments, the MSLN binding protein is cross-reactive to human and cynomolgus monkey mesothelin. In some embodiments, the MSLN binding protein is specific for human mesothelin. In certain embodiments, the MSLN binding proteins disclosed herein bind to human mesothelin with a human kd (hkd). In certain embodiments, the MSLN binding proteins disclosed herein bind to cynomolgus monkey mesothelin with cynomolgus monkey kd (cdd). In certain embodiments, the MSLN binding proteins disclosed herein bind to cynomolgus monkey mesothelin and human mesothelin with cynomolgus monkey kd (cdd) and human kd (hkd), respectively. In some embodiments, the MSLN binding protein binds to human and cynomolgus monkey mesothelin with comparable binding affinities (i.e., no more than a ± 10% difference between the hKd and cKd values). In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 500 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 450 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 400 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 350 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 300 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 250 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 200 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 150 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 100 nM. In some embodiments, the hKd and cKd are in the range of about 0.1nM to about 90 nM. In some embodiments, the hKd and cKd are in the range of about 0.2nM to about 80 nM. In some embodiments, the hKd and cKd are in the range of about 0.3nM to about 70 nM. In some embodiments, the hKd and cKd are in the range of about 0.4nM to about 50 nM. In some embodiments, the hKd and cKd are in the range of about 0.5nM to about 30 nM. In some embodiments, the hKd and cKd are in the range of about 0.6nM to about 10 nM. In some embodiments, the hKd and cKd are in the range of about 0.7nM to about 8 nM. In some embodiments, the hKd and cKd are in the range of about 0.8nM to about 6 nM. In some embodiments, the hKd and cKd are in the range of about 0.9nM to about 4 nM. In some embodiments, the hKd and cKd are in the range of about 1nM to about 2 nM.

In some embodiments, any of the foregoing MSLN binding proteins (e.g., the anti-MSLN single domain antibodies of SEQ ID NOS: 1-40 and 58) are affinity peptides labeled for ease of purification. In some embodiments, the affinity peptide tag is six consecutive histidine residues, also known as 6X-his.

In certain embodiments, MSLN binding proteins according to the present disclosure can be incorporated into MSLN-targeted trispecific proteins. In some examples, the trispecific binding protein comprises a CD3 binding domain, a Human Serum Albumin (HSA) binding domain, and an anti-MSLN binding domain according to the present disclosure. In some cases, the trispecific binding protein comprises the above domains in the following orientations: MSLN-HSA-CD 3.

In certain embodiments, the MSLN-binding proteins of the present disclosure preferentially bind membrane-bound mesothelin relative to soluble mesothelin. Membrane-bound mesothelin refers to the presence of mesothelin in or on the cell membrane surface of mesothelin-expressing cells. Soluble mesothelin refers to mesothelin that is no longer present in or on the cell membrane surface of cells that are expressing or have expressed mesothelin. In certain instances, soluble mesothelin is present in the blood and/or lymphatic circulation of the subject. In one embodiment, the MSLN binding protein binds to membrane-bound mesothelin at least 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 100-fold, 500-fold, or 1000-fold greater than the binding to soluble mesothelin. In one embodiment, the antigen binding proteins of the present disclosure preferentially bind membrane-bound mesothelin, 30-fold greater than binding to soluble mesothelin. Preferential binding of the antigen binding protein to membrane-bound MSLN relative to soluble MSLN can be readily determined using assays well known in the art.

Integration into Chimeric Antigen Receptors (CAR)

In certain examples, MSLN binding proteins of the present disclosure, e.g., anti-MSLN single domain antibodies, can be incorporated into a Chimeric Antigen Receptor (CAR). Engineered immune effector cells, such as T cells or NK cells, can be used to express a CAR that includes an anti-MSLN single domain antibody described herein. In one embodiment, a CAR comprising an anti-MSLN single domain antibody described herein is linked to a transmembrane domain via a hinge region and further linked to a co-stimulatory domain, e.g., a functional signaling domain obtained from OX40, CD27, CD28, CD5, ICAM-1, LFA-1(CD11a/CD18), ICOS (CD278), or 4-1 BB. In some embodiments, the CAR further comprises a sequence encoding an intracellular signaling domain, such as 4-1BB and/or CD3 ζ.

Tumor growth reducing Properties

In certain embodiments, the MSLN binding proteins of the present disclosure reduce the growth of tumor cells in vivo when administered to a subject having mesothelin-expressing tumor cells. The measurement of the reduction of tumor cell growth can be determined by a number of different methods known in the art. Non-limiting examples include direct measurement of tumor size, measurement of excised tumor mass and comparison to control subjects, measurement by imaging techniques (e.g., CT or MRI) that may or may not use isotope or luminescent molecule (e.g., luciferase) enhanced assays, and the like. In particular embodiments, administration of an antigen-binding agent of the present disclosure results in at least about a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in vivo growth of tumor cells as compared to a control antigen-binding agent, with a reduction in tumor growth of about 100% indicating a complete response and tumor disappearance. In further embodiments, administration of an antigen-binding agent of the present disclosure results in a reduction in vivo growth of tumor cells by about 50-100%, about 75-100%, or about 90-100% as compared to a control antigen-binding agent. In further embodiments, administration of an antigen-binding agent of the present disclosure results in a reduction in vivo growth of tumor cells by about 50-60%, about 60-70%, about 70-80%, about 80-90%, or about 90-100% as compared to a control antigen-binding agent.

MSLN binding protein modifications

The MSLN binding proteins described herein include derivatives or analogs in which (i) an amino acid is replaced with an amino acid residue that is not encoded by the genetic code, (ii) the mature polypeptide is fused to another compound such as polyethylene glycol, or (iii) additional amino acids are fused to the protein, such as a leader or secretory sequence or a sequence used to block an immunogenic domain and/or for purification of the protein.

Typical modifications include, but are not limited to, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cystine, formation of pyroglutamate, formylation, gamma carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenization, sulfation, transfer RNA mediated addition of amino acids to proteins, such as arginylation and ubiquitination.

Modifications are made at any position in the MSLN binding proteins described herein, including the peptide backbone, the amino acid side chain, and the amino or carboxy terminus. Some common peptide modifications that can be used to modify MSLN-binding proteins include glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation, blocking of amino or carboxyl groups in the polypeptide or both by covalent modification, and ADP-ribosylation.

Polynucleotides encoding MSLN binding proteins

In some embodiments, polynucleotide molecules encoding the MSLN binding proteins described herein are also provided. In some embodiments, the polynucleotide molecule is provided in the form of a DNA construct. In other embodiments, the polynucleotide molecule is provided in the form of a messenger RNA transcript.

The polynucleotide molecule is constructed by known methods, for example by combining a gene encoding an anti-MSLN binding protein operably linked to a suitable promoter and optionally a suitable transcription terminator and expressing it in bacteria or other suitable expression systems such as CHO cells.

In some embodiments, the polynucleotide is inserted into a vector, preferably an expression vector, which represents a further embodiment. The recombinant vector can be constructed according to known methods. Vectors of particular interest include plasmids, phagemids, phage derivatives, viruses (virii) (e.g., retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, lentiviruses, and the like), and cosmids.

A variety of expression vector/host systems are available to contain and express polynucleotides encoding the MSLN-binding protein polypeptides. Examples of expression vectors are pSKK (Le Gall et al, J Immunol Methods (2004)285(1):111-27) for expression in E.coli, pcDNA5(Invitrogen), PICHIAPINK for expression in mammalian cells^TM Yeast Expression Systems(Invitrogen)、BACUVANCE^TM Baculovirus Expression System(GenScript)。

Thus, in some embodiments, a MSLN binding protein as described herein is produced by: the vector encoding the protein as described above is introduced into a host cell and the host cell is cultured under conditions allowing the expression of the protein domain, which may be isolated and optionally further purified.

Pharmaceutical composition

In some embodiments, also provided are pharmaceutical compositions comprising a MSLN binding protein described herein, a vector comprising a polynucleotide encoding a polypeptide of the MSLN binding protein, or a host cell transformed with the vector, and at least one pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" includes, but is not limited to, any carrier that does not interfere with the effectiveness of the biological activity of the ingredients and is non-toxic to the patient to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions such as oil-in-water emulsions, various types of wetting agents, sterile solutions, and the like. Such carriers can be formulated by conventional methods and can be administered to a subject in a suitable dosage. Preferably, the composition is sterile. These compositions may also contain adjuvants such as preservatives, emulsifiers and dispersants. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents. Another embodiment provides one or more of the above-described binding proteins, such as an anti-MSLN single domain antibody or antigen-binding fragment thereof, packaged in lyophilized form or in an aqueous medium.

In some embodiments of the pharmaceutical composition, the MSLN binding protein described herein is encapsulated in a nanoparticle. In some embodiments, the nanoparticle is a fullerene, a liquid crystal, a liposome, a quantum dot, a superparamagnetic nanoparticle, a dendrimer, or a nanorod. In other embodiments of the pharmaceutical composition, the MSLN binding protein is attached to a liposome. In some cases, the MSLN binding protein is conjugated to the liposome surface. In some cases, the MSLN binding protein is encapsulated within the shell of the liposome. In some cases, the liposome is a cationic liposome.

The MSLN binding proteins described herein are contemplated for use as medicaments. Administration is achieved in different ways, e.g. by intravenous, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration. In some embodiments, the route of administration depends on the type of therapy and the type of compound included in the pharmaceutical composition. The dosage regimen will be determined by the attending physician in light of other clinical factors. The dosage for any one patient will depend upon a number of factors including the patient's size, body surface area, age, sex, the particular compound to be administered, time and route of administration, the type of therapy, general health, and other drugs being administered concurrently. An "effective dose" refers to an amount of active ingredient sufficient to affect the course and severity of the disease, resulting in the alleviation or remission of such pathology, and can be determined using known methods.

In some embodiments, the MSLN conjugates of the present disclosure are administered at a dose of up to 10mg/kg at a frequency of once per week. In some cases, the dose ranges from about 1ng/kg to about 10 mg/kg. In some embodiments, the dose is about 1ng/kg to about 10ng/kg, about 5ng/kg to about 15ng/kg, about 12ng/kg to about 20ng/kg, about 18ng/kg to about 30ng/kg, about 25ng/kg to about 50ng/kg, about 35ng/kg to about 60ng/kg, about 45ng/kg to about 70ng/kg, about 65ng/kg to about 85ng/kg, about 80ng/kg to about 1 μ g/kg, about 0.5 μ g/kg to about 5 μ g/kg, about 2 μ g/kg to about 10 μ g/kg, about 7 μ g/kg to about 15 μ g/kg, about 12 μ g/kg to about 25 μ g/kg, about 20 μ g/kg to about 50 μ g/kg, about 35 μ g/kg to about 70 μ g/kg, About 45 to about 80, about 65 to about 90, about 85 to about 0.1, about 0.095 to about 10 mg/kg. In some cases, the dose is from about 0.1mg/kg to about 0.2mg/kg, from about 0.25mg/kg to about 0.5mg/kg, from about 0.45mg/kg to about 1mg/kg, from about 0.75mg/kg to about 3mg/kg, from about 2.5mg/kg to about 4mg/kg, from about 3.5mg/kg to about 5mg/kg, from about 4.5mg/kg to about 6mg/kg, from about 5.5mg/kg to about 7mg/kg, from about 6.5mg/kg to about 8mg/kg, from about 7.5mg/kg to about 9mg/kg, or from about 8.5mg/kg to about 10 mg/kg. In some embodiments, the frequency of administration is about less than once per day, once every other day, less than once per day, twice per week, once per 7 days, once per two weeks, once per three weeks, once per four weeks, or once per month. In some cases, the frequency of administration is once per week. In some cases, the frequency of administration is once per week at a dose of up to 10 mg/kg. In some cases, the duration of administration is from about 1 day to about 4 weeks or more

Method of treatment

In some embodiments, also provided herein are methods and uses for stimulating the immune system of an individual in need thereof comprising administering an MSLN binding protein described herein. In some cases, administration of the MSLN binding proteins described herein induces and/or maintains cytotoxicity to cells expressing the target antigen. In some cases, the cell expressing the target antigen is a cancer or tumor cell, a virus-infected cell, a bacteria-infected cell, an autoreactive T or B cell, a damaged red blood cell, an arterial plaque, or a fibrotic tissue.

