HK40047151B

HK40047151B - Anti-cd47 antigen binding protein, and application thereof

Info

Publication number: HK40047151B
Application number: HK62021036942.2A
Authority: HK
Inventors: F·格罗斯维尔德; D·德拉贝克; M(R)·范哈佩恩; R·扬森; 王路凡; 刘斌; 戎一平; 何云; 周伟
Original assignee: 和铂医药(上海)有限责任公司
Priority date: 2019-08-21
Filing date: 2020-08-20
Publication date: 2022-09-16

Description

anti-CD 47 antigen binding protein and application thereof

Cross Reference to Related Applications

This application claims priority to CN2019107757316, which is incorporated by reference in its entirety.

Technical Field

The application relates to the field of biomedicine, in particular to an anti-CD 47 antigen binding protein and application thereof.

Background

CD47, also known as integrin-associated protein (IAP), ovarian cancer antigen OA3, Rh-associated antigen and MER6, is a transmembrane protein widely expressed on the cell surface and belongs to the immunoglobulin superfamily. CD47 can interact with integrins, thrombospondin (TSP-1), and signal-regulating protein alpha (SIRP alpha). CD47 was originally identified as a tumor antigen on human ovarian cancer, and a number of studies subsequently demonstrated that CD47 is also highly expressed on tumor Cells of a large proportion of human cancers (e.g., lymphoma, leukemia, breast, colon, lung, stomach, glioblastoma, glioma, ovarian, cervical, bladder, prostate, or melanoma) (see Oldenborg PA.CD47: A Cell Surface protein Multiple Functions of hepatogenic Cells in Health and disease ISRN Hemat.2013; 2013: 614619. doi: 10.1155/2013/614619), and upon binding to SIRP α, is capable of transmitting inhibitory signals to escape phagocytosis by macrophages. Thus, agents that block the CD 47-sirpa interaction may restore phagocytic uptake by CD47+ target cells and lower the threshold for macrophage activation.

In recent years, the development of drugs aiming at the tumor escape mechanism mediated by the CD47 and SIRP alpha signal pathway is a new hot door for tumor immunotherapy. Currently, the CD47 antibodies that are clinically studied include Hu5F9-G4 of form Seven, CC-90002 of Celgene, SRF-231 of Surface Oncology, and the like. However, these antibodies also have the disadvantages of low affinity, causing hemagglutination, etc. (see patent US 9045541). Therefore, there is an urgent need to develop new therapies targeting CD47 that are safer and more effective.

Disclosure of Invention

The present application provides an anti-CD 47 antigen binding protein having one or more of the following properties: 1) capable of binding human and monkey-derived CD 47; 2) CD47 capable of specifically binding to the surface of Jurkat cells and Raji cells; 3) can inhibit the binding of CD47 and SIRPa; 4) can block the binding of CD47 on the surface of Jurkat cells to SIRPa; 5) capable of binding human red blood cells; 6) does not cause coagulation reaction; 7) can cause phagocytosis of tumor cells by macrophages; 8) can inhibit tumor growth and/or tumor cell proliferation. The application also provides the application of the antigen binding protein in preventing and treating tumors.

In one aspect, the present application provides an isolated antigen binding protein comprising an antibody light chain variable region VL comprising LCDR1, LCDR2 and LCDR3, and an antibody heavy chain variable region VH comprising HCDR1, HCDR2 and HCDR3, wherein the VL comprises SEQ ID NO: 153, and VH comprises the amino acid sequence shown in SEQ ID NO: 154.

In certain embodiments, the isolated antigen binding protein has one or more of the following properties:

1) can be 3x10^-9(ii) a KD value of M or higher binds to human and monkey-derived CD47, wherein the KD value is determined by Biacore;

2) CD47 capable of specifically binding to the surface of Jurkat cells and Raji cells in FACS assays;

3) is capable of inhibiting the binding of CD47 to sirpa in an ELISA assay;

4) capable of binding human blood erythrocytes;

5) in the erythrocyte coagulation assay, no coagulation reaction is caused;

6) can cause phagocytosis of tumor cells by macrophages;

7) can inhibit tumor growth and/or tumor cell proliferation.

In certain embodiments, the isolated antigen binding protein comprises an antibody or antigen binding fragment thereof.

In certain embodiments, the antigen binding fragment of the isolated antigen binding protein comprises Fab, Fab', F (ab)₂Fv fragment, F (ab')₂scFv, di-scFv and/or dAb.

In certain embodiments, the antibody in the isolated antigen binding protein is a fully human antibody.

In certain embodiments, the LCDR1 in the isolated antigen binding protein comprises SEQ ID NO: 139.

In certain embodiments, the LCDR1 in the isolated antigen binding protein comprises SEQ ID NO: 113. 114 and 115, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the LCDR2 in the isolated antigen binding protein comprises SEQ ID NO: 140.

In certain embodiments, the LCDR2 in the isolated antigen binding protein comprises SEQ ID NO: 31. 56, 57, 122, 126 and 127.

In certain embodiments, the LCDR3 in the isolated antigen binding protein comprises SEQ ID NO: 141, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the LCDR3 in the isolated antigen binding protein comprises SEQ ID NO: 67-74, 116-121.

In certain embodiments, the HCDR1 in the isolated antigen binding protein comprises SEQ ID NO: 142, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the HCDR1 in the isolated antigen binding protein comprises SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51 and 53.

In certain embodiments, the HCDR2 in the isolated antigen binding protein comprises SEQ ID NO: 143.

In certain embodiments, the HCDR2 in the isolated antigen binding protein comprises SEQ ID NO: 136 and 138.

In certain embodiments, the HCDR3 in the isolated antigen binding protein comprises SEQ ID NO: 144, or a fragment thereof.

In certain embodiments, the HCDR3 in the isolated antigen binding protein comprises SEQ ID NO: 55 and any one of 58-63.

In certain embodiments, the VL comprises the framework regions L-FR1, L-FR2, L-FR3, and L-FR4 in the isolated antigen binding protein.

In certain embodiments, the C-terminus of the L-FR1 in the isolated antigen binding protein is linked directly or indirectly to the N-terminus of the LCDR1, and the L-FR1 comprises the amino acid sequence of SEQ ID NO:149, or a fragment thereof.

In certain embodiments, the L-FR1 in the isolated antigen binding protein comprises SEQ ID NO: 156. 157 and 158.

In certain embodiments, the L-FR2 is located between the LCDR1 and the LCDR2 in the isolated antigen binding protein and the L-FR2 comprises the amino acid sequence of SEQ ID NO: 150.

In certain embodiments, the L-FR2 in the isolated antigen binding protein comprises SEQ ID NO: 191 and 194.