Also provided herein are methods and uses for treating a disease, disorder or condition associated with a target antigen, comprising administering to an individual in need thereof an MSLN binding protein described herein or a multispecific binding protein comprising the MSLN binding protein. Diseases, disorders or conditions associated with the target antigen include, but are not limited to, viral infections, bacterial infections, autoimmune diseases, transplant rejection, atherosclerosis or fibrosis. In other embodiments, the disease, disorder or condition associated with the target antigen is a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergy, a parasitic reaction, a graft-versus-host disease or a host-versus-graft disease. In one embodiment, the disease, disorder or condition associated with the target antigen is cancer. Cancers that may be treated, prevented or controlled with the MSLN binding proteins of the present disclosure and methods of use thereof include, but are not limited to, cancers of epithelial cell origin. Examples of such cancers include the following: leukemias, such as, but not limited to, acute leukemia, acute lymphocytic leukemia, acute myelogenous leukemia, such as myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia, and myelodysplastic syndrome; chronic leukemias, such as, but not limited to, chronic myelogenous (myelogenous) leukemia, chronic lymphocytic leukemia, hairy cell leukemia; polycythemia vera; lymphomas such as, but not limited to, hodgkin's disease, non-hodgkin's disease; multiple myeloma, such as, but not limited to, smoldering multiple myeloma, non-secretory myeloma, osteosclerotic myeloma, plasma cell leukemia, solitary plasmacytoma, and extramedullary plasmacytoma; waldenstrom's macroglobulinemia; monoclonal gammopathy of unknown significance; benign monoclonal gammopathy; heavy chain disease; bone and connective tissue sarcomas such as, but not limited to, sarcomas of the bone, osteosarcoma, chondrosarcoma, ewing's sarcoma, malignant giant cell tumor, bone fibrosarcoma, chordoma, periosteal sarcoma, soft tissue sarcoma, angiosarcoma, fibrosarcoma, kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, schwannoma, rhabdomyosarcoma, synovial sarcoma; brain tumors such as, but not limited to, glioma, astrocytoma, brain stem glioma, ependymoma, oligodendroglioma, non-glioma, acoustic schwannoma, craniopharyngioma, medulloblastoma, meningioma, pinealocytoma, pinealoblastoma, primary brain lymphoma; breast cancers including, but not limited to, ductal, adenocarcinoma, lobular (small cell) carcinoma, intraductal, medullary, mucinous, tubular, papillary, paget's disease, and inflammatory breast cancers; adrenal cancer such as, but not limited to, pheochromocytoma and adrenocortical carcinoma; thyroid cancer such as, but not limited to, papillary or follicular thyroid cancer, medullary thyroid cancer, and anaplastic thyroid cancer; pancreatic cancers such as, but not limited to, insulinoma, gastrinoma, glucagonoma, viral tumors, somatostatin-secreting tumors, and carcinoid or islet cell tumors; pituitary cancers such as, but not limited to, cushing's disease, prolactin-secreting tumors, acromegaly, and diabetes insipidus; eye cancers such as, but not limited to, ocular melanoma, such as iris melanoma, choroidal melanoma, and ciliary melanoma, and retinoblastoma; vaginal cancers such as squamous cell carcinoma, adenocarcinoma, and melanoma; vulvar cancers, such as squamous cell carcinoma, melanoma, adenocarcinoma, basal cell carcinoma, sarcoma, and paget's disease; cervical cancers such as, but not limited to, squamous cell carcinoma and adenocarcinoma; uterine cancers such as, but not limited to, endometrial carcinoma and uterine sarcoma; ovarian cancers such as, but not limited to, ovarian epithelial carcinoma, borderline tumor, blastoma, and stromal tumor; esophageal cancers such as, but not limited to, squamous carcinoma, adenocarcinoma, adenoid cystic carcinoma, mucoepidermoid carcinoma, adenosquamous carcinoma, sarcoma, melanoma, plasmacytoma, verrucous carcinoma, and oat cell (small cell) carcinoma; gastric cancers such as, but not limited to, adenocarcinoma, mycosis (polypoid), ulceration, superficial spread, diffuse spread, malignant lymphoma, liposarcoma, fibrosarcoma, and carcinosarcoma; colon cancer; rectal cancer; liver cancer such as, but not limited to, hepatocellular carcinoma and hepatoblastoma; gallbladder cancer, such as adenocarcinoma; cholangiocarcinoma such as, but not limited to, papillary, nodular, and diffuse; lung cancer such as non-small cell lung cancer, squamous cell carcinoma (epidermoid carcinoma), adenocarcinoma, large cell carcinoma and small cell lung cancer; testicular cancers such as, but not limited to, germ cell tumor, seminoma, anaplastic, classical (typical), sperm cell cancer, non-seminoma, embryonal carcinoma, teratoma, choriocarcinoma (yolk sac tumor), prostate cancers such as, but not limited to, prostate intraepithelial neoplasia, adenocarcinoma, leiomyosarcoma, and rhabdomyosarcoma; pineal cancer; oral cancer such as, but not limited to, squamous cell carcinoma; basal carcinoma; salivary gland cancers such as, but not limited to, adenocarcinoma, mucoepidermoid carcinoma, and adenoid cystic carcinoma; pharyngeal cancers such as, but not limited to, squamous cell carcinoma and verrucous; skin cancers such as, but not limited to, basal cell carcinoma, squamous cell carcinoma and melanoma, superficial invasive melanoma, nodular melanoma, nevus-like malignant melanoma, acropigmented melanoma; kidney cancers such as, but not limited to, renal cell carcinoma, adenocarcinoma, suprarenal adenoid tumor, fibrosarcoma, transitional cell carcinoma (renal pelvis and/or uterus); wilms' tumor; bladder cancer such as, but not limited to, transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, carcinosarcoma. In addition, cancers include myxosarcoma, osteogenic sarcoma, endothelial sarcoma, lymphangial endothelial sarcoma, mesothelioma, synovioma, hemangioblastoma, epithelial carcinoma, cystadenocarcinoma, bronchogenic carcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, and papillary adenocarcinoma (for a review of such conditions, see Fishman et al, 1985, Medicine, 2 nd edition, J.B. Lippincott Co., Philadelphia, and Murphy et al, 1997, informational Decisions: The compact Book of Cancer Diagnosis, Treatment, and Recovery, Viking Penguin, Penguin Books U.S. A., Inc., United States of America).

The MSLN binding proteins of the present disclosure may also be used to treat or prevent a variety of cancers or other hyperproliferative diseases, including (but not limited to) the following: cancers, including bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid, and skin cancers; including squamous cell carcinoma; hematologic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, burkitt's lymphoma; hematological tumors of the myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; other tumors, including melanoma, seminoma, tetra carcinoma, neuroblastoma, and glioma; tumors of the central and peripheral nervous system, including astrocytomas, neuroblastoma, glioma, and schwannoma; tumors of mesenchymal origin, including fibrosarcoma, rhabdomyoma, and osteosarcoma; and other tumors, including melanoma, xeroderma pigmentosum, keratoma, seminoma, thyroid follicular cancer, and teratoma. It is also contemplated that cancers caused by aberrant apoptosis may also be treated by the methods and compositions of the present disclosure. Such cancers may include, but are not limited to, follicular lymphoma, cancers with the p53 mutation, hormone-dependent tumors of the breast, prostate and ovary, and precancerous lesions such as familial adenomatous polyposis and myelodysplastic syndrome. In particular embodiments, malignant or dysplastic changes (such as metaplasia and dysplasia) or hyperproliferative diseases are treated or prevented in the skin, lung, colon, breast, prostate, bladder, kidney, pancreas, ovary or uterus. In other specific embodiments, sarcoma, melanoma, or leukemia is treated or prevented.

In some embodiments, "treatment" or "treating" as used herein refers to a therapeutic treatment, wherein the aim is to slow down (alleviate) an undesired physiological condition, disorder or disease, or to obtain a beneficial or desired clinical result. For the purposes described herein, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; a reduction in the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the condition, disorder or disease state; delayed onset or slowed progression of the condition, disorder or disease; amelioration of a condition, disorder or disease state; and detectable or undetectable alleviation (partial or total) or enhancement or amelioration of the condition, disorder or disease. Treatment involves eliciting a clinically significant response without excessive levels of side effects. Treatment also includes extending survival compared to survival expected in the absence of treatment. In other embodiments, "treatment" or "treating" refers to a prophylactic measure, wherein the objective is to delay the onset of, or reduce the severity of, an undesirable physiological condition, disorder or disease, for example, in a person predisposed to the disease (e.g., an individual carrying a genetic marker for a disease such as breast cancer).

In some embodiments of the methods described herein, a MSLN binding protein as described herein is administered in combination with an agent for treating a particular disease, disorder, or condition. Agents include, but are not limited to, therapies involving antibodies, small molecules (e.g., chemotherapeutic drugs), hormones (steroids, peptides, etc.), radiation therapy (targeted delivery of gamma rays, X-rays, and/or radioisotopes, microwaves, UV radiation, etc.), gene therapy (e.g., antisense, retroviral therapy, etc.), and other immunotherapies. In some embodiments, the MSLN binding proteins described herein are administered in combination with an antidiarrheal agent, an antiemetic agent, an analgesic, an opioid, and/or a non-steroidal anti-inflammatory agent. In some embodiments, the MSLN binding proteins described herein are administered in combination with an anti-cancer agent. Non-limiting examples of anti-cancer agents that may be used in various embodiments of the present disclosure (including pharmaceutical compositions and dosage forms and kits of the present disclosure) include: acivicin; aclarubicin; (ii) aristozole hydrochloride; (ii) abelmoscine; (ii) Alexanox; aldesleukin; altretamine; an apramycin; amenthraquinone acetate; aminoglutethimide; amsacrine; anastrozole; an atramycin; an asparaginase enzyme; a triptyline; azacitidine; azatepa; (ii) azomycin; batimastat; benzotepa; bicalutamide; bisantrene hydrochloride; bisnefaede dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; briprimine; busulfan; actinomycin C; (ii) carroterone; a carbimide; a carbapenem; carboplatin; carmustine; a doxorubicin hydrochloride; folding to get new; cediogo, and cediogo; chlorambucil; a sirolimus; cisplatin; cladribine; krestist mesylate; cyclophosphamide; cytarabine; dacarbazine; actinomycin D; daunorubicin hydrochloride; decitabine; (ii) dexomaplatin; tizanoguanine; dizyguanine mesylate; diazaquinone;docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; drotandrosterone propionate; daptomycin; edatrexae; efluoronitine hydrochloride; elsamitrucin; enloplatin; an enpu urethane; epinastine; epirubicin hydrochloride; (ii) ebuzole; isosbacin hydrochloride; estramustine; estramustine sodium phosphate; etanidazole; etoposide; etoposide phosphate; etophenine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; (iii) flucitabine; a phosphorus quinolone; fostrexasin sodium; gemcitabine; gemcitabine hydrochloride; a hydroxyurea; idarubicin hydrochloride; ifosfamide; ilofovir dipivoxil; interleukin II (including recombinant interleukin II or rIL 2); interferon alpha-2 a; interferon alpha-2 b; interferon alpha-n 1; interferon alpha-n 3; interferon beta-I a; interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprorelin acetate; liazole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; (ii) maxolone; maytansine; mechlorethamine hydrochloride; megestrol acetate; megestrol acetate; melphalan; (ii) a melanoril; mercaptopurine; methotrexate; methotrexate sodium; chlorpheniramine; meltupipide; mitodomide; mitocarcin (mitocarcin); mitorubin (mitocromin); mitoxantrone; martin filamentation; mitomycin; mitospirane culturing; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; a noggin; ormaplatin; oshuzuren; paclitaxel; a pemetrexed; a pelithromycin; pentazocine; pellomycin sulfate; cultivating phosphoramide; pipobroman; piposulfan; piroxantrone hydrochloride; (ii) a plicamycin; pramipexole; porfimer sodium; a podomycin; deltemustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazole furan rhzomorph; (ii) lybodenosine; ludwimine; safrog; safrog hydrochloride; semustine; octreozine; sodium stannate; a sparamycin; helical germanium hydrochloride; spiromustine; spiroplatinum; streptonigrin; streptozotocin; a sulfochlorophenylurea; a talithromycin; sodium tegafur; tegafur; tiloxanthraquinone hydrochloride; temoporfin; (ii) teniposide; a tiroxiron; a testosterone ester; (ii) a thiopurine; thioguanine; thiotepa; (ii) a thiazole carboxamide nucleoside; substitute pulling bundleClearing; toremifene citrate; triton acetate; triciribine phosphate; trimetrexate; tritrosa glucuronide; triptorelin; tobramzole hydrochloride; uramustine; uretipi; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vincristine sulfate; vinorelbine tartrate; vinblastine sulfate; vinzolidine sulfate; (ii) vorozole; zeniplatin; 1, neat setastine; zorubicin hydrochloride. Other examples of anti-cancer drugs include, but are not limited to: 20-epi-1, 25 dihydroxy vitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; an acylfulvene; adenosylpentanol; (ii) Alexanox; aldesleukin; ALL-TK antagonist; altretamine; amifostine; (ii) amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; an angiogenesis inhibitor; an antagonist D; an antagonist G; anrlex; anti-dorsal morphogenetic protein-1 (anti-dorsallizing morphogenetic protein-1); an antiandrogen; prostate cancer; an antiestrogen; an antineoplastic ketone; an antisense oligonucleotide; glycine alfacin; an apoptosis gene modulator; an apoptosis modulator; (ii) an allopurinic acid; ara-CDP-DL-PTBA; arginine deaminase; sinapine (asularnine); atamestan; amoxicillin; azithromycin (axinatatin) 1; azithromycin 2; azithromycin 3; azasetron; azatoxin; diazo-leucine; baccatin III derivatives; balanol; batimastat; a BCR/ABL antagonist; benzodihydrophenol (benzodioxins); benzoyl staurosporine (benzostaurosporine); a beta-lactam derivative; beta-alethine; betacylomycin B; betulinic acid; a bFGF inhibitor; bicalutamide; a bisantrene group; bis-aziridinyl spermine; (ii) bisnefarde; hyperbolic group a (bistetralene a); bizelesin; brefflate; briprimine; (iii) butobactam; buthionine sulfoximine; calcipotriol; calphos protein C; a camptothecin derivative; canarypox IL-2; capecitabine; carboxamide-amino-triazole; a carboxyamidotriazole; CaRest M3; CARN 700; a cartilage derived inhibitor; folding to get new; casein kinase Inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquineA quinoxaline sulfonamide; (ii) cicaprost; a cis-porphyrin; cladribine; clomiphene analogs; clotrimazole; colismycin A; colismycin B; combretastatin a 4; a combretastatin analog; a concanagen; crambescidin 816; krestist; nostoc 8; a nostoc a derivative; curve A; cyclopentaquinone; cycloplatin (cycloplatam); (ii) sequomycin; cytarabine octadecyl phosphate; a cytolytic factor; hexestrol phosphate (cytostatin); daclizumab; decitabine; dehydromembrane ecteinascidin B; deslorelin; dexamethasone; (ii) dexifosfamide; dexrazoxane; (ii) verapamil; diazaquinone; a sphingosine B; didox; diethyl norspermine; dihydro-5-azacytidine; 9-dihydrotaxol; dioxamycin (dioxamycin); diphenylspiromustine; docetaxel; policosanol; dolasetron; doxifluridine; droloxifene; dronabinol; a multi-kamicin SA; ebselen selenium; etokomustine; edifulin; edrecolomab (edrecolomab); eflornithine; elemene; ethirimuron fluoride; epirubicin; epristeride; an estramustine analogue; an estrogen agonist; an estrogen antagonist; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; degree of fraunhise; flutemastine; a flashterone; fludarabine; fludaunorubicin hydrochloride (fluoroauroruronium hydrochloride); fowler; 2, fulvestrant; fostrexed; fotemustine; gadolinium deuteroporphyrin (gadolinium texaphyrin); gallium nitrate; galocitabine; ganirelix; (ii) a gelatinase inhibitor; gemcitabine; a glutathione inhibitor; hepsulfam; modulation of protein; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; iloperidone; ilofovir dipivoxil; ilomastat; an imidazocridinone; imiquimod; immunostimulatory peptides; insulin-like growth factor-I receptor inhibitors; an interferon agonist; an interferon; an interleukin; iodobenzylguanidine; iododoxorubicin; 4-sweet potato picrol; iprop; isradine; isobongrezole; isohigh halichondrin B; itasetron; garcinolone acetonide (jasplakinolide); kahalard F (kahalalide F); lamellarin triacetate-N; lanreotide; a renamycin; leguminous kiosks; sulfuric acid lentinus edodes powderA sugar; a rebustatin; letrozole; leukemia inhibitory factor; leukocyte interferon-alpha; leuprorelin + estrogen + progesterone; leuprorelin; levamisole; liazole; a linear polyamine analog; a lipophilic glycopeptide; a lipophilic platinum compound; lissoclinamide 7; lobaplatin; earthworm phosphatide; lometrexol; lonidamine; losoxanthraquinone; HMG-CoA reductase inhibitors (such as but not limited to lovastatin, pravastatin, fluvastatin, statins, simvastatin, and atorvastatin); loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; a lytic peptide; maytansine; manostatin a; marimastat; (ii) maxolone; mammary silk arrestin; a matrix dissolution factor inhibitor; a matrix metalloproteinase inhibitor; (ii) a melanoril; mebarone (merbarone); avorelin; methioninase (methioninase); metoclopramide; an inhibitor of MIF; mifepristone; miltefosine; a Millisetil; mismatched double-stranded RNA; mitoguazone; dibromodulcitol; mitomycin analogs; mitonaphthylamine; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofagotine; moraxest; human chorionic gonadotropin monoclonal antibody; monophosphoryl lipid a + mycobacterial cell wall sk; mopidanol; multiple drug resistance gene inhibitors; multiple tumor suppressor 1-based therapy; mustard anticancer agent; indian sponge B (mycaperoxide B); a mycobacterial cell wall extract; myriaporone; n-acetyldinaline; an N-substituted benzamide; nafarelin; nagestip; naloxone + pentazocine; napavin; naphterpin; a nartostim; nedaplatin; nemorubicin; neridronic acid; a neutral endopeptidase; nilutamide; nisamycin; a nitric oxide modulator; a nitroxide antioxidant; nitrulyn; o6-benzylguanine; octreotide; okicenone; an oligonucleotide; onapristone; ondansetron; ondansetron; oracin; an oral cytokine inducer; ormaplatin; an oxateclone; oxaliplatin; oxanonomycin; paclitaxel; a paclitaxel analog; a paclitaxel derivative; palauamine; palmitoylrhizoxin; pamidronic acid; panaxatriol; panomifen; a parabencin; pomagliptin; a pemetrexed; pedasine (peldesine); sodium pentosan polysulfate; pentostatin; spraying trozole; perfluor bromoalkane(ii) a Cultivating phosphoramide; perilla alcohol; phenazinomycin (phenazinomomycin); a salt of phenylacetic acid; a phosphatase inhibitor; streptolysin preparations (picibanil); pilocarpine hydrochloride; pirarubicin; pirtroxine; placetin A; placetin B; a plasminogen activator inhibitor; a platinum complex; a platinum compound; a platinum-triamine complex; porfimer sodium; a podomycin; prednisone; propyl bisacridone; prostaglandin J2; a proteasome inhibitor; protein a-based immunomodulators; inhibitors of protein kinase C; inhibitors of protein kinase C; microalgae; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurin; methoxypyrazoline acridine; pyridoxylated hemoglobin polyoxyethylene conjugates; a raf antagonist; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; (ii) a ras inhibitor; ras-GAP inhibitors; demethylated reteplatin; rhenium Re 186 etidronate; rhizomycin; a ribozyme; RII isotretinoin amide; ludwimine; roxitukale; romurtide; loquimex; rubiginone B1; ruboxyl; safrog; saintopin; SarCNU; inositol A; sargrastim; a Sdi 1 mimetic; semustine; senescence-derived inhibitor 1; a sense oligonucleotide; a signal transduction inhibitor; a signal transduction modulator; a single-chain antigen-binding protein; a texaphyrin; sobuconazole; sodium boron carbonate; sodium phenylacetate; solverol; a growth regulator binding protein; sonaming; phosphono-winteric acid; spicamycin d (spicamycin d); spiromustine; (ii) spandex; spongistatin 1(spongistatin 1); squalamine; a stem cell inhibitor; inhibitors of stem cell division; stiiamide; a matrix-dissolving protease inhibitor; sulfinosine; a potent vasoactive intestinal peptide antagonist; (ii) surfasta; suramin; swainsonine; a synthetic glycosaminoglycan; tamustine; tamoxifen methiodide; taulomustine; tazarotene; sodium tegafur; tegafur; telluropyrylium; a telomerase inhibitor; temoporfin; temozolomide; (ii) teniposide; tetrachlorodecaoxide (tetrachlorodecaoxide); tetrazomine; somatic embryo element (thalblastitin); thiocoraline (thiocoraline); thrombopoietin; a thrombopoietin mimetic; thymalfasin (Thymalfasin); a thymopoietin receptor agonist; thymotreonam; thyroid stimulating hormone; the purpurin BTin ethyl ethylpururin; tirapazamine; titanocene dichloride (titanocene bichloride); topstein; toremifene; a totipotent stem cell factor; a translation inhibitor; tretinoin; triacetyl uridine; triciribine; trimetrexate; triptorelin; tropisetron; tolite; tyrosine kinase inhibitors; tyrosine phosphorylation inhibitors (tyrphostin); an UBC inhibitor; ubenimex; urogenital sinus-derived growth inhibitory factor; a urokinase receptor antagonist; vapreotide; variolin B; a carrier system; erythrocyte gene therapy; vilareol; veratramine; verdins; verteporfin; vinorelbine; vinxaline;vorozole; zanoteron; zeniplatin; benzalvitamin c (zilascorb); and neat stastatin ester. Other anticancer drugs are 5-fluorouracil and folinic acid. Both agents are particularly useful when used in methods employing thalidomide and a topoisomerase inhibitor. In some embodiments, the anti-MSLN single domain binding proteins of the present disclosure are used in combination with gemcitabine.

In some embodiments, the MSLN binding proteins described herein are administered before, during, or after surgery.

Method for detecting mesothelin expression and diagnosis of mesothelin-associated cancer

According to another embodiment of the present disclosure, a kit for detecting mesothelin expression in vitro or in vivo is provided. The kit comprises the aforementioned MSLN binding protein (e.g., a labeled anti-MSLN single domain antibody or antigen-binding fragment thereof), and one or more compounds for detecting the label. In some embodiments, the label is selected from the group consisting of a fluorescent label, an enzymatic label, a radioactive label, a nuclear magnetic resonance active label, a luminescent label, and a chromophore label.

In some cases, mesothelin expression is detected in a biological sample. The sample can be any sample, including but not limited to tissue from biopsies, autopsies, and pathological specimens. Biological samples also include tissue sections, e.g., frozen sections for histological purposes. Biological samples also include bodily fluids such as blood, serum, plasma, sputum, spinal fluid, or urine. The biological sample is typically obtained from a mammal, such as a human or non-human primate.

In one embodiment, a method is provided for determining whether a subject has cancer by contacting a sample from the subject with an anti-MSLN single domain antibody disclosed herein and detecting binding of the single domain antibody to the sample. An increase in binding of the antibody to the sample as compared to binding of the antibody to the control sample identifies the subject as having cancer.

In another embodiment, a method is provided for confirming a diagnosis of cancer in a subject by contacting a sample from a subject diagnosed with cancer with an anti-MSLN single domain antibody disclosed herein and detecting binding of the antibody to the sample. An increase in binding of the antibody to the sample compared to binding of the antibody to the control sample confirms diagnosis of the cancer in the subject.

In some examples of the disclosed methods, the single domain antibody is directly labeled.

In some examples, the method further comprises contacting a second antibody that specifically binds to the single domain antibody with the sample; and detecting binding of the second antibody. An increase in binding of the second antibody to the sample, as compared to binding of the second antibody to a control sample, detects or confirms a diagnosis of cancer in the subject.

In some cases, the cancer is mesothelioma, prostate cancer, lung cancer, gastric cancer, squamous cell carcinoma, pancreatic cancer, cholangiocarcinoma, triple negative breast cancer, or ovarian cancer, or any other type of cancer that expresses mesothelin.

In some examples, the control sample is a sample from a subject without cancer. In a particular example, the sample is a blood or tissue sample.

In some cases, an antibody that binds (e.g., specifically binds) to mesothelin is directly labeled with a detectable label. In another embodiment, the antibody that binds (e.g., specifically binds) to mesothelin (the first antibody) is unlabeled, while the second antibody or other molecule that can bind to an antibody that specifically binds mesothelin is labeled. The second antibody is selected to be capable of specifically binding to the specific species and class of the first antibody. For example, if the first antibody is llama IgG, the second antibody may be anti-llama IgG. Other molecules that can bind to antibodies include, but are not limited to, protein a and protein G, both of which are commercially available. Suitable labels for the antibody or second antibody are as described above and include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, magnetic agents and radioactive materials. Non-limiting examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase. Non-limiting examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin. Non-limiting examples of suitable fluorescent materials include umbelliferone, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin. A non-limiting exemplary luminescent material is luminol; a non-limiting exemplary magnetic agent is gadolinium, and non-limiting exemplary radioactive labels include 125I, 131I, 35S, or 3H.

In an alternative embodiment, mesothelin may be determined in a biological sample by a competitive immunoassay that employs a mesothelin standard labeled with a detectable substance and an unlabeled antibody that specifically binds mesothelin. In this assay, a biological sample, a labeled mesothelin standard and an antibody that specifically binds mesothelin are mixed together and the amount of labeled mesothelin standard bound to the unlabeled antibody is determined. The amount of mesothelin in the biological sample is inversely proportional to the amount of labeled mesothelin standard bound to the antibody that specifically binds mesothelin.

The immunoassays and methods disclosed herein can be used for a variety of purposes. In one embodiment, an antibody that specifically binds to mesothelin may be used to detect mesothelin production in cells in cell culture. In another embodiment, the antibody can be used to detect the amount of mesothelin in a biological sample, such as a tissue sample or a blood or serum sample. In some examples, the mesothelin is cell surface mesothelin. In other examples, the mesothelin is soluble mesothelin (e.g., mesothelin in a cell culture supernatant or mesothelin in a bodily fluid sample such as a blood or serum sample.

In one embodiment, a kit for detecting mesothelin in a biological sample, such as a blood sample or a tissue sample, is provided. For example, to confirm a diagnosis of cancer in a subject, a biopsy may be performed to obtain a tissue sample for histological examination. Alternatively, a blood sample may be taken to detect the presence of soluble mesothelin protein or fragments. In accordance with the present disclosure, a kit for detecting a polypeptide will typically comprise a single domain antibody that specifically binds mesothelin. In some embodiments, an antibody fragment, such as a scFv fragment, a VH domain, or a Fab, is included in the kit. In further embodiments, the antibody is labeled (e.g., with a fluorescent, radioactive, or enzymatic label).

In one embodiment, the kit includes instructional materials that disclose the means for using antibodies that bind mesothelin. The instructional material may be written in electronic form (e.g., a computer diskette or compact disk) or visual form (e.g., a video file). The kit may also include other components to facilitate the particular application for which the kit is designed. Thus, for example, the kit may additionally comprise means for detecting the label (e.g.an enzyme substrate for an enzymatic label, a filter set for detecting a fluorescent label, a suitable secondary label, such as a secondary antibody, etc.). The kit may additionally include buffers and other reagents conventionally used to carry out particular methods. Such kits and suitable contents are well known to those skilled in the art.

In one embodiment, the diagnostic kit comprises an immunoassay. Although the details of the immunoassay may vary depending on the particular format employed, the method of detecting mesothelin in a biological sample generally comprises the step of contacting the biological sample with an antibody that specifically reacts with a mesothelin polypeptide under immunoreactive conditions. The antibodies are allowed to bind specifically under immunoreactive conditions to form an immune complex, and the presence of the immune complex (bound antibody) is detected, either directly or indirectly.

Methods for determining the presence or absence of cell surface markers are well known in the art. For example, the antibody may be conjugated to other compounds including, but not limited to, enzymes, magnetic beads, colloidal magnetic beads, haptens, fluorochromes, metal compounds, radioactive compounds, or drugs. The antibodies may also be used in immunoassays, such as, but not limited to, Radioimmunoassays (RIA), ELISA, or immunohistochemistry assays. Antibodies can also be used for Fluorescence Activated Cell Sorting (FACS). FACS uses multiple color channels, low and obtuse angle light scattering detection channels, and impedance channels, as well as other more complex detection levels, to separate or sort cells (see U.S. patent No. 5,061,620). As disclosed herein, any single domain antibody that binds mesothelin can be used in these assays. Thus, these antibodies can be used in conventional immunoassays, including but not limited to ELISA, RIA, FACS, tissue immunohistochemistry, Western blotting, or immunoprecipitation.

The present invention provides embodiments including, but not limited to:

1. a single domain mesothelin binding protein wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 41, 42, 43 or 44 or comprising one or more amino acid residue substitutions with respect to SEQ ID NO 41, 42, 43 or 44.

2. The single domain mesothelin binding protein according to embodiment 1, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:41 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 41.

3. The single domain mesothelin binding protein according to embodiment 1 or 2, wherein said protein comprises a sequence comprising a conserved region comprising a sequence that is identical to SEQ ID NO:42 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 42.

4. The single domain mesothelin binding protein according to embodiment 1,2 or 3, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:43 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 43.

5. The single domain mesothelin binding protein according to embodiment 1,2 or 3, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:44 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 44.

6. The single domain mesothelin binding protein according to embodiment 1, wherein said protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO: 41; (ii) a stretch of amino acids corresponding to SEQ ID NO 42; (iii) a stretch of amino acids corresponding to SEQ ID NO 43; and (iv) a stretch of amino acids corresponding to SEQ ID NO: 44.

7. A single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

8. The single domain mesothelin binding protein according to any one of embodiments 1-7, wherein said protein comprises a sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID NOs 1-40, 58 and 60-62.

9. The single domain mesothelin binding protein according to any one of embodiments 1-8, wherein said protein comprises one or more modifications that result in humanization of said binding protein.

10. The single domain mesothelin binding protein of embodiment 9, wherein said modification comprises a substitution, addition or deletion of an amino acid residue.

11. The single domain mesothelin binding protein according to any one of embodiments 1-10, wherein said protein comprises between 111 amino acids and 124 amino acids.

12. The single domain mesothelin binding protein according to any one of embodiments 1-11, wherein said protein comprises a VHH domain derived from a non-human source.

13. The single domain mesothelin-binding protein according to any one of embodiments 1-12, wherein said protein comprises a llama VHH domain.

14. The single domain mesothelin binding protein according to any one of embodiments 1 to 13, wherein said protein binds to an epitope of mesothelin, wherein said epitope is located in region I comprising amino acid residues 296-390 of SEQ ID NO:57, region II comprising amino acid residues 391-486 of SEQ ID NO:57 or region III comprising amino acid residues 487-598 of SEQ ID NO: 57.

15. A single domain mesothelin binding protein, wherein said protein comprises one or more conserved regions comprising a sequence identical to SEQ ID NO 45, 46, 47, 48, 49 or 50 or comprising one or more amino acid residue substitutions with respect to SEQ ID NO 45, 46, 47, 48, 49 or 50.

16. The single domain mesothelin binding protein of embodiment 15, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:45 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 45.

17. The single domain mesothelin binding protein of embodiment 15 or 16, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:46 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 46.

18. The single domain mesothelin binding protein of embodiment 15, 16 or 17, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:47 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 47.

19. The single domain mesothelin binding protein according to any one of claims 15-18, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:48 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 48.

20. The single domain mesothelin binding protein according to any one of claims 15-19, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:49 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 49.

21. The single domain mesothelin binding protein according to any one of embodiments 15-20, wherein said protein comprises a conserved region comprising a sequence that is identical to SEQ ID NO:50 or comprises one or more amino acid residue substitutions relative to SEQ ID NO: 50.

22. The single domain mesothelin binding protein according to embodiment 1, wherein said protein comprises (i) a stretch of amino acids corresponding to SEQ ID NO: 45; (ii) a stretch of amino acids corresponding to SEQ ID NO 46; (iii) a stretch of amino acids corresponding to SEQ ID NO. 47; (iv) a stretch of amino acids corresponding to SEQ ID NO 48; (v) a stretch of amino acids corresponding to SEQ ID NO. 49; and (vi) a stretch of amino acids corresponding to SEQ ID NO: 50.