In certain embodiments, the L-FR3 is located between the LCDR2 and the LCDR3 in the isolated antigen binding protein and the L-FR3 comprises the amino acid sequence of SEQ ID NO:151, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the L-FR3 in the isolated antigen binding protein comprises SEQ ID NO: 155. 162 and 166, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the N-terminus of the L-FR4 in the isolated antigen binding protein is linked directly or indirectly to the C-terminus of the LCDR3, and the L-FR4 comprises the amino acid sequence of SEQ ID NO: 152.

In certain embodiments, the L-FR4 in the isolated antigen binding protein comprises SEQ ID NO: 159. 160 and 161.

In certain embodiments, the VL of the isolated antigen binding protein comprises SEQ ID NO: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29.

In certain embodiments, the isolated antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human Ig kappa constant region.

In certain embodiments, the antibody light chain constant region in the isolated antigen binding protein comprises SEQ ID NO:196, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the isolated antigen binding protein comprises an antibody light chain LC, and the LC comprises the amino acid sequence set forth in SEQ ID NO 197.

In certain embodiments, the isolated antigen binding protein comprises an antibody light chain LC, and the LC comprises SEQ ID NO: 2. 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30.

In certain embodiments, the VH of the isolated antigen binding protein comprises the framework regions H-FR1, H-FR2, H-FR3, and H-FR 4.

In certain embodiments, the C-terminus of the H-FR1 in the isolated antigen binding protein is linked directly or indirectly to the N-terminus of the HCDR1, and the H-FR1 comprises the amino acid sequence of SEQ ID NO: 145.

In certain embodiments, the H-FRI in the isolated antigen binding protein comprises SEQ ID NO: 167-170.

In certain embodiments, the H-FR2 in the isolated antigen binding protein is located between the HCDR1 and the HCDR2, and the H-FR2 comprises SEQ ID NO: 146.

In certain embodiments, the H-FR2 in the isolated antigen binding protein comprises SEQ ID NO: 184. 187-190.

In certain embodiments, the H-FR3 in the isolated antigen binding protein is located between the HCDR2 and the HCDR3, and the H-FR3 comprises SEQ ID NO: 147.

In certain embodiments, the H-FR3 in the isolated antigen binding protein comprises SEQ ID NO: 171 and 182.

In certain embodiments, the N-terminus of the H-FR4 in the isolated antigen binding protein is linked directly or indirectly to the C-terminus of the HCDR3, and the H-FR4 comprises the amino acid sequence of SEQ ID NO: 148.

In certain embodiments, the H-FR4 in the isolated antigen binding protein comprises SEQ ID NO: 183. 185 and 186, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the VH in the isolated antigen binding protein comprises SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region comprises a human IgG constant region.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region comprises a human IgG4 constant region.

In certain embodiments, the antibody heavy chain constant region in the isolated antigen binding protein comprises SEQ ID NO:195, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain HC, and the HC comprises SEQ ID NO: 198, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain HC, and the HC comprises SEQ ID NO: 76. 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110 and 112.

In another aspect, the present application provides an isolated one or more nucleic acid molecules encoding an isolated antigen binding protein described herein.

In another aspect, the present application provides a vector comprising a nucleic acid molecule as described herein.

In another aspect, the present application provides a cell comprising a nucleic acid molecule described herein or a vector described herein.

In another aspect, the present application provides a method of making an isolated antigen binding protein described herein, the method comprising culturing a cell described herein under conditions such that the isolated antigen binding protein described herein is expressed.

In another aspect, the present application provides a pharmaceutical composition comprising an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, and/or a cell described herein, and optionally a pharmaceutically acceptable carrier.

In another aspect, the present application provides a use of an isolated antigen binding protein as described herein, a nucleic acid molecule as described herein, a vector as described herein, a cell as described herein, and/or a pharmaceutical composition as described herein, for the manufacture of a medicament for the prevention, alleviation and/or treatment of a tumor.

In certain embodiments, the use of (a), wherein the tumor comprises a solid tumor and/or a hematological tumor.

In another aspect, the present application provides a method of inhibiting the binding of CD47 to sirpa, the method comprising administering an isolated antigen binding protein described herein.

In another aspect, the present application provides a method of preventing, ameliorating and/or treating a tumor comprising administering to a subject in need thereof an isolated antigen binding protein described herein.

Other aspects and advantages of the present application will be readily apparent to those skilled in the art from the following detailed description. Only exemplary embodiments of the present application have been shown and described in the following detailed description. As those skilled in the art will recognize, the disclosure of the present application enables those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention as it is directed to the present application. Accordingly, the descriptions in the drawings and the specification of the present application are illustrative only and not limiting.

Drawings

The specific features of the invention to which this application relates are set forth in the appended claims. The features and advantages of the invention to which this application relates will be better understood by reference to the exemplary embodiments described in detail below and the accompanying drawings. The brief description of the drawings is as follows:

FIGS. 1A-1B show the binding activity (FACS assay) of antigen binding proteins of the present application to Jurkat cells;

FIG. 1C shows the binding activity of the antigen binding proteins PR000806, PR000807 and PR000808 described herein to Raji cells (FACS assay);

FIG. 2 shows the binding activity of the antigen binding proteins PR000796, PR000802, PR000803, PR000804, PR000805, PR000806, PR000807, PR000808 and PR000811 described herein to cynomolgus monkey PBMC cells (FACS assay);

FIG. 3A shows the binding activity (ELISA assay) of the antigen binding proteins PR000796, PR000802, PR000803, PR000804, PR000805, PR000806, PR000807, PR000808 and PR000811 according to the present application to monkey CD47 protein;

FIGS. 3B-3C show the binding activity (ELISA assay) of the antigen binding proteins PR000806, PR000807 and PR000808, respectively, described herein, to human and monkey CD47 protein;

FIGS. 4A-4B show the activity of the antigen binding proteins PR000796, PR000802, PR000803, PR000804, PR000805, PR000806, PR000807, PR000808 and PR000811 described herein to inhibit the binding of human CD47 to SIRPa (ELISA assay);

FIGS. 5A-5B show the activity of the antigen binding proteins PR000796, PR000802, PR000803, PR000804, PR000805, PR000806, PR000807, PR000808 and PR000811 described herein to block the binding of CD47 to SIRPa on the surface of Jurkat cells (FACS assay);

FIGS. 6A-6B show that antigen binding proteins described herein do not cause human erythrocyte agglutination reactions;

FIG. 7 shows the binding capacity of antigen binding proteins of the present application to erythrocytes (FACS assay);

FIGS. 8A-8B show the ability of antigen binding proteins PR000796, PR000802, PR000803, PR000804, PR000805, PR000806, PR000807, PR000808 and PR000811 described herein to cause phagocytosis of Jurkat cells by macrophages;