23. A single domain mesothelin binding protein, wherein said protein comprises the formula:

f1-r1-f2-r2-f3-r3-f4

24. The single domain mesothelin binding protein according to any one of embodiments 15-23, wherein said protein comprises a sequence that is at least 80% identical to a sequence selected from the group consisting of SEQ ID Nos 30-40, 58 and 60-62.

25. The single domain mesothelin binding protein according to any one of embodiments 15-24, wherein said protein comprises from 111 amino acids to 119 amino acids.

26. The single domain mesothelin binding protein according to any one of claims 15-25, wherein said protein comprises a VHH domain derived from a non-human source.

27. The single domain mesothelin-binding protein according to any one of claims 15-26, wherein said protein comprises a llama VHH domain.

28. The single domain mesothelin binding protein according to any one of embodiments 15-27, wherein said protein binds to a human mesothelin protein comprising the sequence shown in SEQ ID NO: 57.

29. The single domain mesothelin binding protein according to any one of embodiments 15 to 28, wherein said protein binds to an epitope of mesothelin, wherein said epitope is in region I comprising amino acid residues 296-390 of SEQ ID NO:57, region II comprising amino acid residues 391-486 of SEQ ID NO:57 or region III comprising amino acid residues 487-598 of SEQ ID NO: 57.

30. The single domain mesothelin binding protein according to any one of embodiments 1-29, wherein said binding protein is a chimeric antibody or a humanized antibody.

31. The single domain binding protein according to any one of embodiments 1-30, wherein said binding protein is a single domain antibody.

32. The single domain binding protein according to any one of claims 15-31, wherein said binding protein is a humanized single domain antibody.

33. The single domain binding protein according to any one of embodiments 1-32, wherein said binding protein comprises the sequence set forth in SEQ ID NO: 58.

34. A polynucleotide encoding a single domain mesothelin binding protein according to any one of embodiments 1-33.

35. A vector comprising the polynucleotide according to embodiment 34.

36. A host cell transformed with the vector according to embodiment 35.

37. A pharmaceutical composition comprising (i) a single domain mesothelin-binding protein according to any one of embodiments 1-33, a polynucleotide according to embodiment 33, a vector according to embodiment 35 or a host cell according to embodiment 36, and (ii) a pharmaceutically acceptable carrier.

38. A method of producing a single domain mesothelin binding protein according to any one of embodiments 1-33, said method comprising culturing a host transformed or transfected with a vector comprising a nucleic acid sequence encoding a single domain mesothelin binding protein according to any one of embodiments 1-33 under conditions that allow expression of the mesothelin binding protein and recovery and purification of the produced protein from the culture.

39. A method of treating or ameliorating a hyperplastic or neoplastic disease comprising administering to a subject in need thereof a mesothelin binding protein according to any one of embodiments 1-33.

40. The method of embodiment 39, wherein the subject is a human.

41. The method of embodiment 40, wherein the method further comprises administering an agent in combination with the single domain mesothelin binding protein according to any one of embodiments 1-33.

42. The method of any one of embodiments 39-41, wherein said single domain mesothelin binding protein selectively binds to mesothelin-expressing tumor cells.

43. The method of embodiment 42, wherein said single domain mesothelin binding protein mediates killing of mesothelin-expressing tumor cells by T cells.

44. The method of any one of embodiments 39-43 wherein the neoplastic disease comprises a solid tumor disease.

45. The method of embodiment 44, wherein the solid tumor disease comprises mesothelioma, lung cancer, gastric cancer, ovarian cancer, or triple negative breast cancer.

46. The method of embodiment 45, wherein the solid tumor disease is metastatic.

47. A single domain mesothelin binding protein, wherein said protein comprises one or more CDRs selected from the group consisting of SEQ ID Nos. 51-56 and 63-179.

48. The single domain mesothelin binding protein of embodiment 47, wherein said protein comprises CDR1, which CDR1 comprises the sequence set forth in any one of SEQ ID Nos. 51, 54 and 63-101.

49. The single domain mesothelin binding protein according to embodiment 47 or 48, wherein said protein comprises CDR2, which CDR2 comprises the sequence set forth in any one of SEQ ID Nos. 52, 55 and 102-140.

50. The single domain mesothelin binding protein according to any one of embodiments 47-49, wherein said protein comprises CDR3, which CDR3 comprises the sequence set forth in any one of SEQ ID Nos. 53, 56 and 141 and 179.

51. The single domain mesothelin binding protein according to any of embodiments 47-50, wherein said protein comprises framework region 1(f1), which framework region 1 comprises the sequence as set forth in any of SEQ ID No. 180 and 218.

52. The single domain mesothelin binding protein according to any of embodiments 47-51, wherein said protein comprises framework region 2(f2), which framework region 2 comprises the sequence as set forth in any of SEQ ID No. 219 and 257.

53. The single domain mesothelin binding protein according to any of embodiments 47-52, wherein said protein comprises framework region 3(f3), which framework region 3 comprises the sequence as set forth in any of SEQ ID No. 258-.

54. The single domain mesothelin binding protein according to any of embodiments 47-53, wherein said protein comprises framework region 4(f4), which framework region 4 comprises the sequence as set forth in any of SEQ ID No. 297-335.

55. The single domain mesothelin binding protein according to any one of embodiments 47-54, wherein said protein comprises the amino acid sequence set forth in any one of SEQ ID Nos. 1-40 and 58.

56. A polynucleotide encoding the single domain mesothelin binding protein according to any one of embodiments 47-55.

57. A vector comprising the polynucleotide according to embodiment 56.

58. A host cell transformed with the vector according to embodiment 57.

59. A pharmaceutical composition comprising (i) a single domain mesothelin-binding protein according to any one of embodiments 47-55, a polynucleotide according to embodiment 56, a vector according to embodiment 57 or a host cell according to embodiment 58, and (ii) a pharmaceutically acceptable carrier.

60. A method of producing a single domain mesothelin binding protein according to any one of embodiments 47-55, said method comprising culturing a host transformed or transfected with a vector comprising a nucleic acid sequence encoding a single domain mesothelin binding protein according to any one of embodiments 47-55 under conditions that allow expression of the mesothelin binding protein and recovery and purification of the produced protein from the culture.

61. A method of treating or ameliorating a hyperplastic or neoplastic disease comprising administering to a subject in need thereof a mesothelin binding protein according to any one of embodiments 47-55.

62. The method of embodiment 61, wherein the subject is a human.

63. The method of embodiment 62, wherein the method further comprises administering an agent in combination with the single domain mesothelin binding protein of any one of embodiments 1-33.

64. The method of any one of embodiments 61-63, wherein said single domain mesothelin-binding protein selectively binds to mesothelin-expressing tumor cells.

65. The method of embodiment 64, wherein said single domain mesothelin binding protein mediates killing of mesothelin-expressing tumor cells by T cells.

66. The method according to any one of embodiments 61-65, wherein the neoplastic disease comprises a solid tumor disease.

67. The method of embodiment 66, wherein the solid tumor disease comprises mesothelioma, lung cancer, gastric cancer, ovarian cancer, or triple negative breast cancer.

68. The method of embodiment 67, wherein the solid tumor disease is metastatic.

Examples

The following examples further illustrate the described embodiments without limiting the scope of the invention.

Example 1: exemplary anti-MSLN single domain antibodies of the disclosure mediate T cell killing of mesothelin-expressing cancer cells Capacity of cells

Exemplary anti-MSLN single domain antibody sequences were transfected into Expi293 cells (Invitrogen). The amount of exemplary anti-MSLN antibody in conditioned media from transfected Expi293 cells was quantified using an Octet instrument with a protein a tip and a standard curve was made using control anti-MSLN antibody.

Titrations of conditioned media were added to a TDCC assay (T cell dependent cytotoxicity assay) to assess whether anti-MSLN single domain antibodies were able to synapse between T cells and the mesothelin-expressing ovarian cancer cell line OVCAR 8. After 48 hours the viability of OVCAR8 cells was measured. As can be seen, exemplary anti-MSLN single domain antibodies mediate T cell killing.

Furthermore, it can be seen that the TDCC activity of the exemplary anti-MSLN single domain antibody is specific for mesothelin-expressing cells, as this exemplary antibody does not mediate killing of T cells on LNCaP cells that do not express mesothelin.

Example 2: evaluation of several MSLN-targeted trispecific antigen binding containing MSLN binding domains according to the present disclosure Methods for binding and cytotoxic activity of proteins

Protein production

The sequence of the MSLN targeting trispecific molecule containing the MSLN binding protein according to the present disclosure was cloned into the mammalian expression vector pCDNA 3.4(Invitrogen) preceded by a leader sequence followed by a 6x histidine tag. Expi293F cells (Life Technologies A14527) were maintained in suspension at 0.2-8 × 1e6 cells/mL in Expi293 medium in Optimum Growth Flasks (Thomson). Purified plasmid DNA was transfected into Expi293 cells according to the Expi293 Expression System Kit (Life Technologies, A14635) protocol and maintained for 4-6 days after transfection. The amount of the exemplary trispecific protein tested from transfected Expi293 cells in conditioned media was quantified using an Octet instrument with a protein a tip and a standard curve was prepared using the control trispecific protein.

Cytotoxicity assays

The ability of T cell conjugates, including trispecific molecules, to direct T cell killing of tumor cells was measured using a human T cell dependent cytotoxicity (TDCC) assay (Nazarian et al 2015.J Biomol screen.20: 519-27). In this assay, T cells and target cancer cell line cells were mixed together in 384 well plates at a ratio of 10:1 and different amounts of the trispecific protein tested were added. Tumor cell lines were engineered to express luciferase protein. After 48 hours, Steady-Luminescent Assay(Promega)。

In this study, a titration of conditioned medium was added to the TDCC assay (T cell dependent cytotoxicity assay) to assess whether the anti-MSLN single domain antibody was able to synapse between T cells and the mesothelin expressing ovarian cancer cell line OVCAR 8. After 48 hours the viability of OVCAR8 cells was measured. It can be seen that the trispecific protein mediates T cell killing. FIG. 1 shows exemplary cell viability assays with the trispecific proteins tested 2A2 and 2A 4. The EC for TDCC activity of several other trispecific proteins tested are listed in Table 1 below₅₀。

Table 1: TDCC Activity of MSLN-Targeted trispecific proteins comprising MSLN-binding proteins according to the disclosure

Furthermore, it was observed that the TDCC activity of the tested trispecific protein targeting MSLN was specific for mesothelin expressing cells, as the tested trispecific protein did not mediate killing of T cells on LNCaP cells not expressing mesothelin. In particular, the trispecific proteins 2a2, 11F3, 9H2, 5C2, 10B3, 2F4, 5F2, 7F1, 2F4, 5H1, 3B4 and 7H2 did not show any TDCC activity on LnCaP cells.

Example 3: ADCC Activity of exemplary anti-MSLN Single Domain antibodies of the present disclosure

This study was intended to determine the ability of exemplary anti-MSLN single domain antibodies of the present disclosure to mediate ADCC as compared to comparative llama anti-MSLN antibodies without sequence modifications or substitutions as exemplary antibodies of the present disclosure. Both antibodies are expressed as multidomain proteins comprising additional immunoglobulin domains.

Material

The donors were subjected to leukapheresis and NK cells were isolated from the leukocyte packet (leukapack) by a cell purification group using the Milteni AutoMacs Pro negative selection system. NK cells were maintained at 4 ℃ overnight on a shaker, then washed, counted and counted at 4X 10⁶Individual cells/mL were resuspended in complete RPMI for ADCC assay.

Target: tumor cell targets were selected based on mesothelin expression. Targets were washed and counted. Will be 6X 10⁶Each target was resuspended IN complete RPMI and at a final concentration of 10. mu.M calcein (Sigma # C1359-00UL CALCEIN AM 4MM IN ANHYDROUS DMSO) at 37 ℃, 5% CO₂Mark down for 40 minutes. Cells were washed twice in PBS, resuspended in complete RPMI, and incubated at 37 deg.C with 5% CO₂Incubate for 2 hours. After labeling, the target cells were washed, recounted, and counted at 0.2X 10⁶The concentration of individual cells/mL was resuspended in complete RPMI for ADCC assay.

Method

ADCC assays were performed in 96-well round bottom tissue culture plates (Corning 3799). The test protein was titrated from 20. mu.g/mL to 0.0002. mu.g/mL by including 10. mu.L (1:10 dilution) in 1000. mu.L of complete RPMI containing 10% FCS. 50 μ L of calcein-labeled target was added to contain 10,000 cells. Target cells and various concentrations of multidomain proteins containing exemplary anti-MSLN single domain antibodies or comparative antibodies were incubated at 4 ℃ for 40 minutes, followed by the addition of 50 μ L of NK cell effector to contain 100,000 cells (10:1E: T ratio). The cultures were incubated at 37 ℃ for 4 hours, then the supernatant was removed and the release of calcein was determined by measuring the fluorescence at 485-. The 100% lysis value was determined by lysis of six wells of labeled target with Igepal 630 detergent (3 μ Ι _ per well) and the spontaneous lysis value was determined by measuring the fluorescence in the supernatant from the individual targets.

Statistical analysis

The percent (%) specific lysis was defined as (sample fluorescence) - (auto-lysis fluorescence)/(100% lysis-auto-lysis fluorescence). Spontaneous lysis was determined by wells containing only the target, whereas 100% lysis was determined by wells lysing the target with IGEPAL CA 630 detergent. Raw data were entered into an Excel spreadsheet with a formula embedded to calculate% specific lysis and the resulting values were transferred to a graphical program (GraphPad Prism) where the data were converted to a curve fit. Subsequent analysis (linear regression calculation) was then performed in GraphPad to generate ECs₅₀The value is obtained.

Results and discussion

Effector NK cells in wells incubated with multidomain proteins containing comparative anti-MSLN antibodies were unable to mediate killing of calcein labeled target cells, while effectors in wells with multidomain proteins containing exemplary anti-MSLN single domain antibodies of the present disclosure were able to mediate antibody-dependent cellular cytotoxicity as measured by specific lytic activity (% specific lysis).

Conclusion

Exemplary anti-MSLN single domain antibodies of the present disclosure mediate significantly higher levels of killing of mesothelin-expressing target cells compared to comparative llama anti-MSLN single domain antibodies without sequence substitutions, modifications, or humanizations.

Example 4: CDC activity of exemplary anti-MSLN single domain antibodies of the disclosure

To evaluate the anti-tumor activity of exemplary anti-MSLN single domain antibodies according to the present disclosure on cancer cells, cytotoxic activity in the a431/H9 and NCI-H226 cell models was tested in the presence of human serum as a source of complement. Exemplary anti-MSLN single domain antibodies are expressed as multidomain proteins containing additional immunoglobulin domains. It can be seen that multidomain proteins containing exemplary anti-MSLN single domain antibodies of the present disclosure exert potent CDC activity by killing about 40% of the a431/H9 and more than 30% of the NCI-H226 mesothelioma cell line, and show no activity on the mesothelin-negative a431 cell line. Comparative llama anti-MSLN antibodies without sequence modifications or substitutions as the exemplary antibodies of the present disclosure did not show activity at the same concentrations.

To analyze the role of complement in the anti-tumor activity of exemplary anti-MSLN single domain antibodies, flow cytometry was used to determine the binding of C1q to Cancer cells reactive with anti-mesothelin human mabs, following the maturation protocol used to characterize rituximab, ofatumumab and other anti-CD 20 therapeutic mabs (Pawluczkowycz et al, J Immunol 183: 749-bricks 758, 2009; Li et al, Cancer Res 68: 2400-bricks 2408, 2008). It has been previously shown that, like MORAB-009, HN1 human mAb, which is specific for the mesothelin domain I (away from the cell surface), does not exhibit any CDC activity on mesothelin-expressing Cancer cells (Ho et al, Int J Cancer 128: 2020) -2030, 2011).

However, it can be seen that C1q complement binds to A431/H9 or NCI-H226 cells in the presence of exemplary anti-MSLN single domain antibodies. In contrast, no C1q binding was found in the presence of the comparative llama anti-MSLN antibody. Furthermore, binding of C1q to cancer cells correlated in a dose-responsive manner with cellular binding of exemplary anti-MSLN single domain antibodies. These results demonstrate that exemplary anti-MSLN single domain antibodies exhibit improved CDC activity relative to comparative llama anti-MSLN antibodies.

Example 5: MSLN-targeted trispecific antigen binding containing an MSLN binding domain (MH6T) according to the present disclosure Protein-directed T cell killing of MSLN-expressing ovarian cancer cells

The ability of T cell conjugates, including trispecific molecules, to direct T cell killing of tumor cells was measured using a human T cell dependent cytotoxicity (TDCC) assay (Nazarian et al 2015.J Biomol screen.20: 519-27). The Caov3 cells used in this assay were engineered to express luciferase. T cells from 5 different healthy donors (donor 02, donor 86, donor 41, donor 81 and donor 34) and the target cancer cell Caov3 were mixed together, different amounts of MSLN-targeting trispecific antigen-binding protein (SEQ ID NO:58) containing the MSLN binding domain (MH6T) were added, and the mixture was incubated at 37 ℃ for 48 hours. Caov3 cells and T cells were also incubated with the control trispecific molecule GFP TriTAC (SEQ ID NO:59) targeting GFP for 48 hours at 37 ℃. After 48 hours, the remaining viable tumor cells were quantified by luminescence assay.

It was observed that MSLN-targeted trispecific antigen-binding proteins containing the MSLN binding domain (MH6T) were able to direct T cells from all 5 healthy donors to kill the target cancer cell, Caov3 (as shown in figure 2), whereas the control GFP TriTAC molecule was unable to direct T cells from any of these 5 healthy donors to kill the Caov3 cells (also shown in figure 2).

Further assays were performed using OVCAR3 cells using the same protocol as above. It was observed that MSLN-targeted trispecific antigen-binding proteins containing the MSLN binding domain (MH6T) were able to direct T cells from all 5 healthy donors to kill the target cancer cell OVCAR3 (as shown in figure 3), whereas the control GFP TriTAC molecule was unable to direct T cells from any of these 5 healthy donors to kill OVCAR3 cells (also shown in figure 3).

EC for killing target cells expressing MSLN are listed in Table II below₅₀The value is obtained.

Table II: MSLN-targeting trispecific antigen-binding proteins (containing the MH6T domain) directed EC killing of MSLN-expressing ovarian cancer cell lines by T cells from 5 different healthy donors₅₀The value is obtained. Representative illustrations of the raw data are provided in fig. 2 and 3.

Example 6: MSLN-targeted trispecific antigen binding containing an MSLN binding domain (MH6T) according to the present disclosure The protein directs T cells to kill cells expressing MSLN, but does not direct killing of cells not expressing MSLN

In this assay, T cells from healthy donors were incubated with target cancer cells expressing MSLN (Caov3 cells, Caov4 cells, OVCAR3 cells, and OVCAR8 cells) or target cancer cells not expressing MSLN (NCI-H510A cells, MDAPCa2b cells). Each target cell used in this study was engineered to express luciferase. Different amounts of MSLN targeting trispecific antigen-binding protein containing the MH6T domain (SEQ ID NO:58) were added to the above listed mixture of T cells and target cancer cells. The mixture was incubated at 37 ℃ for 48 hours. After 48 hours, the remaining viable target cancer cells were quantified using a luminescence assay.

It was observed that MH6T domain containing MSLN-targeted trispecific antigen-binding proteins were able to direct T cells to kill target cancer cells expressing MSLN (i.e., Caov3, Caov4, OVCAR3, and OVCAR8 cells, as shown in fig. 4). However, the MSLN-targeting trispecific antigen-binding protein containing the MH6T domain was unable to direct T cells to kill target cancer cells that do not express MSLN (MDAPCa2b and NCI-H510A cells), as also shown in FIG. 4.

EC for killing MSLN expressing cancer cells are listed in Table III below₅₀The value is obtained.

Table III: trispecific antigen-binding proteins targeting MSLN (containing MH6T domain) direct T cell killing of EC of MSLN-expressing cancer cell lines₅₀The value is obtained.

Example 7: MSLN-targeted trispecific antigen binding containing an MSLN binding domain (MH6T) according to the present disclosure Protein-directed T cell killing of human ovarian carcinoma cell lines from cynomolgus monkeys

In this assay, peripheral blood mononuclear cells (PBMC; T cells are part of PBMC) from cynomolgus monkey donors were mixed with MSLN expressing cancer cells of interest (CaOV3 cells and OVCAR3 cells), different amounts of a MSLN targeting trispecific antigen binding protein (containing the MH6T domain, SEQ ID NO:58) were added to the mixture and incubated at 37 ℃ for 48 hours. In parallel, a cocktail of cynomolgus PBMCs and MSLN expressing cells was incubated with varying amounts of the GFP-targeted control TriTAC molecule GFP TriTAC (SEQ ID NO:59) at 37 ℃ for 48 hours, as described above. The target cancer cells used in this assay are engineered to express luciferase. After 48 hours, the remaining viable target cells were quantified using a luminescence assay.

As shown in fig. 5, it was observed that the MSLN-targeted trispecific antigen-binding protein (containing MH6T domain) was able to efficiently direct cynomolgus PBMC to kill cells expressing MSLN (i.e., Caov3 and OVCAR), whereas the control GFP TriTAC molecule was unable to direct cynomolgus PBMC to kill the cells (also shown in fig. 5). EC targeting MSLN trispecific antigen-binding protein (containing MH6T domain)₅₀Values of 2.9pM for OVCAR3 cells and 3.0pM for Caov3 cells, compared to the EC observed with human T cells₅₀The values were not significantly different as shown in table II.

Practice ofExample 8: trispecific antigen binding targeting MSLN in the presence or absence of human serum albumin Protein (containing MH6T domain) directs T cells to kill NCI-H2052 mesothelioma cells expressing MSLN

The objective of this study was to assess whether binding of a MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain; SEQ ID NO:58) to Human Serum Albumin (HSA) affected the ability of this protein to direct T cells to kill MSLN-expressing cells. The NCI-H2052 mesothelioma cells used in this study were engineered to express luciferase. T cells from healthy donors and MSLN-expressing cells (NCI-H2052) were pooled and different amounts of a trispecific antigen-binding protein targeting MSLN (containing the MH6T domain) were added to the pool. The mixture was incubated at 37 ℃ for 48 hours in the presence or absence of HSA. Mixtures of NCI-H2052 cells and T cells were also incubated with a control GFP-targeting trispecific molecule GFP TRITAC (SEQ ID NO:59) in the presence or absence of HSA for 48 hours at 37 ℃. After 48 hours, the remaining viable target cells were quantified using a luminescence assay.

It was observed that a trispecific antigen-binding protein targeting MSLN (containing MH6T domain) was able to efficiently direct T cells to kill NCI-H2052 cells in the presence or absence of HSA (as shown in figure 6), whereas the control GFP TriTAC molecule was not (as also shown in figure 6). EC for cell killing in the presence of HSA was also observed₅₀The value increased by about 3.2 times (as shown in Table IV)

Further TDCC assays using MSLN-targeting trispecific antigen-binding proteins (containing the MH6T domain) with additional MSLN-expressing cell lines in the presence or absence of 15mg/ml HSA, the EC of which₅₀The values are presented in table IV.

Table IV: trispecific antigen-binding proteins targeting MSLN (containing MH6T domain) direct T cells to kill EC of MSLN-expressing cancer cells in the presence or absence of HSA₅₀Value of

Example 9: trispecific antigen binding proteins (containing MH6T domain) targeting MSLN and expression of MSLN In the presence of Caov4 cells, T cells from 4 different donors secreted TNF- α

The target cancer cell, CaOv4, used in this assay was engineered to express luciferase. In this assay, T cells from 4 different healthy donors (donor 02, donor 86, donor 35, and donor 81) and Caov4 cells were mixed together, different amounts of MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain; SEQ ID NO:58) were added, and the mixture was incubated at 37 ℃ for 48 hours. Caov4 cells and T cells were also incubated with a control trispecific molecule GFP TriTAC targeting GFP (SEQ ID NO:59) at 37 ℃ for 48 hours. Conditioned media from the TDCC assay was collected at 48 hours, after which the viability of the target cancer cells was measured using a luminescence assay. The concentration of TNF-. alpha.in conditioned media was determined using the AlphaLISA assay kit (Perkin Elmer).

It was observed that TNF- α was secreted into the culture medium in the presence of the Caov4 cells and the MSLN-targeting trispecific antigen-binding protein (containing MH6T domain), but not in the presence of the Caov4 cells and the control GFP TriTAC molecule, as shown in fig. 7.

Furthermore, in the presence of a trispecific antigen-binding protein targeting MSLN (containing MH6T domain), effective killing was observed for T cells from all 4 healthy donors, but not in the presence of the control GFP TriTAC molecule. TDCC assays were also established for other MSLN expressing cell lines (Caov3 cells, OVCAR3 cells, and OVCAR8 cells) and similar TNF- α expression was observed. MSLN-targeted trispecific antigen-binding proteins (containing MH6T domain) induce TNF-alpha expression of EC₅₀The values are presented in table V. However, no trispecific antigen-binding protein targeting MSLN (containing the MH6T domain) was observed to direct TNF- α secretion when assayed using cancer cells that do not express MSLN (NCI-H510A cells or MDAPCa2b cells) (data not shown). Therefore, the temperature of the molten metal is controlled,this study shows that a trispecific antigen-binding protein (containing MH6T domain) targeting MSLN is able to activate T cells in the presence of target cancer cells expressing MSLN.

Table V: trispecific antigen-binding proteins targeting MSLN (containing MH6T domain) induced TNF-alpha expressing EC from T cells from 4 different T cell donors and 4 different MSLN expressing cell lines₅₀Value of

Example 10: trispecific antigen binding proteins (containing MH6T domain) targeting MSLN and expression of MSLN Activation of CD69 expression on T cells from 4 different donors in the presence of OVCAR8 cells

OVCAR8 cells used in this assay were engineered to express luciferase. In this experiment, T cells from 4 different healthy donors (donor 02, donor 86, donor 35 and donor 81) and OVCAR8 cells were mixed together, different amounts of MSLN-targeting trispecific antigen-binding protein (containing MH6T domain; SEQ ID NO:58) were added, and the mixture was incubated at 37 ℃ for 48 hours. OVCAR8 cells and T cells were also incubated with a GFP-targeting control trispecific molecule GFP TriTAC (SEQ ID NO:59) for 48 hours at 37 ℃. After 48 hours, T cells were harvested and CD69 expression on T cells was measured by flow cytometry.

As shown in figure 8, CD69 expression was detected on T cells from all 4 healthy donors in the presence of OVCAR8 cells and a MSLN-targeting trispecific antigen-binding protein (containing MH6T domain), but no CD69 expression was detected in the presence of negative controls GFP TriTAC and OVCAR8 cells. TDCC assays were also established for other MSLN expressing cells (Caov3 cells, OVCAR3 cells, and OVCAR8 cells) and similar CD69 expression was observed. In Table VI presentMSLN-targeting trispecific antigen-binding proteins (containing MH6T domain) induce EC activation of CD69 in Caov3 cells and OVCAR8 cells₅₀The value is obtained.

Table VI: CD69 expression activated EC50 values on T cells from 4 different donors in the presence of a trispecific antigen-binding protein targeting MSLN (containing MH6T domain) and OVCAR8 or Caov3 cells expressing MSLN.

When this experiment was performed using cancer cells that do not express MSLN (NCI-H510A cells or MDAPCa2b cells), no CD69 activation induced by the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) was observed (data not shown). Thus, this study demonstrates that a trispecific antigen-binding protein (containing the MH6T domain) targeting MSLN is able to activate T cells in the presence of target cancer cells expressing MSLN.

Example 11: trispecific antigen-binding proteins targeting MSLN (containing MH6T domain) and expressing MSLN ≥ s Measurement of binding of MSLN-null cell lines

For this study, certain MSLN-expressing cancer cells of interest (Caov3 cells, CaOV4 cells, OVCAR3 cells, and OVCAR8 cells) and certain MSLN-non-expressing cancer cells (MDAPCa2b cells and NCI-H510A cells) were incubated with a MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain; SEQ ID NO:58) or a control GFP TRITAC molecule (SEQ ID NO: 59). After incubation, cells were washed to remove unbound MH6T or GFP TriTAC molecules and further incubated with a second antibody conjugated to Alexa Fluor 647, which was able to recognize the anti-albumin domain in the TriTAC molecule. The binding of a trispecific antigen-binding protein (containing MH6T domain) targeting MSLN or GFP TriTAC to cells expressing MSLN or not expressing MSLN was measured by flow cytometry.

Firm binding of MSLN-targeting trispecific antigen-binding proteins (containing the MH6T domain) to MSLN-expressing cell lines (Caov3, Caov4, OVCAR3, and OVCAR8) was observed as shown in FIG. 9A (top left panel shows binding of MSLN-targeting trispecific target antigen-binding proteins containing the MH6T domain to Caov3 cells; top right panel shows binding of MSLN-targeting trispecific target antigen-binding proteins containing the MH6T domain to Caov4 cells; bottom left panel shows binding of MSLN-targeting trispecific target antigen-binding proteins containing the MH6T domain to OV3 cells; bottom right panel shows binding of MSLN-targeting trispecific target antigen-binding proteins containing the MH6T domain to OVCAR8 cells); as shown in fig. 9B, no binding was observed in cell lines that did not express MSLN (left panel shows that the MSLN-targeting trispecific antigen-binding protein (containing MH6T domain) failed to bind to MDAPCa2B cells, right panel shows that the MSLN-targeting trispecific antigen-binding protein (containing MH6T domain) failed to bind to NCI-H510A cells). Furthermore, no binding was observed when any cell type was incubated with GFP TriTAC molecules, as shown in fig. 9A and 9B.