FIG. 9 shows the ability of antigen binding proteins PR000806, PR000807 and PR000808 described herein to cause phagocytosis of OVCAR3 cells by macrophages;

FIG. 10 shows the ability of antigen binding proteins PR001265, PR001275, PR001276, PR000806, PR000808 and PR001281 described herein to cause phagocytosis of human erythrocytes by macrophages;

FIGS. 11A-11B show the in vivo anti-tumor activity of different concentrations of the antigen binding proteins PR000806 and PR000808 described herein on a NCG-Raji mouse model with changes in tumor volume (A) and mouse body weight (B), respectively;

FIGS. 12A-12B show the in vivo anti-tumor activity of different concentrations of the antigen binding proteins PR001265 and PR000806 as described herein on a B-hSIRPa/hCD47-MC38-hCD47 mouse model with changes in tumor volume (A) and mouse body weight (B), respectively;

FIG. 13 shows the body weight trend of mice before and after treatment with antigen binding proteins PR001265, PR001275, PR001276, PR000806, PR000808 and PR001281 described herein;

FIG. 14 shows the results of routine blood measurements from mouse models 7 days prior to treatment with antigen binding proteins PR001265, PR001275, PR001276, PR000806, PR000808 and PR001281 described herein;

FIG. 15 shows the results of routine blood measurements from day 1 mouse models following treatment with antigen binding proteins PR001265, PR001275, PR001276, PR000806, PR000808 and PR001281 described herein;

FIG. 16 shows the results of routine blood measurements from mouse models at day 7 after treatment with antigen binding proteins PR001265, PR001275, PR001276, PR000806, PR000808 and PR001281 described herein;

figure 17 shows the use of the antigen binding proteins PR001265, PR001275 and PR001276 described herein to bind CD47 with high specificity, but not to the non-CD 47 antigen.

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification.

In the present application, the term "antigen binding protein" generally refers to a protein comprising a portion that binds an antigen, and optionally a scaffold or backbone portion that allows the portion that binds the antigen to adopt a conformation that facilitates binding of the antigen binding protein to the antigen. May typically comprise an antibody light chain variable region (VL), an antibody heavy chain variable region (VH), or both. The VH and VL regions can be further distinguished as hypervariable regions, termed Complementarity Determining Regions (CDRs), interspersed with more conserved regions termed Framework Regions (FRs). Each VH and VL may be composed of three CDRs and four FR regions, which may be arranged from amino-terminus to carboxy-terminus in the following order: FR-1, CDR1, FR-2, CDR2, FR-3, CDR3 and FR-4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. Examples of antigen binding proteins include, but are not limited to, antibodies, antigen binding fragments (Fab, Fab', F (ab))₂Fv fragment, F (ab')₂，scFv，di-scFv and/or dAb), immunoconjugates, multispecific antibodies (e.g., bispecific antibodies), antibody fragments, antibody derivatives, antibody analogs, or fusion proteins, etc., so long as they exhibit the desired antigen-binding activity.

In the present application, the term "variable" generally refers to the fact that certain portions of the sequences of the variable domains of antibodies vary strongly, which results in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable region of the antibody. It is concentrated in three segments in the light and heavy chain variable regions, called Complementarity Determining Regions (CDRs) or hypervariable regions (HVRs). 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, largely in a β -sheet configuration, connected by three CDRs, forming a loop junction, and in some cases forming part of a β -sheet structure. The CDRs in each chain are held in close proximity by the FR regions and together with the CDRs from the other chain form the antigen-binding site of the antibody, and the constant regions are not directly involved in the binding of the antibody to the antigen, but they exhibit different effector functions, e.g., participation in antibody-dependent cytotoxicity of the antibody. In the art, the CDRs of an antibody can be defined by a variety of methods, such as Kabat's rules of definition based on sequence variability (see, Kabat et al, immunological protein sequences, fifth edition, national institute of health, Besserda, Md. (1991)) and Chothia's rules of definition based on the position of the structural loop regions (see, A1-Lazikani et al, Jmol Biol 273: 927-48, 1997). In this application, the rules of Combined definition, including the Kabat definition and the Chothia definition, are also used to determine amino acid residues in variable domain sequences and full-length antibody sequences.

TABLE 1 CDR definition method for the antibodies of the present application (see http:// bio if. org. uk/abs /)

	Kabat	Chothia	Combined
				LCDR1	L24--L34	L24--L34	L24-L34
LCDR2	L50--L56	L50--L56	L50-L56
				LCDR3	L89--L97	L89--L97	L89-L97
HCDR1	H31--H35	H26--H32	H26-H35
				HCDR2	H50--H65	H52--H56	H50-H65
HCDR3	H95--H102	H95--H102	H95-H102

Wherein Laa-Lbb can refer to the amino acid sequence from position aa (Chothia coding rule) to position bb (Chothia coding rule) from the N-terminus of the antibody light chain; Haa-Hbb can refer to the amino acid sequence from position aa (Chothia coding rule) to position bb (Chothia coding rule) from the N-terminus of the antibody heavy chain. For example, L24-L34 may refer to the amino acid sequence from position 24 to position 34, beginning at the N-terminus of the antibody light chain, according to the Chothia coding rules; H26-H32 can refer to the amino acid sequence from position 26 to position 32, beginning at the N-terminus of the antibody heavy chain, according to the Chothia coding rules.

In the present application, the term "isolated antigen binding protein" generally refers to an antigen binding protein that has been identified, isolated and/or recovered from a component of its production environment (e.g., native or recombinant). The contaminating components of the environment that they produce are often substances that interfere with their research, diagnostic or therapeutic uses and may include enzymes, hormones and other proteinaceous or non-proteinaceous solutes. An isolated antigen binding protein or antibody will typically be prepared by at least one purification step.

In the present application, the term "monoclonal antibody" generally refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies in the population are identical except for possible small amounts of natural mutations. Monoclonal antibodies are typically highly specific for a single antigenic site. Moreover, unlike conventional polyclonal antibody preparations (which typically have different antibodies directed against different determinants), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies have the advantage that they can be synthesized by hybridoma culture, uncontaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies for use according to the invention may be prepared in hybridoma cells, or may be prepared by recombinant DNA methods.

In this application, the term "fully human antibody" generally refers to an antibody that is expressed by an animal by transferring all of the human antibody-encoding genes into a genetically engineered antibody gene-deleted animal. All parts of an antibody (including the variable and constant regions of an antibody) are encoded by genes of human origin. The fully human antibody can greatly reduce the immune side reaction of the heterologous antibody to the human body. The method for obtaining fully human antibody in the field can be phage display technology, transgenic mouse technology, ribosome display technology, RNA-polypeptide technology and the like.

In the present application, the term "specific anti-CD 47 antibody" generally refers to an antibody that can compete with the antibodies of the present application for binding to CD47. Antigen binding proteins of the present application may include those antibodies having the same amino acid sequence as the particular anti-CD 47 antibody.

In the present application, the term "specific binding" generally refers to the binding of an antibody to an epitope via its antigen binding domain, and this binding requires some complementarity between the antigen binding domain and the epitope. According to this definition, an antibody is said to "specifically bind" to a random, unrelated epitope when it is more likely to bind to the epitope through its antigen binding domain than it would. An "epitope" refers to a specific group of atoms (e.g., sugar side chain, phosphoryl, sulfonyl) or amino acid on an antigen that binds to an antigen binding protein (e.g., an antibody).

In the present application, the term "CD 47" generally refers to the leukocyte surface antigen CD47, which may be the entire CD47 protein, but also any form of CD47 and variants thereof that retain at least a portion of the activity of CD47. The CD47 may include human CD47 and fragments thereof or monkey CD47 and fragments thereof. For example, human CD47 can comprise the amino acid sequence shown in UniProtKB database under accession No. q08722. The monkey may be a cynomolgus monkey. For example, cynomolgus monkey CD47 may comprise the amino acid sequence of accession No. a0a2k5x412 in the UniProtKB database. In the present application, the terms "human CD47 extracellular domain" and "hcd 47. ecd" are used interchangeably and refer generally to the domain of the CD47 extracellular N-terminal V-type immunoglobulin, which is also the CD47 ligand binding region. For example, the extracellular domain of human CD47 may comprise the 19 th to 141 th bit sequence of the amino acid sequence shown in UniProtKB under accession No. q08722.

In the present application, the term "sirpa" generally refers to a signal-regulating protein α, also known as BIT, MFR, MYD1, PTPNS1, SHPS1 or SIRP, that is an immunoglobulin-like cell surface receptor of CD47, either as an intact sirpa protein or in any form of sirpa and variants thereof that retain at least a partial activity of sirpa. In some cases, the sirpa may be a human sirpa, which may comprise the amino acid sequence shown in UniProtKB database under accession number P78324.

In the present application, the term "Jurkat cells" refers generally to human leukemic T lymphocytes. The Jurkat cells were derived from peripheral blood of a 14 year old boy.

In the present application, the term "Raji cell" generally refers to human Burkitt's lymphoma cells. The Raji cells were derived from Burkitt's lymphoma in the left maxilla of an 11 year old black boy.

In this application, the terms "OVCAR 3 cell" and "OVCAR-3 cell" are used interchangeably and refer generally to human ovarian adenocarcinoma cells. The OVCAR3 cells are derived from malignant ascites in patients with progressive ovarian adenocarcinoma.

In the present application, the terms "coagulation reaction", "agglutination" are used interchangeably and generally refer to the homotypic interaction that occurs when red blood cells agglutinate or clump when incubated with different reagents. The agglutination may be a phenomenon in which agglutination of erythrocytes blurs the edge of a blood disk in the presence of a control antibody. The existing CD47 antibody was reported to cause RBC agglutination at certain concentrations, as the existing antibody Tab1(FortySeven, Hu5F9-G) in example 7 of the present application. Thus, the clotting effects may limit the therapeutic efficacy of the CD47 antibody. The term "does not cause a coagulation response" generally means that there is no agglutination or clumping of red blood cells.

In the present application, the term "Tab 1" generally refers to the FortySeven corporation anti-CD 47 antibody Hu5F 9-G.

In this application, the term "Tab 2" refers generally to the anti-CD 47 antibody CC-90002 from Celgene corporation.

In the present application, the term "Tab 3" generally refers to the anti-CD 47 antibody SRF-231 from Surface Oncology.

In the present application, the terms "KD", "KD" are used interchangeably, and generally refer to the equilibrium dissociation constant, and to the value obtained in a titration measurement, either at equilibrium, or by dividing the dissociation rate constant (koff) by the association rate constant (kon). The binding affinity of an antigen-binding protein (e.g., an antibody) for an antigen can be expressed using an association rate constant (kon), an dissociation rate constant (koff), and an equilibrium dissociation constant (KD). Methods for determining the association and dissociation rate constants are well known in the art. The use of fluorescence-based techniques can provide high sensitivity and the ability to examine the sample at equilibrium in physiological buffer. Other experimental approaches and instruments such as BIAcore (biomolecular interaction analysis) assays may be used.

In the present application, the term "nucleic acid molecule" generally refers to an isolated form of nucleotides, deoxyribonucleotides or ribonucleotides or analogs thereof of any length, isolated from their natural environment or artificially synthesized.

In the present application, the term "vector" generally refers to a nucleic acid molecule capable of self-replication in a suitable host, which transfers the inserted nucleic acid molecule into and/or between host cells. The vector may include a vector mainly for inserting a DNA or RNA into a cell, a vector mainly for replicating a DNA or RNA, and a vector mainly for expression of transcription and/or translation of a DNA or RNA. The vector also includes vectors having a plurality of the above-described functions. The vector may be a polynucleotide capable of being transcribed and translated into a polypeptide when introduced into a suitable host cell. Typically, the vector will produce the desired expression product by culturing a suitable host cell containing the vector.

In the present application, the term "cell" generally refers to an individual cell, cell line or cell culture that may or may already contain a plasmid or vector comprising a nucleic acid molecule described herein, or that is capable of expressing an antibody or antigen-binding fragment thereof described herein. The cell may comprise progeny of a single host cell. Due to natural, accidental, or deliberate mutation, the progeny cells may not necessarily be identical in morphology or in genome to the original parent cell, but may be capable of expressing an antibody or antigen-binding fragment thereof as described herein. The cells can be obtained by in vitro transfection of cells using the vectors described herein. The cell may be a prokaryotic cell (e.g., E.coli) or a eukaryotic cell (e.g., a yeast cell, such as a COS cell, a Chinese Hamster Ovary (CHO) cell, a HeLa cell, a HEK293 cell, a COS-1 cell, an NS0 cell, or a myeloma cell). In some embodiments, the cell is a mammalian cell. For example, the mammalian cell may be a CHO-K1 cell. In the present application, the term "recombinant cell" generally refers to a cell into which a recombinant expression vector has been introduced. The recombinant host cell includes not only a specific cell but also a progeny of such a cell.

In the present application, the term "pharmaceutical composition" generally refers to a formulation that is present in a form that allows the biological activity of the active ingredient to be effective, and that does not contain additional ingredients that have unacceptable toxicity to the subject to which the composition will be administered. The composition is sterile. "sterile" compositions are sterilized or free of all living microorganisms and their spores.