Example 12: trispecific antigen-binding proteins targeting MSLN (containing MH6T domain) with donor-derived antigen binding proteins Measurement of binding of T cells

For this study, T cells from 4 healthy donors were incubated with MSLN-targeting trispecific antigen-binding protein (containing MH6T domain; SEQ ID NO:58) or buffer (as negative control). After incubation, cells were washed to remove unbound trispecific antigen-binding protein targeting MSLN (containing MH6T domain) and further incubated with Alexa Fluor 647 conjugated secondary antibody capable of recognizing the anti-albumin domain in the trispecific antigen-binding protein targeting MSLN (containing MH6T domain). Binding was measured by flow cytometry.

Firm binding was observed in T cells from all four donors treated with a trispecific antigen-binding protein targeting MSLN (containing the MH6T domain) as shown in figure 10 (top left panel showing binding of the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) to T cells from donor 2; top right panel showing binding of the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) to T cells from donor 35; bottom left panel showing binding of the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) to T cells from donor 41; bottom right panel showing binding of the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) to T cells from donor 81).

Example 13: mice treated with a trispecific antigen-binding protein targeting MSLN (containing MH6T domain) Inhibition of tumor growth in humans

For this study, 10 will be⁷NCI-H292 cells and 10⁷Personal PBMCs were co-implanted subcutaneously into two groups of NCG mice (8 mice per group). After 5 days, a group of mice was injected daily with a trispecific antigen-binding protein targeting MSLN (containing MH6T domain; SEQ ID NO:58) at a dose of 0.25mg/kg for 10 days (days 5-14); while another group of mice was injected with vehicle control. Tumor volumes were measured every few days and the study was terminated on day 36. Significant tumor growth inhibition was observed in mice injected with the trispecific antigen-binding protein targeting MSLN (containing MH6T domain) compared to vehicle control injected mice, as shown in figure 11.

Example 12: trispecific antigen binding proteins targeting MSLN (containing MH6T domain) in cynomolgus monkeys Pharmacokinetics

For this study, two cynomolgus monkeys were injected intravenously with a 10mg/kg dose of MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain; SEQ ID NO:58) and serum samples were collected at different time points after injection. The amount of MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain) in serum was measured in an electrochemiluminescence assay using anti-idiotypic antibodies that recognize the MSLN-targeting trispecific antigen-binding protein (containing the MH6T domain). Figure 12 shows a graph of serum levels of a trispecific antigen-binding protein (containing MH6T domain) targeting MSLN at different time points. These data were then used to calculate the pharmacokinetic properties of a trispecific antigen-binding protein targeting MSLN (containing the MH6T domain), as shown in table VII.

Table VII: pharmacokinetic parameters of MSLN-targeting trispecific antigen-binding proteins (containing MH6T domain)

Sequence listing

CDR1 sequences of various exemplary MSLN binding domains of the present disclosure

CDR2 sequences of various exemplary MSLN binding domains of the present disclosure

CDR3 sequences of various exemplary MSLN binding domains of the present disclosure

Framework region 1(f1) sequences of various exemplary MSLN binding domains

Framework region 2(f2) sequences of various exemplary MSLN binding domains

Framework region 3(f3) sequences of various exemplary MSLN binding domains

Framework region 4(f4) sequences of various exemplary MSLN binding domains

Sequence listing

<110> Harpoon therapeutic Co

<120> mesothelin binding protein

<130> 47517-719.7111

<140>

<141>

<150> 62/657,417

<151> 2018-04-13

<150> 62/505,719

<151> 2017-05-12

<160> 336

<170> PatentIn version 3.5

<210> 1

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 1

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Val Arg

20 25 30

Gly Met Ala Trp Tyr Arg Gln Ala Gly Asn Asn Arg Ala Leu Val Ala

35 40 45

Thr Met Asn Pro Asp Gly Phe Pro Asn Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Trp Asp Ile Ala Glu Asn Thr Val Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Asn Ser Glu Asp Thr Thr Val Tyr Tyr Cys Asn Ser

85 90 95

Gly Pro Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

100 105 110

<210> 2

<211> 123

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 2

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Pro Ser Ile Glu

20 25 30

Gln Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ala Leu Thr Ser Gly Gly Arg Ala Asn Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Gly Asp Asn Val Arg Asn Met Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser

85 90 95

Ala Gly Arg Phe Lys Gly Asp Tyr Ala Gln Arg Ser Gly Met Asp Tyr

100 105 110

Trp Gly Lys Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 3

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 3

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Phe Ser Gly Thr Thr Tyr Thr Phe Asp

20 25 30

Leu Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Thr Val Val

35 40 45

Ala Ser Ile Ser Ser Asp Gly Arg Thr Ser Tyr Ala Asp Ser Val Arg

50 55 60

Gly Arg Phe Thr Ile Ser Gly Glu Asn Gly Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Leu Glu Asp Thr Ala Val Tyr Tyr Cys Leu

85 90 95

Gly Gln Arg Ser Gly Val Arg Ala Phe Trp Gly Gln Gly Thr Gln Val

100 105 110

Thr Val Ser Ser

115

<210> 4

<211> 123

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 4

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Ser Thr Ser Asn Ile Asn

20 25 30

Asn Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Val Ile Thr Arg Gly Gly Tyr Ala Ile Tyr Leu Asp Ala Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Asn Asn Ala Ile Tyr Leu

65 70 75 80

Glu Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Val Cys Asn

85 90 95

Ala Asp Arg Val Glu Gly Thr Ser Gly Gly Pro Gln Leu Arg Asp Tyr

100 105 110

Phe Gly Gln Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 5

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 5

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Gly Ile Asn

20 25 30

Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Val Ile Ser Arg Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Glu Asn Thr Val Ser Leu

65 70 75 80

Gln Met Asn Thr Leu Lys Pro Glu Asp Thr Ala Val Tyr Phe Cys Asn

85 90 95

Ala Arg Thr Tyr Thr Arg His Asp Tyr Trp Gly Gln Gly Thr Gln Val

100 105 110

Thr Val Ser Ser

115

<210> 6

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 6

Gln Val Arg Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Ile Ser Ala Phe Arg Leu Met

20 25 30

Ser Val Arg Trp Tyr Arg Gln Asp Pro Ser Lys Gln Arg Glu Trp Val

35 40 45

Ala Thr Ile Asp Gln Leu Gly Arg Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Ala Ile Ser Lys Asp Ser Thr Arg Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Met Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Ala Gly Gly Gly Pro Leu Gly Ser Arg Trp Leu Arg Gly Arg His Trp

100 105 110

Gly Gln Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 7

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 7

Gln Val Arg Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Glu

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Pro Phe Ser Ile Asn

20 25 30

Thr Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ser Ile Ser Ser Ser Gly Asp Phe Thr Tyr Thr Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Ala Arg Arg Thr Tyr Leu Pro Arg Arg Phe Gly Ser Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 8

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 8

Gln Val Gln Pro Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Val Ser Gly Ser Asp Phe Thr Glu Asp

20 25 30

Ala Met Ala Trp Tyr Arg Gln Ala Ser Gly Lys Glu Arg Glu Ser Val

35 40 45

Ala Phe Val Ser Lys Asp Gly Lys Arg Ile Leu Tyr Leu Asp Ser Val

50 55 60

Arg Gly Arg Phe Thr Ile Ser Arg Asp Ile Asp Lys Lys Thr Val Tyr

65 70 75 80

Leu Gln Met Asp Asn Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys

85 90 95

Asn Ser Ala Pro Gly Ala Ala Arg Asn Tyr Trp Gly Gln Gly Thr Gln

100 105 110

Val Thr Val Ser Ser

115

<210> 9

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 9

Gln Val Gln Pro Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Val Ser Gly Ser Asp Phe Thr Glu Asp

20 25 30

Ala Met Ala Trp Tyr Arg Gln Ala Ser Gly Lys Glu Arg Glu Ser Val

35 40 45

Ala Phe Val Ser Lys Asp Gly Lys Arg Ile Leu Tyr Leu Asp Ser Val

50 55 60

Arg Gly Arg Phe Thr Ile Ser Arg Asp Ile Tyr Lys Lys Thr Val Tyr

65 70 75 80

Leu Gln Met Asp Asn Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys

85 90 95

Asn Ser Ala Pro Gly Ala Ala Arg Asn Val Trp Gly Gln Gly Thr Gln

100 105 110

Val Thr Val Ser Ser

115

<210> 10

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 10

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Ile Gly Gly Ser Leu Ser Arg Ser Ser Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 11

<211> 124

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 11

Gln Val Gln Ile Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Leu Thr Tyr Ser Ile Val

20 25 30

Ala Val Gly Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Met Val

35 40 45

Ala Asp Ile Ser Pro Val Gly Asn Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Lys Glu Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys His

85 90 95

Ile Val Arg Gly Trp Leu Asp Glu Arg Pro Gly Pro Gly Pro Ile Val

100 105 110

Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 12

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 12

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Gly Val Tyr

20 25 30

Gly Met Glu Trp Phe Arg Gln Ala Pro Gly Lys Gln Arg Glu Trp Val

35 40 45

Ala Ser His Thr Ser Thr Gly Tyr Val Tyr Tyr Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Ser Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Lys

85 90 95

Ala Asn Arg Gly Ser Tyr Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr

100 105 110

Val Ser Ser

115

<210> 13

<211> 114

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 13

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Thr Thr Ser Ser Ile Asn Ser

20 25 30

Met Ser Trp Tyr Arg Gln Ala Gln Gly Lys Gln Arg Glu Pro Val Ala

35 40 45

Val Ile Thr Asp Arg Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

85 90 95

Ile Ala Asp Trp Arg Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val

100 105 110

Ser Ser

<210> 14

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 14

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Leu Ser Arg Tyr

20 25 30

Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Phe Val

35 40 45

Ala Ala Ile Ser Arg Ser Gly Gly Thr Thr Arg Tyr Ser Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Ala Asn Thr Phe Tyr

65 70 75 80

Leu Gln Met Asn Asn Leu Arg Pro Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Asn Val Arg Arg Arg Gly Trp Gly Arg Thr Leu Glu Tyr Trp Gly Gln

100 105 110

Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 15

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 15

Gln Val Gln Leu Gly Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Pro Asn

20 25 30

Ala Met Ile Trp His Arg Gln Ala Pro Gly Lys Gln Arg Glu Pro Val

35 40 45

Ala Ser Ile Asn Ser Ser Gly Ser Thr Asn Tyr Gly Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Val Ser Arg Asp Ile Val Lys Asn Thr Met Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Tyr Ser Asp Phe Arg Arg Gly Thr Gln Tyr Trp Gly Gln Gly Thr Gln

100 105 110

Val Thr Val Ser Ser

115

<210> 16

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 16

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Pro Ser Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ala Thr Ser Ala Ile Thr

20 25 30

Asn Leu Gly Trp Tyr Arg Arg Ala Pro Gly Gln Val Arg Glu Met Val

35 40 45

Ala Arg Ile Ser Val Arg Glu Asp Lys Glu Asp Tyr Glu Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Gln Asn Leu Val Tyr

65 70 75 80

Leu Gln Met Asn Asn Leu Gln Pro His Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Gly Ala Gln Arg Trp Gly Arg Gly Pro Gly Thr Thr Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 17

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 17

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Arg Ile Arg

20 25 30

Val Met Arg Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val

35 40 45

Ala Val Ile Ser Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Asp Ser Gly Ile Ala Arg Asp Tyr Trp Gly Gln Gly Thr Gln Val

100 105 110

Thr Val Ser Ser

115

<210> 18

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 18

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Glu

1 5 10 15

Ser Arg Arg Leu Ser Cys Ala Val Ser Gly Asp Thr Ser Lys Phe Lys

20 25 30

Ala Val Gly Trp Tyr Arg Gln Ala Pro Gly Ala Gln Arg Glu Leu Leu

35 40 45

Ala Trp Ile Asn Asn Ser Gly Val Gly Asn Thr Ala Glu Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Arg Leu Thr Pro Glu Asp Thr Asp Val Tyr Tyr Cys Arg

85 90 95

Phe Tyr Arg Arg Phe Gly Ile Asn Lys Asn Tyr Trp Gly Gln Gly Thr

100 105 110

Gln Val Thr Val Ser Ser

115

<210> 19

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 19

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Gly Asn Lys

20 25 30

Pro Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Val Ile Ser Ser Asp Gly Gly Ser Thr Arg Tyr Ala Ala Leu Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Glu Ser Leu Val Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Asn Ala Leu Arg Thr Tyr Tyr Leu Asn Asp Pro Val Val Phe Ser Trp

100 105 110

Gly Gln Gly Thr Gln Val Thr Val Ser Ser

115 120

<210> 20

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 20

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Ser Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Leu Arg Gly Thr Leu His Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 21

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 21

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Arg Thr Asp Arg Ile Thr

20 25 30

Thr Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Thr Ile Ser Asn Arg Gly Thr Ser Asn Tyr Ala Asn Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Ala Arg Lys Trp Gly Arg Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr

100 105 110

Val Ser Ser

115

<210> 22

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 22

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Arg Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Arg Thr Ile Gly Ile Asn

20 25 30

Asp Met Ala Trp Tyr Arg Gln Ala Pro Gly Asn Gln Arg Glu Leu Val

35 40 45

Ala Thr Ile Thr Lys Gly Gly Thr Thr Asp Tyr Ala Asp Ser Val Asp

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Lys Arg Arg Glu Trp Ala Lys Asp Phe Glu Tyr Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 23

<211> 114

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 23

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Ala Ile Gly Ser Ile Asn Ser

20 25 30

Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Pro Val Ala

35 40 45

Val Ile Thr Asp Arg Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

85 90 95

Ile Ala Asp Trp Arg Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val

100 105 110

Ser Ser

<210> 24

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 24

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Leu Val

35 40 45

Ala Thr Ile Asn Arg Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Ser Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu

65 70 75 80

Gln Met Asn Arg Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 25

<211> 114

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 25

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Thr Ser Thr Thr Phe Ser Ile Asn Ser

20 25 30

Met Ser Trp Tyr Arg Gln Ala Pro Gly Asn Gln Arg Glu Pro Val Ala

35 40 45

Val Ile Thr Asn Arg Gly Thr Thr Ser Tyr Ala Asp Ser Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Val Tyr Leu Gln

65 70 75 80

Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

85 90 95

Ile Ala Asp Trp Arg Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val

100 105 110

Ser Ser

<210> 26

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 26

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Gln Val

100 105 110

Thr Val Ser Ser

115

<210> 27

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 27

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Arg Thr Ser Thr Ile Asp

20 25 30

Thr Met Tyr Trp His Arg Gln Ala Pro Gly Asn Glu Arg Glu Leu Val

35 40 45

Ala Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ala Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Val Arg Thr Thr Ser Tyr Pro Val Asp Phe Trp Gly Gln Gly Thr Gln