In the present application, the term "tumor" generally refers to a neoplasm formed by local tissue cell proliferation in a body (e.g., cells or components thereof) of a mammal under the action of various tumorigenic factors. In the present application, tumors may include lymphomas, leukemias, and ovarian cancers. For example, the tumor may include invasive non-hodgkin's lymphoma, burkitt's lymphoma, Acute Lymphocytic Leukemia (ALL), Acute Myelogenous Leukemia (AML), Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML), ovarian cancer, and the like.

In this application, the term "between … …" generally means that the C-terminus of an amino acid fragment is directly or indirectly linked to the N-terminus of a first amino acid fragment and that the N-terminus is directly or indirectly linked to the C-terminus of a second amino acid fragment. In the light chain, for example, the N-terminus of the L-FR2 is linked directly or indirectly to the C-terminus of the LCDR1, and the C-terminus of the L-FR2 is linked directly or indirectly to the N-terminus of the LCDR 2. For another example, the N-terminus of the L-FR3 is directly or indirectly linked to the C-terminus of the LCDR2, and the C-terminus of the L-FR3 is directly or indirectly linked to the N-terminus of the LCDR 3. In the heavy chain, for example, the N-terminus of the H-FR2 is linked directly or indirectly to the C-terminus of the HCDR1, and the C-terminus of the H-FR2 is linked directly or indirectly to the N-terminus of the HCDR 2. For another example, the N-terminus of the H-FR3 is directly or indirectly linked to the C-terminus of the HCDR2, and the C-terminus of the H-FR3 is directly or indirectly linked to the N-terminus of the HCDR 3. In the present application, the "first amino acid fragment" and the "second amino acid fragment" may be any one of the same or different amino acid fragments.

In the present application, the term "comprising" is generally intended to include the explicitly specified features, but not to exclude other elements.

In the present application, the term "about" generally means varying from 0.5% to 10% above or below the stated value, for example, varying from 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below the stated value.

Antibodies, antigen-binding fragments or variants thereof

In one aspect, the present application provides an antigen binding protein comprising at least one CDR in a light chain variable region VL, which VL may comprise the amino acid sequence of SEQ ID NO: 153.

In the present application, the antigen binding protein may comprise LCDR1, and the LCDR1 may comprise SEQ ID NO: 139 with the amino acid sequence:

RASQSX₁SSX₂LX₃(SEQ ID NO: 139); wherein, X₁I or V; x₂W, Y or H; x₃A or N. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 113, said LCDR1 may be included in at least one protein selected from the group consisting of LCDR1Substitutions at the following amino acid positions: at X₁、X₂And/or X₃Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 113, said LCDR1 may be comprised at least at X in comparison to LCDR1 of the antigen binding protein as set forth in₁、X₂Or X₃Wherein the amino acid at X1 can be substituted with V; x₂The amino acid at (A) may be substituted with Y or H; x₃The amino acid at (a) may be substituted with N.

For example, the LCDR1 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 113-115.

In the present application, the antigen binding protein may comprise LCDR2, and the LCDR2 may comprise SEQ ID NO: 140, and the amino acid sequence shown as seq id no:

X₁X₂SX₃X₄X₅X₆(SEQ ID NO: 140); wherein, X₁K, R, V or G; x₂A or S; x₃(ii) S, T, R or I; x₄L or R; x₅E, D, Q or a; x₆S or T. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 56, the LCDR2 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X1, X₂、X₃、X₄、X₅And/or X₆Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 56, said LCDR2 may be comprised at least at X as compared to LCDR2 of the antigen binding protein set forth in₁、X₂、X₃、X₄、X₅Or X₆Amino acid substitution of (a), wherein X₁The amino acid at (b) may be substituted with R, V or G; x₂The amino acid at (a) may be substituted with S; x₃The amino acid at (b) may be substituted with R, I or T; x₄The amino acid at (a) may be substituted with R; x₅The amino acid at (a) may be substituted with D, Q or a; x₆The amino acid at (a) may be substituted with T.

For example, the LCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 31. 56, 57, 122, 126 and 127.

In the present application, the antigen binding protein may comprise LCDR3, and the LCDR3 may comprise SEQ ID NO: 141, and the amino acid sequence shown in seq id no:

X₁X₂X₃X₄X₅X₆X₇X₈X₉X₁₀t (SEQ ID NO: 141); wherein, X₁Q or R; x₂Q, L or H; x₃-Y or-; x₄N, I, Q or-; x₅(ii) S, Y, T or N; x₆Y, W, N or Q; x₇-S, T or-; x₈-T or-; x₉P or-; x₁₀Y, R or L. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 116, the LCDR3 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉And/or X₁₀Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 116, said LCDR3 may be comprised at least at X in comparison to LCDR3 of the antigen binding protein as set forth in₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉And/or X₁₀Amino acid substitution of (a), wherein X₁The amino acid at (a) may be substituted with Q; x₂The amino acid at (A) may be substituted with L or H; x₃The amino acid at (a) may be substituted with no amino acid; x₄The amino acid at (b) may be substituted with I, Q or no amino acid; x₅The amino acid at (b) may be substituted with Y, T or N; x₆The amino acid at (b) may be substituted with Q, N or W; x₇The amino acid at (a) may be substituted with T or no amino acid; x₈No amino acid at (a) may be substituted with T; x₉The amino acid at (a) may be substituted with no amino acid; x₁₀The amino acid at (A) may be substituted with R or L.

In this application, "-" indicates that this position may be free of amino acid substitutions as compared to the LCDR3 of the control antigen binding protein. For example, a peptide similar to SEQ ID NO: 116 LCDR3 (X) of the antigen binding protein₉P) compared to SEQ ID NO: LCDR3 of antigen binding protein shown at 74 at X₉There are no amino acid substitutions.

For example, the LCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 67-74, 116 and 121.

The antigen binding proteins described herein may comprise at least one CDR in an antibody heavy chain variable region VH comprising the amino acid sequence of SEQ ID NO: 154.

In the present application, the antigen binding protein may comprise HCDR1, and the HCDR1 may comprise SEQ ID NO: 142:

GX₁SX₂X₃X₄y (SEQ ID NO: 142); wherein, X₁G or D; x₂I, L or V; x₃(ii) S, N, D or Q; x₄N, S or T. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 40, said HCDR1 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃And/or X₄Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 40, said HCDR1 may be comprised at least at X as compared to HCDR1 of the antigen binding protein set forth in₁、X₂、X₃And/or X₄Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with D; x₂The amino acid at (A) may be substituted with L or V; x₃The amino acid at (b) may be substituted with S, D or Q; x₄The amino acid at (A) may be substituted with S or T.

For example, the HCDR1 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51 and 53.