100 105 110

Val Thr Val Ser Ser

115

<210> 28

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 28

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Ser Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe Trp Gly Gln Gly

100 105 110

Thr Gln Val Thr Val Ser Ser

115

<210> 29

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 29

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Gly Asp Trp Ser Ala Asn

20 25 30

Phe Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Ser Gly Arg Gly Val Val Asp Tyr Val Glu Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Val Ala Ser Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

100 105 110

<210> 30

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 30

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Gly Asp Trp Ser Ala Asn

20 25 30

Phe Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Ser Gly Arg Gly Val Val Asp Tyr Val Glu Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Val Ala Ser Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

100 105 110

<210> 31

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 31

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Gly Asp Trp Ser Ala Asn

20 25 30

Phe Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Arg Ile Ser Gly Arg Gly Val Val Asp Tyr Val Glu Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Val Ala Ser Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

100 105 110

<210> 32

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 32

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 33

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 33

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 34

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 34

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ser Ser Ile Asn

20 25 30

Thr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Thr Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 35

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 35

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 36

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 36

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Gly Gly

115

<210> 37

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 37

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Gly Gly

115

<210> 38

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 38

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Arg Thr Ser Thr Ile Asp

20 25 30

Thr Met Tyr Trp His Arg Gln Ala Pro Gly Asn Glu Arg Glu Leu Val

35 40 45

Ala Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Val Arg Thr Thr Ser Tyr Pro Val Asp Phe Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Gly Gly

115

<210> 39

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 39

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Ser Thr Ile Asp

20 25 30

Thr Met Tyr Trp His Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Val Arg Thr Thr Ser Tyr Pro Val Asp Phe Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 40

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 40

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Ser Thr Ile Asp

20 25 30

Thr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser

85 90 95

Val Arg Thr Thr Ser Tyr Pro Val Asp Phe Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 41

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 41

Glu Ser Gly Gly Gly Leu Val

1 5

<210> 42

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 42

Leu Ser Cys

<210> 43

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 43

Gly Arg Phe

<210> 44

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 44

Val Thr Val Ser Ser

1 5

<210> 45

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 45

Gln Leu Val Glu Ser Gly Gly Gly

1 5

<210> 46

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 46

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

1 5 10

<210> 47

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 47

Ala Ser Gly

<210> 48

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 48

Arg Gln Ala Pro Gly

1 5

<210> 49

<211> 33

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 49

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu

1 5 10 15

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

20 25 30

Cys

<210> 50

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 50

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 51

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 51

Gly Arg Thr Phe Ser Val Arg Gly Met Ala

1 5 10

<210> 52

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 52

Ile Asn Ser Ser Gly Ser Thr Asn Tyr Gly

1 5 10

<210> 53

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 53

Asn Ala Gly Gly Gly Pro Leu Gly Ser Arg

1 5 10

<210> 54

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 54

Gly Gly Asp Trp Ser Ala Asn Phe Met Tyr

1 5 10

<210> 55

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 55

Ile Ser Ser Gly Gly Ser Thr Asn Val Arg

1 5 10

<210> 56

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 56

Asn Ala Asp Thr Ile Gly Thr Ala Arg Asp

1 5 10

<210> 57

<211> 630

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 57

Met Ala Leu Pro Thr Ala Arg Pro Leu Leu Gly Ser Cys Gly Thr Pro

1 5 10 15

Ala Leu Gly Ser Leu Leu Phe Leu Leu Phe Ser Leu Gly Trp Val Gln

20 25 30

Pro Ser Arg Thr Leu Ala Gly Glu Thr Gly Gln Glu Ala Ala Pro Leu

35 40 45

Asp Gly Val Leu Ala Asn Pro Pro Asn Ile Ser Ser Leu Ser Pro Arg

50 55 60

Gln Leu Leu Gly Phe Pro Cys Ala Glu Val Ser Gly Leu Ser Thr Glu

65 70 75 80

Arg Val Arg Glu Leu Ala Val Ala Leu Ala Gln Lys Asn Val Lys Leu

85 90 95

Ser Thr Glu Gln Leu Arg Cys Leu Ala His Arg Leu Ser Glu Pro Pro

100 105 110

Glu Asp Leu Asp Ala Leu Pro Leu Asp Leu Leu Leu Phe Leu Asn Pro

115 120 125

Asp Ala Phe Ser Gly Pro Gln Ala Cys Thr Arg Phe Phe Ser Arg Ile

130 135 140

Thr Lys Ala Asn Val Asp Leu Leu Pro Arg Gly Ala Pro Glu Arg Gln

145 150 155 160

Arg Leu Leu Pro Ala Ala Leu Ala Cys Trp Gly Val Arg Gly Ser Leu

165 170 175

Leu Ser Glu Ala Asp Val Arg Ala Leu Gly Gly Leu Ala Cys Asp Leu

180 185 190

Pro Gly Arg Phe Val Ala Glu Ser Ala Glu Val Leu Leu Pro Arg Leu

195 200 205

Val Ser Cys Pro Gly Pro Leu Asp Gln Asp Gln Gln Glu Ala Ala Arg

210 215 220

Ala Ala Leu Gln Gly Gly Gly Pro Pro Tyr Gly Pro Pro Ser Thr Trp

225 230 235 240

Ser Val Ser Thr Met Asp Ala Leu Arg Gly Leu Leu Pro Val Leu Gly

245 250 255

Gln Pro Ile Ile Arg Ser Ile Pro Gln Gly Ile Val Ala Ala Trp Arg

260 265 270

Gln Arg Ser Ser Arg Asp Pro Ser Trp Arg Gln Pro Glu Arg Thr Ile

275 280 285

Leu Arg Pro Arg Phe Arg Arg Glu Val Glu Lys Thr Ala Cys Pro Ser

290 295 300

Gly Lys Lys Ala Arg Glu Ile Asp Glu Ser Leu Ile Phe Tyr Lys Lys

305 310 315 320

Trp Glu Leu Glu Ala Cys Val Asp Ala Ala Leu Leu Ala Thr Gln Met

325 330 335

Asp Arg Val Asn Ala Ile Pro Phe Thr Tyr Glu Gln Leu Asp Val Leu

340 345 350

Lys His Lys Leu Asp Glu Leu Tyr Pro Gln Gly Tyr Pro Glu Ser Val

355 360 365

Ile Gln His Leu Gly Tyr Leu Phe Leu Lys Met Ser Pro Glu Asp Ile

370 375 380

Arg Lys Trp Asn Val Thr Ser Leu Glu Thr Leu Lys Ala Leu Leu Glu

385 390 395 400

Val Asn Lys Gly His Glu Met Ser Pro Gln Ala Pro Arg Arg Pro Leu

405 410 415

Pro Gln Val Ala Thr Leu Ile Asp Arg Phe Val Lys Gly Arg Gly Gln

420 425 430

Leu Asp Lys Asp Thr Leu Asp Thr Leu Thr Ala Phe Tyr Pro Gly Tyr

435 440 445

Leu Cys Ser Leu Ser Pro Glu Glu Leu Ser Ser Val Pro Pro Ser Ser

450 455 460

Ile Trp Ala Val Arg Pro Gln Asp Leu Asp Thr Cys Asp Pro Arg Gln

465 470 475 480

Leu Asp Val Leu Tyr Pro Lys Ala Arg Leu Ala Phe Gln Asn Met Asn

485 490 495

Gly Ser Glu Tyr Phe Val Lys Ile Gln Ser Phe Leu Gly Gly Ala Pro

500 505 510

Thr Glu Asp Leu Lys Ala Leu Ser Gln Gln Asn Val Ser Met Asp Leu

515 520 525

Ala Thr Phe Met Lys Leu Arg Thr Asp Ala Val Leu Pro Leu Thr Val

530 535 540

Ala Glu Val Gln Lys Leu Leu Gly Pro His Val Glu Gly Leu Lys Ala

545 550 555 560

Glu Glu Arg His Arg Pro Val Arg Asp Trp Ile Leu Arg Gln Arg Gln

565 570 575

Asp Asp Leu Asp Thr Leu Gly Leu Gly Leu Gln Gly Gly Ile Pro Asn

580 585 590

Gly Tyr Leu Val Leu Asp Leu Ser Met Gln Glu Ala Leu Ser Gly Thr

595 600 605

Pro Cys Leu Leu Gly Pro Gly Pro Val Leu Thr Val Leu Ala Leu Leu

610 615 620

Leu Ala Ser Thr Leu Ala

625 630

<210> 58

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 58

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 59

<211> 503

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 59

Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr

20 25 30

Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val

35 40 45

Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr

100 105 110

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu

115 120 125

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu

130 135 140

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp

145 150 155 160

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser

165 170 175

Gly Ser Gly Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe

180 185 190

Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn

195 200 205

Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly

210 215 220

Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly

225 230 235 240

Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

245 250 255

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala

260 265 270

Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala

275 280 285

Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn

290 295 300

Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile

305 310 315 320

Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu

325 330 335

Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe

340 345 350

Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu

355 360 365

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

370 375 380

Gly Gly Gly Ser Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val

385 390 395 400

Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala

405 410 415

Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln

420 425 430

Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr

435 440 445

Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr

450 455 460

Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu

465 470 475 480

Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val

485 490 495

Leu His His His His His His

500

<210> 60

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 60

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 61

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 61

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val

35 40 45

Ala Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 62

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Polypeptides

<400> 62

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ile Arg

20 25 30

Ala Met Arg Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

85 90 95

Asp Thr Ile Gly Thr Ala Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 63

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 63

Gly Arg Thr Phe Ser Val Arg Gly Met Ala

1 5 10

<210> 64

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 64

Gly Ser Ile Pro Ser Ile Glu Gln Met Gly

1 5 10

<210> 65

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 65

Gly Thr Thr Tyr Thr Phe Asp Leu Met Ser

1 5 10

<210> 66

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 66

Gly Ser Thr Ser Asn Ile Asn Asn Met Arg

1 5 10

<210> 67

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 67

Gly Ser Thr Phe Gly Ile Asn Ala Met Gly

1 5 10

<210> 68

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 68

Ile Ser Ala Phe Arg Leu Met Ser Val Arg

1 5 10

<210> 69

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 69

Gly Arg Pro Phe Ser Ile Asn Thr Met Gly

1 5 10

<210> 70

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 70

Gly Ser Asp Phe Thr Glu Asp Ala Met Ala

1 5 10

<210> 71

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 71

Gly Ser Asp Phe Thr Glu Asp Ala Met Ala

1 5 10

<210> 72

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 72

Gly Phe Thr Phe Ser Ser Phe Gly Met Ser

1 5 10

<210> 73

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 73

Gly Leu Thr Tyr Ser Ile Val Ala Val Gly

1 5 10

<210> 74

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 74

Gly Leu Thr Phe Gly Val Tyr Gly Met Glu

1 5 10

<210> 75

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 75

Thr Thr Ser Ser Ile Asn Ser Met Ser

1 5

<210> 76

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 76

Gly Arg Thr Leu Ser Arg Tyr Ala Met Gly

1 5 10

<210> 77

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 77

Gly Ser Ile Phe Ser Pro Asn Ala Met Ile

1 5 10

<210> 78

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 78

Gly Ala Thr Ser Ala Ile Thr Asn Leu Gly

1 5 10

<210> 79

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 79

Gly Ser Thr Phe Arg Ile Arg Val Met Arg

1 5 10

<210> 80

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 80

Gly Asp Thr Ser Lys Phe Lys Ala Val Gly

1 5 10

<210> 81

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 81

Gly Ser Thr Phe Gly Asn Lys Pro Met Gly

1 5 10

<210> 82

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 82

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 83

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 83

Gly Arg Thr Asp Arg Ile Thr Thr Met Gly

1 5 10

<210> 84

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 84

Gly Arg Thr Ile Gly Ile Asn Asp Met Ala

1 5 10

<210> 85

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 85

Ala Ile Gly Ser Ile Asn Ser Met Ser

1 5

<210> 86

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 86

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 87

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 87

Thr Thr Phe Ser Ile Asn Ser Met Ser

1 5

<210> 88

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 88

Gly Ser Thr Phe Ser Ile Arg Ala Met Arg

1 5 10

<210> 89

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 89

Gly Arg Thr Ser Thr Ile Asp Thr Met Tyr

1 5 10

<210> 90

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 90

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 91

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 91

Gly Gly Asp Trp Ser Ala Asn Phe Met Tyr

1 5 10

<210> 92

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 92

Gly Gly Asp Trp Ser Ala Asn Phe Met Tyr

1 5 10

<210> 93

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 93

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 94

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 94

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 95

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 95

Gly Ser Thr Ser Ser Ile Asn Thr Met Tyr

1 5 10

<210> 96

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 96

Gly Ser Thr Phe Ser Ile Arg Ala Met Arg

1 5 10

<210> 97

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 97

Gly Ser Thr Phe Ser Ile Arg Ala Met Arg

1 5 10

<210> 98

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 98

Gly Ser Thr Phe Ser Ile Arg Ala Met Arg

1 5 10

<210> 99

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 99

Gly Arg Thr Ser Thr Ile Asp Thr Met Tyr

1 5 10

<210> 100

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 100

Gly Arg Thr Ser Thr Ile Asp Thr Met Tyr

1 5 10

<210> 101

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 101

Gly Arg Thr Ser Thr Ile Asp Thr Met Tyr

1 5 10

<210> 102

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 102

Thr Met Asn Pro Asp Gly Phe Pro Asn Tyr Ala Asp Ala Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 103

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 103

Ala Leu Thr Ser Gly Gly Arg Ala Asn Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 104

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 104

Ser Ile Ser Ser Asp Gly Arg Thr Ser Tyr Ala Asp Ser Val Arg Gly

1 5 10 15

Arg Phe Thr

<210> 105

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 105

Val Ile Thr Arg Gly Gly Tyr Ala Ile Tyr Leu Asp Ala Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 106

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 106

Val Ile Ser Arg Gly Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 107

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 107

Thr Ile Asp Gln Leu Gly Arg Thr Asn Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Ala

<210> 108

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 108

Ser Ile Ser Ser Ser Gly Asp Phe Thr Tyr Thr Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 109

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 109

Phe Val Ser Lys Asp Gly Lys Arg Ile Leu Tyr Leu Asp Ser Val Arg

1 5 10 15

Gly Arg Phe Thr

<210> 110

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 110

Phe Val Ser Lys Asp Gly Lys Arg Ile Leu Tyr Leu Asp Ser Val Arg

1 5 10 15

Gly Arg Phe Thr

<210> 111

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 111

Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly Arg Phe Thr

<210> 112

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 112

Asp Ile Ser Pro Val Gly Asn Thr Asn Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 113

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 113

Ser His Thr Ser Thr Gly Tyr Val Tyr Tyr Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 114

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 114

Val Ile Thr Asp Arg Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 115

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 115

Ala Ile Ser Arg Ser Gly Gly Thr Thr Arg Tyr Ser Asp Ser Val Lys

1 5 10 15

Gly Arg Phe Thr

<210> 116

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 116

Ser Ile Asn Ser Ser Gly Ser Thr Asn Tyr Gly Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 117

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 117

Arg Ile Ser Val Arg Glu Asp Lys Glu Asp Tyr Glu Asp Ser Val Lys

1 5 10 15

Gly Arg Phe Thr

<210> 118

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 118

Val Ile Ser Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg

1 5 10 15

Phe Thr

<210> 119

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 119

Trp Ile Asn Asn Ser Gly Val Gly Asn Thr Ala Glu Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 120

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 120

Val Ile Ser Ser Asp Gly Gly Ser Thr Arg Tyr Ala Ala Leu Val Lys

1 5 10 15

Gly Arg Phe Thr

<210> 121

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 121

Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Ser

<210> 122

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 122

Thr Ile Ser Asn Arg Gly Thr Ser Asn Tyr Ala Asn Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 123

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 123

Thr Ile Thr Lys Gly Gly Thr Thr Asp Tyr Ala Asp Ser Val Asp Gly

1 5 10 15

Arg Phe Thr

<210> 124

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 124

Val Ile Thr Asp Arg Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 125

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 125

Thr Ile Asn Arg Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Ser

<210> 126

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 126

Val Ile Thr Asn Arg Gly Thr Thr Ser Tyr Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 127

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 127

Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly Arg

1 5 10 15

Phe Thr

<210> 128

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 128

Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 129

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 129

Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Ser

<210> 130

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 130

Arg Ile Ser Gly Arg Gly Val Val Asp Tyr Val Glu Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 131

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 131

Arg Ile Ser Gly Arg Gly Val Val Asp Tyr Val Glu Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 132

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 132

Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 133

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 133

Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 134

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 134

Phe Ile Ser Ser Gly Gly Ser Thr Asn Val Arg Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 135

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 135

Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly Arg

1 5 10 15

Phe Thr

<210> 136

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 136

Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly Arg

1 5 10 15

Phe Thr

<210> 137

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 137

Val Ile Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ala Val Lys Gly Arg

1 5 10 15

Phe Thr

<210> 138

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 138

Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 139

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 139

Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 140

<211> 19

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 140

Tyr Val Thr Ser Arg Gly Thr Ser Asn Val Ala Asp Ser Val Lys Gly

1 5 10 15

Arg Phe Thr

<210> 141

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 141

Gly Pro Tyr

<210> 142

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 142

Gly Arg Phe Lys Gly Asp Tyr Ala Gln Arg Ser Gly Met Asp Tyr

1 5 10 15

<210> 143

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 143

Gln Arg Ser Gly Val Arg Ala Phe

1 5

<210> 144

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 144

Asp Arg Val Glu Gly Thr Ser Gly Gly Pro Gln Leu Arg Asp Tyr

1 5 10 15

<210> 145

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 145

Arg Thr Tyr Thr Arg His Asp Tyr

1 5

<210> 146

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 146

Gly Gly Gly Pro Leu Gly Ser Arg Trp Leu Arg Gly Arg His

1 5 10

<210> 147

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 147

Arg Arg Thr Tyr Leu Pro Arg Arg Phe Gly Ser

1 5 10

<210> 148

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 148

Ala Pro Gly Ala Ala Arg Asn Tyr

1 5

<210> 149

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 149

Ala Pro Gly Ala Ala Arg Asn Val

1 5

<210> 150

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 150

Gly Gly Ser Leu Ser Arg Ser Ser

1 5

<210> 151

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 151

Val Arg Gly Trp Leu Asp Glu Arg Pro Gly Pro Gly Pro Ile Val Tyr

1 5 10 15

<210> 152

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 152

Asn Arg Gly Ser Tyr Glu Tyr

1 5

<210> 153

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 153

Ile Ala Asp Trp Arg Gly Tyr

1 5

<210> 154

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 154

Arg Arg Arg Gly Trp Gly Arg Thr Leu Glu Tyr

1 5 10

<210> 155

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 155

Ser Asp Phe Arg Arg Gly Thr Gln Tyr

1 5

<210> 156

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 156

Gln Arg Trp Gly Arg Gly Pro Gly Thr Thr

1 5 10

<210> 157

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 157

Asp Asp Ser Gly Ile Ala Arg Asp Tyr

1 5

<210> 158

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 158

Tyr Arg Arg Phe Gly Ile Asn Lys Asn Tyr

1 5 10

<210> 159

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 159

Leu Arg Thr Tyr Tyr Leu Asn Asp Pro Val Val Phe Ser

1 5 10

<210> 160

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 160

Tyr Ile Pro Leu Arg Gly Thr Leu His Asp Tyr

1 5 10

<210> 161

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 161

Arg Lys Trp Gly Arg Asn Tyr

1 5

<210> 162

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 162

Lys Arg Arg Glu Trp Ala Lys Asp Phe Glu Tyr

1 5 10

<210> 163

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 163

Ile Ala Asp Trp Arg Gly Tyr

1 5

<210> 164

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 164

Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe

1 5 10

<210> 165

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 165

Ile Ala Asp Trp Arg Gly Tyr

1 5

<210> 166

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 166

Asp Thr Ile Gly Thr Ala Arg Asp Tyr

1 5

<210> 167

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 167

Arg Thr Thr Ser Tyr Pro Val Asp Phe

1 5

<210> 168

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 168

Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe

1 5 10

<210> 169

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 169

Ala Ser Tyr

<210> 170

<211> 3

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 170

Ala Ser Tyr

<210> 171

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 171

Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe

1 5 10

<210> 172

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 172

Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe

1 5 10

<210> 173

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 173

Tyr Ile Pro Tyr Gly Gly Thr Leu His Asp Phe

1 5 10

<210> 174

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 174

Asp Thr Ile Gly Thr Ala Arg Asp Tyr

1 5

<210> 175

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 175

Asp Thr Ile Gly Thr Ala Arg Asp Tyr

1 5

<210> 176

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 176

Asp Thr Ile Gly Thr Ala Arg Asp Tyr

1 5

<210> 177

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 177

Arg Thr Thr Ser Tyr Pro Val Asp Phe

1 5

<210> 178

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 178

Arg Thr Thr Ser Tyr Pro Val Asp Phe

1 5

<210> 179

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 179

Arg Thr Thr Ser Tyr Pro Val Asp Phe

1 5

<210> 180

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 180

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 181

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 181

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 182

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 182

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Phe Ser

20 25

<210> 183

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 183

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser

20 25

<210> 184

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 184

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 185

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 185

Gln Val Arg Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 186

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 186

Gln Val Arg Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Glu

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 187

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 187

Gln Val Gln Pro Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Val Ser

20 25

<210> 188

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 188

Gln Val Gln Pro Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Val Ser

20 25

<210> 189

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 189

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 190

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 190

Gln Val Gln Ile Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser

20 25

<210> 191

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 191

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 192

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 192

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 193

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 193

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 194

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 194

Gln Val Gln Leu Gly Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 195

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 195

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Pro Ser Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 196

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 196

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 197

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 197

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Glu

1 5 10 15

Ser Arg Arg Leu Ser Cys Ala Val Ser

20 25

<210> 198

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 198

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 199

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 199

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 200

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 200

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser

20 25

<210> 201

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 201

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Arg Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser

20 25

<210> 202

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 202

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 203

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 203

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 204

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 204

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Thr Ser

20 25

<210> 205

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 205

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Thr Leu Ser Cys Ala Ala Ser

20 25

<210> 206

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 206

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser

20 25

<210> 207

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 207

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 208

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 208

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 209

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 209

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 210

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 210

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 211

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 211

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 212

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 212

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 213

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 213

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 214

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 214

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 215

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 215

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 216

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 216

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser

20 25

<210> 217

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 217

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 218

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 218

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 219

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 219

Trp Tyr Arg Gln Ala Gly Asn Asn Arg Ala Leu Val Ala

1 5 10

<210> 220

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 220

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 221

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 221

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Thr Val Val Ala

1 5 10

<210> 222

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 222

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 223

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 223

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 224

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 224

Trp Tyr Arg Gln Asp Pro Ser Lys Gln Arg Glu Trp Val Ala

1 5 10

<210> 225

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 225

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 226

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 226

Trp Tyr Arg Gln Ala Ser Gly Lys Glu Arg Glu Ser Val Ala

1 5 10

<210> 227

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 227

Trp Tyr Arg Gln Ala Ser Gly Lys Glu Arg Glu Ser Val Ala

1 5 10

<210> 228

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 228

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 229

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 229

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Met Val Ala

1 5 10

<210> 230

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 230

Trp Phe Arg Gln Ala Pro Gly Lys Gln Arg Glu Trp Val Ala

1 5 10

<210> 231

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 231

Trp Tyr Arg Gln Ala Gln Gly Lys Gln Arg Glu Pro Val Ala

1 5 10

<210> 232

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 232

Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Phe Val Ala

1 5 10

<210> 233

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 233

Trp His Arg Gln Ala Pro Gly Lys Gln Arg Glu Pro Val Ala

1 5 10

<210> 234

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 234

Trp Tyr Arg Arg Ala Pro Gly Gln Val Arg Glu Met Val Ala

1 5 10

<210> 235

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 235

Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val Ala

1 5 10

<210> 236

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 236

Trp Tyr Arg Gln Ala Pro Gly Ala Gln Arg Glu Leu Leu Ala

1 5 10

<210> 237

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 237

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 238

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 238

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 239

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 239

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 240

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 240

Trp Tyr Arg Gln Ala Pro Gly Asn Gln Arg Glu Leu Val Ala

1 5 10

<210> 241

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 241

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Pro Val Ala

1 5 10

<210> 242

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 242

Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Leu Val Ala

1 5 10

<210> 243

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 243

Trp Tyr Arg Gln Ala Pro Gly Asn Gln Arg Glu Pro Val Ala

1 5 10

<210> 244

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 244

Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val Ala

1 5 10

<210> 245

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 245

Trp His Arg Gln Ala Pro Gly Asn Glu Arg Glu Leu Val Ala

1 5 10

<210> 246

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 246

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 247

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 247

Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala

1 5 10

<210> 248

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 248

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 249

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 249

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 250

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 250

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 251

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 251

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 252

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 252

Trp Tyr Arg Gln Ala Pro Gly Thr Glu Arg Asp Leu Val Ala

1 5 10

<210> 253

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 253

Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 254

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 254

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 255

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 255

Trp His Arg Gln Ala Pro Gly Asn Glu Arg Glu Leu Val Ala

1 5 10

<210> 256

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 256

Trp His Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala

1 5 10

<210> 257

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 257

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 258

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 258

Ile Ser Trp Asp Ile Ala Glu Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Asn Ser Glu Asp Thr Thr Val Tyr Tyr Cys Asn Ser

20 25

<210> 259

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 259

Ile Ser Gly Asp Asn Val Arg Asn Met Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ser Ala

20 25

<210> 260

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 260

Ile Ser Gly Glu Asn Gly Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Leu Glu Asp Thr Ala Val Tyr Tyr Cys Leu Gly

20 25

<210> 261

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 261

Ile Ser Arg Asp Asn Ala Asn Asn Ala Ile Tyr Leu Glu Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Val Cys Asn Ala

20 25

<210> 262

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 262

Ile Ser Arg Asp Asn Ala Glu Asn Thr Val Ser Leu Gln Met Asn Thr

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Phe Cys Asn Ala

20 25

<210> 263

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 263

Ile Ser Lys Asp Ser Thr Arg Asn Thr Val Tyr Leu Gln Met Asn Met

1 5 10 15

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 264

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 264

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 265

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 265

Ile Ser Arg Asp Ile Asp Lys Lys Thr Val Tyr Leu Gln Met Asp Asn

1 5 10 15

Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ser

20 25

<210> 266

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 266

Ile Ser Arg Asp Ile Tyr Lys Lys Thr Val Tyr Leu Gln Met Asp Asn

1 5 10 15

Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ser

20 25

<210> 267

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 267

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile

20 25

<210> 268

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 268

Ile Ser Lys Glu Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys His Ile

20 25

<210> 269

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 269

Ile Ser Arg Asp Asn Ala Lys Ser Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Lys Ala

20 25

<210> 270

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 270

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

20 25

<210> 271

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 271

Ile Ser Arg Asp Asn Ala Ala Asn Thr Phe Tyr Leu Gln Met Asn Asn

1 5 10 15

Leu Arg Pro Asp Asp Thr Ala Val Tyr Tyr Cys Asn Val

20 25

<210> 272

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 272

Val Ser Arg Asp Ile Val Lys Asn Thr Met Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ser Tyr

20 25

<210> 273

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 273

Ile Ser Arg Asp Asn Thr Gln Asn Leu Val Tyr Leu Gln Met Asn Asn

1 5 10 15

Leu Gln Pro His Asp Thr Ala Ile Tyr Tyr Cys Gly Ala

20 25

<210> 274

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 274

Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Asn

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 275

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 275

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Arg

1 5 10 15

Leu Thr Pro Glu Asp Thr Asp Val Tyr Tyr Cys Arg Phe

20 25

<210> 276

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 276

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Glu Ser

1 5 10 15

Leu Val Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 277

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 277

Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 278

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 278

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 279

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 279

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 280

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 280

Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

20 25

<210> 281

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 281

Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu Gln Met Asn Arg

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 282

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 282

Ile Ser Arg Asp Asn Ala Arg Asn Thr Val Tyr Leu Gln Met Asp Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Ile Tyr Thr Cys His Val

20 25

<210> 283

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 283

Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Asn

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 284

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 284

Ile Ser Arg Asp Asn Ala Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ser Val

20 25

<210> 285

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 285

Val Ser Arg Asp Ser Ala Lys Asn Ile Val Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 286

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 286

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Val

20 25

<210> 287

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 287

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Val

20 25

<210> 288

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 288

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 289

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 289

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 290

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 290

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Thr

20 25

<210> 291

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 291

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 292

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 292

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 293

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 293

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala

20 25

<210> 294

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 294

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Val

20 25

<210> 295

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 295

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Val

20 25

<210> 296

<211> 29

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 296

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

1 5 10 15

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Val

20 25

<210> 297

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 297

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 298

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 298

Trp Gly Lys Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 299

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 299

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 300

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 300

Phe Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 301

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 301

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 302

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 302

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 303

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 303

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 304

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 304

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 305

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 305

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 306

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 306

Gln Gly Thr Leu Val Thr Val Ser Ser

1 5

<210> 307

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 307

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 308

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 308

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 309

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 309

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 310

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 310

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 311

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 311

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 312

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 312

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 313

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 313

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 314

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 314

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 315

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 315

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 316

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 316

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 317

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 317

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 318

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 318

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 319

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 319

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 320

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 320

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 321

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 321

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 322

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 322

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 323

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 323

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 324

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 324

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser

1 5 10

<210> 325

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 325

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 326

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 326

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 327

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 327

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 328

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 328

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 329

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 329

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 330

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 330

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

1 5 10

<210> 331

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 331

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

1 5 10

<210> 332

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 332

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

1 5 10

<210> 333

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 333

Trp Gly Gln Gly Thr Leu Val Thr Val Ser

1 5 10

<210> 334

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 334

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 335

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

Peptides

<400> 335

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 336

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthesis of

6XHis tag

<400> 336

His His His His His His

1 5

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