In the present application, the antigen binding protein may comprise HCDR2, and the HCDR2 may comprise SEQ ID NO: 143 by seq id no:

YYSGX₁(SEQ ID NO: 143); wherein, X₁T, N or S. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 136, the HCDR2 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 136, said HCDR2 may be comprised at least at X compared to HCDR2 of an antibody as set forth in₁Amino acid substitution of (a), wherein, X₂The amino acid at (A) may be substituted with S or T.

For example, the HCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 136-138.

In the present application, the antigen binding protein may comprise HCDR3, and the HCDR3 may comprise SEQ ID NO: 144, and the amino acid sequence shown in SEQ ID NO:

X₁X₂X₃X₄X₅X₆y (SEQ ID NO: 144); wherein, X₁K or G; x₂R or K; x₃G, a, T, W or L; x₄V, M, G, I or a; x₅L, Y, H, F or-; x₆D or S. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 60, said HCDR3 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅And/or X₆Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 60, the HCDR3 may be comprised at least at X₁、X₂、X₃、X₄、X₅And/or X₆OfAmino acid substitution, wherein, X₁The amino acid at (a) may be substituted with G; x₂The amino acid at (a) may be substituted with K; x₃The amino acid at (b) may be substituted with A, T, L or W; x₄The amino acid at (b) may be substituted with M, G, A or I; x₅The amino acid at (b) may be substituted with Y, H, F or no amino acid; x₆The amino acid at (a) may be substituted with S.

For example, the HCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 55. 58-63.

In the present application, the isolated antigen binding protein may comprise LCDR 1-3. The LCDR1 can comprise SEQ ID NO: 139; the LCDR2 can comprise SEQ ID NO: 140; the LCDR3 can comprise SEQ ID NO: 141, or a pharmaceutically acceptable salt thereof.

In certain instances, the LCDR1 of an antigen binding protein described herein can comprise an amino acid sequence set forth in any one of: SEQ ID NO: 113. 114 and 115; the LCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 31. 56, 57, 122, 126 and 127; the LCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 67-74, 116 and 121.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 113; LCDR2 may comprise SEQ ID NO: 56; LCDR3 may comprise SEQ ID NO: 116.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 113; LCDR2 may comprise SEQ ID NO: 122; LCDR3 may comprise SEQ ID NO: 73, or a pharmaceutically acceptable salt thereof.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 114; LCDR2 may comprise SEQ ID NO: 126, or a pharmaceutically acceptable salt thereof; LCDR3 may comprise SEQ ID NO: 72, or a pharmaceutically acceptable salt thereof.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 115; LCDR2 may comprise SEQ ID NO: 31; LCDR3 may comprise SEQ ID NO: 70.

In the present application, the isolated antigen binding protein may comprise HCDR 1-3. The HCDR1 can comprise SEQ ID NO: 142; the HCDR2 can comprise SEQ ID NO: 143; and the HCDR3 may comprise SEQ ID NO: 144, or a fragment thereof.

In certain instances, the HCDR1 of an antigen binding protein described herein can comprise an amino acid sequence set forth in any one of: SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51, and 53; the HCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 136-138; and the HCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 58-63 and 55.

For example, HCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 34; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 58.

For example, HCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 42; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 59, or a pharmaceutically acceptable salt thereof.

For example, HCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 38; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 55.

For example, HCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 36; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 58.

For example, HCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 42; HCDR2 can comprise SEQ ID NO: 136; and HCDR3 can comprise SEQ ID NO: 60, or a pharmaceutically acceptable salt thereof.

In the present application, the isolated antigen binding protein may comprise LCDR1-3 and HCDR 1-3. The LCDR1 can comprise SEQ ID NO: 139; the LCDR2 can comprise SEQ ID NO: 140; the LCDR3 can comprise SEQ ID NO: 141; the HCDR1 can comprise SEQ ID NO: 142; the HCDR2 can comprise SEQ ID NO: 143; and the HCDR3 may comprise SEQ ID NO: 144, or a fragment thereof.

In certain instances, the LCDR1 of an antigen binding protein described herein can comprise an amino acid sequence set forth in any one of: SEQ ID NO: 113. 114 and 115; the LCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 31. 56, 57, 122, 126 and 127; the LCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 67-74, 116-121; the HCDR1 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51, and 53; the HCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 136-138; and the HCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 58-63 and 55.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 113; LCDR2 may comprise SEQ ID NO: 122; LCDR3 may comprise SEQ ID NO: 73; its HCDR1 may comprise SEQ ID NO: 42; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 59, or a pharmaceutically acceptable salt thereof.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 114; LCDR2 may comprise SEQ ID NO: 126, or a pharmaceutically acceptable salt thereof; LCDR3 may comprise SEQ ID NO: 72; its HCDR1 may comprise SEQ ID NO: 38; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 55.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 115; LCDR2 may comprise SEQ ID NO: 31; LCDR3 may comprise SEQ ID NO: 70; its HCDR1 may comprise SEQ ID NO: 36; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 58.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 113; LCDR2 may comprise SEQ ID NO: 56; LCDR3 may comprise SEQ ID NO: 116; its HCDR1 may comprise SEQ ID NO: 45, or a pharmaceutically acceptable salt thereof; HCDR2 can comprise SEQ ID NO: 138; and HCDR3 can comprise SEQ ID NO: 60, or a pharmaceutically acceptable salt thereof.

For example, LCDR1 in the antigen binding proteins described herein can comprise SEQ ID NO: 113; LCDR2 may comprise SEQ ID NO: 56; LCDR3 may comprise SEQ ID NO: 74; its HCDR1 may comprise SEQ ID NO: 53; HCDR2 can comprise SEQ ID NO: 137; and HCDR3 can comprise SEQ ID NO: 63.

The antigen binding proteins described herein may also comprise the framework regions L-FR1, L-FR2, L-FR3 and L-FR 4.

In the present application, the antigen binding protein may comprise L-FR1, the C-terminus of L-FR1 is linked directly or indirectly to the N-terminus of LCDR1, and the L-FR1 comprises the amino acid sequence of SEQ ID NO:149 of seq id no:

X₁IX₂MTQSPX₃X₄LSX₅SX₆GX₇RX₈TX₉X₁₀c (SEQ ID NO: 149); wherein, X₁D or E; x₂Q or V; x₃(ii) S or a; x₄T or S; x₅A or V; x₆V or P; x₇D or E; x₈V or a; x₉I or L; x₁₀T or S. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 157, said L-FR1 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉And/or X₁₀Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 157 in comparison with L-FR1 of the antigen-binding protein shown in 157, said L-FR1 may be contained at least in X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉And/or X₁₀Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with E; x₂The amino acid at (a) may be substituted with V; x₃The amino acid at (A) may be substituted with A; x₄The amino acid at (a) may be substituted with S; x₅The amino acid at (a) may be substituted with V; x₆The amino acid at (a) may be substituted with P; x₇The amino acid at (a) may be substituted with E; x₈The amino acid at (A) may be substituted with A; x₉The amino acid at (a) may be substituted with L; x₁₀The amino acid at (a) may be substituted with S.

For example, the L-FR1 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 156-158.

In the present application, the antigen binding protein may comprise L-FR2, the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises the amino acid sequence of SEQ ID NO: 150:

WYQQKPGX₁APX₂LLIY (SEQ ID NO: 150); wherein, X₁＝K，NOr Q; x₂K or R. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 191 the antigen binding protein of L-FR2 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁And/or X₂Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 191 the antigen binding protein may comprise at least L-FR2 as compared to L-FR2 of the antigen binding protein as represented by 191 at least₁And/or X₂Amino acid substitution of (a), wherein, X₁The amino acid at (A) may be substituted with N or Q; x₂The amino acid at (a) may be substituted with R.

For example, the L-FR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 191-194.

In the present application, the antigen binding protein may comprise L-FR3, the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises the amino acid sequence of SEQ ID NO:151, and (b) the amino acid sequence shown in (a):

X₁X₂PX₃RFSGSGSGTX₄FX₅LTX₆SSLQX₇X₈DFAX₉YYC (SEQ ID NO: 151); wherein, X₁＝G，D；X₂V or I; x₃(ii) S or V; x₄E or D; x₅T or S; x₆I or V; x₇P or S; x₈D or E; x₉T or I. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 166 can comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈And/or X₉Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 166 can be contained at least in X as compared with L-FR3 of the antibody represented by 166₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈And/or X₉Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with D; x₂The amino acid at (A) may be substituted with I; x₃The amino acid at (a) may be substituted with V; x₄The amino acid at (a) may be substituted with D; x₅The amino acid at (a) may be substituted with S; x₆The amino acid at (a) may be substituted with V; x₇The amino acid at (a) may be substituted with S; x₈The amino acid at (a) may be substituted with E; x₉The amino acid at (A) may be substituted with I.

For example, the L-FR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 155. 162, and 166.

In the present application, the antigen binding protein may comprise L-FR4, the N-terminus of L-FR4 is linked to the C-terminus of LCDR3, and the L-FR4 comprises the amino acid sequence of SEQ ID NO: 152:

FGX₁GTKX₂EIK (SEQ ID NO: 152); wherein, X₁Q or G; x₂L or V. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 160, said L-FR4 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁And/or X₂Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 160, the L-FR4 may be at least one of the L-FR4 that is at least one of the L-FR4 that is at least one of the L-FR4₁And/or X₂Wherein the amino acid at X1 can be substituted with G; the amino acid at X2 may be substituted with V.

For example, the L-FR4 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 159-161.

In the present application, the antigen binding protein may comprise a light chain variable region VL, and the VL may comprise SEQ ID NO: 153:

X₁IX₂MTQSPX₃X₄LSX₅SX₆GX₇RX₈TX₉X₁₀CRASQSX₁₁SSX₁₂LX₁₃WYQQKPGX₁₄APX₁₅LLIYX₁₆X₁₇SX₁₈X₁₉X₂₀X₂₁X₂₂X₂₃PX₂₄RFSGSGSGTX₂₅FX₂₆LTX₂₇SSLQX₂₈X₂₉DFAX₃₀YYCX₃₁X₃₂X₃₃X₃₄X₃₅X₃₆X₃₇X₃₈X₃ ₉X₄₀TFGX₄₁GTKX₄₂EIK (SEQ ID NO: 153); wherein, X₁＝D，E；X₂＝Q，V；X₃(ii) S or a; x₄T or S; x₅A or V; x₆V or P; x₇D or E; x₈V or a; x₉I or L; x₁₀T or S; x₁₁I or V; x₁₂W or Y or H; x₁₃A or N; x₁₄K, N or Q; x₁₅K or R; x₁₆K, R, V or G; x₁₇A or S; x₁₈(ii) S, T, R or I; x₁₉L or R; x₂₀E, D, Q or a; x₂₁(ii) S or T; x₂₂G or D; x₂₃V or I; x₂₄(ii) S or V; x₂₅E or D; x₂₆T or S; x₂₇I or V; x₂₈P or S; x₂₉D or E; x₃₀T or I; x₃₁Q or R; x₃₂Q, L or H; x₃₃-Y or-; x₃₄N, I, Q or-; x₃₅(ii) S, T or N; x₃₆Y, W, N or Q; x₃₇-S, T or-; x₃₈-T or-; x₃₉P or-; x₄₀Y, R or L; x₄₁Q or G; x₄₂L or V. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO:15, the VL may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄、X₁₅、X₁₆、X₁₇、X₁₈、X₁₉、X₂₀、X₂₁、X₂₂、X₂₃、X₂₄、X₂₅、X₂₆、X₂₇、X₂₈、X₂₉、X₃₀、X₃₁、X₃₂、X₃₃、X₃₄、X₃₅、X₃₆、X₃₇、X₃₈、X₃₉、X₄₀、X₄₁And/or X₄₂Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO:15, said VL may be comprised at least in X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄、X₁₅、X₁₆、X₁₇、X₁₈、X₁₉、X₂₀、X₂₁、X₂₂、X₂₃、X₂₄、X₂₅、X₂₆、X₂₇、X₂₈、X₂₉、X₃₀、X₃₁、X₃₂、X₃₃、X₃₄、X₃₅、X₃₆、X₃₇、X₃₈、X₃₉、X₄₀、X₄₁And/or X₄₂Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with E; x₂The amino acid at (a) may be substituted with V; x₃The amino acid at (A) may be substituted with A; x₄The amino acid at (a) may be substituted with S; x₅The amino acid at (a) may be substituted with V; x₆The amino acid at (a) may be substituted with P; x₇The amino acid at (a) may be substituted with E; x₈The amino acid at (A) may be substituted with A; x₉The amino acid at (a) may be substituted with L; x₁₀The amino acid at (a) may be substituted with S; x₁₁The amino acid at (a) may be substituted with V; x₁₂The amino acid at (A) may be substituted with H or Y; x₁₃The amino acid at (a) may be substituted with N; x₁₄The amino acid at (A) may be substituted with Q or N; x₁₅The amino acid at (a) may be substituted with R; x₁₆The amino acid at (b) may be substituted with G, R or V; x₁₇The amino acid at (a) may be substituted with S; x₁₈The amino acid at (b) may be substituted with I, R or T; x₁₉The amino acid at (a) may be substituted with R; x₂₀The amino acid at (b) may be substituted with A, Q or D; x₂₁The amino acid at (a) may be substituted with T; x₂₂The amino acid at (a) may be substituted with D; x₂₃The amino acid at (A) may be substituted with I; x₂₄The amino acid at (a) may be substituted with V; x₂₅The amino acid at (a) may be substituted with D; x₂₆The amino acid at (a) may be substituted with S; x₂₇The amino acid at (a) may be substituted with V; x₂₈The amino acid at (a) may be substituted with S; x₂₉The amino acid at (a) may be substituted with E; x₃₀The amino acid at (A) may be substituted with I; x₃₁The amino acid at (a) may be substituted with Q; x₃₂The amino acid at (A) may be substituted with L or H; x₃₃The amino acid at (a) may be substituted with no amino acid; x₃₄The amino acid at (A) may be substituted with I or without an amino acid; x₃₅The amino acid at (b) may be substituted with Y, T or N; x₃₆The amino acid at (a) may be substituted with W or N; x₃₇The amino acid at (a) may be substituted with no amino acid; x₃₈The amino acid at (a) may be substituted with T; x₃₉The amino acid at (a) may be substituted with no amino acid; x₄₀The amino acid at (A) may be substituted with L or R; x₄₁The amino acid at (a) may be substituted with G; x₄₂The amino acid at (a) may be substituted with V.

For example, the VL region may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29.

The antigen binding proteins described herein may also comprise the framework regions H-FR1, H-FR2, H-FR3, and H-FR 4.

In the present application, the antigen binding protein may comprise H-FR1, the C-terminus of H-FR1 is linked directly or indirectly to the N-terminus of HCDR1, and the H-FR1 comprises SEQ ID NO:145 by amino acid sequence:

QVQLQESGPGLX₁X₂PSETLSLTCX₃VS (SEQ ID NO: 145); wherein, X₁V or M; x₂K or Q; x₃T or S. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 169 compared to H-FR1 of the antigen binding protein, said H-FR1 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂And/or X₃Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 169 the H-FR1 may be contained at least in X as compared with H-FR1 shown in₁、X₂And/or X₃Wherein the amino acid at X1 can be substituted with M; x₂The amino acid at (a) may be substituted with Q; x₃The amino acid at (a) may be substituted with S.

For example, the H-FR1 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 167-170.

In the present application, the antigen binding protein may comprise H-FR2, the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises SEQ ID NO: 146:

YWX₁WX₂RQX₃PGX₄GLEWIGX₅i (SEQ ID NO: 146); wherein, X1 ═ S or T; x₂I or F; x₃P or a; x₄K or R; x₅Y or N. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 188 may comprise at least a substitution at an amino acid position selected from the group consisting of: is contained in X₁、X₂、X₃、X₄And/or X₅Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 188 by comparison with H-FR2 shown in the specification, the H-FR2 can be at least contained in X₁、X₂、X₃、X₄And/or X₅Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with T; x₂The amino acid at (a) may be substituted with F; x₃The amino acid at (A) may be substituted with A; x₃The amino acid at (a) may be substituted with R; x₃The amino acid at (a) may be substituted with N.

For example, the H-FR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 184. 187 and 190.

In the present application, the antigen binding protein may comprise H-FR3, the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises SEQ ID NO: 147:

RVX₁X₂SX₃DX₄SKNQFSLX₅LX₆SVX₇X₈X₉DTAX₁₀YYCX₁₁r (SEQ ID NO: 147); wherein, X₁T or S; x₂I or M; x₃V or I; x₄T or I; x₅K or N; x₆(ii) S, T or N; x₇T or I; x₈A or S; x₉A or G; x₁₀V or M; x₁₁A or G. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 178, said H-FR3 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄And/or X₁₅Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 178, said H-FR3 may be at least comprised in X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄And/or X₁₃Wherein the amino acid at X1 can be substituted with D; x₂The amino acid at (a) may be substituted with Q; x₃The amino acid at (a) may be substituted with P; x₄The amino acid at (a) may be substituted with T; x₅The amino acid at (a) may be substituted with S; x₆The amino acid at (a) may be substituted with M; x₇The amino acid at (A) may be substituted with I; x₈The amino acid at (A) may be substituted with I; x₉The amino acid at (a) may be substituted with K; x₁₀The amino acid at (A) may be substituted with S or N; x₁₁The amino acid at (A) may be substituted with I; x₁₂The amino acid at (a) may be substituted with S; x₁₃The amino acid at (a) may be substituted with G; x₁₄The amino acid at (a) may be substituted with M; x₁₅The amino acid at (A) may be substituted with G.

For example, the H-FR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 171-182.

In the present application, the antigen binding protein may comprise H-FR4, the N-terminus of H-FR4 is linked to the C-terminus of HCDR3, and the H-FR4 comprises SEQ ID NO:148, and the amino acid sequence shown in SEQ ID NO:

X₁GQGX₂LVTVSS (SEQ ID NO: 148); wherein, X₁W or S; x₂T or I. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO:186 may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁And/or X₂Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO:186 may be contained at least in X of H-FR4 as compared with H-FR4 shown in₁And/or X₂Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with S; x₂The amino acid at (A) may be substituted with I.

For example, the H-FR4 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 183. 185, and 186.

In the present application, the antigen binding protein may comprise a heavy chain variable region VH, and the VH comprises SEQ ID NO: 154:

QVQLQESGPGLX₁X₂PSETLSLTCX₃VSGX₄SX₅X₆X₇YYWX₈WX₉RQX₁₀PGX₁₁GLEWIGX₁₂IYYSGX₁₃TX₁₄YX₁₅X₁₆SLX₁₇SRVX₁₈X₁₉SX₂₀DX₂₁SKNQFSLX₂₂LX₂₃SVX₂₄X₂₅X₂₆DTAX₂₇YYCX₂₈RX₂₉X₃₀X₃₁X₃₂X₃₃X₃₄YX₃₅GQGX₃₆LVTVSS (SEQ ID NO: 154); wherein, X₁V or M; x₂K or Q; x₃T or S; x₄G or D; x₅I, L or V; x₆(ii) S, N or D; x₇N, S or T; x₈(ii) S or T; x₉I or F; x₁₀P or a; x₁₁K or R; x₁₂Y or N; x₁₃T, S or N; x₁₄N or D; x₁₅N or Q; x₁₆P or S; x₁₇K or T; x₁₈T or S; x₁₉I or M; x₂₀V or I; x₂₁T or I; x₂₂K or N; x₂₃(ii) S, T or N; x₂₄T or I; x₂₅A or S; x₂₆A or G; x₂₇V or M; x₂₈A or G; x₂₉K or G; x₃₀R or K; x₃₁G, a, T, W or L; x₃₂V, M, G, I or a; x₃₃L, Y, H, F or-; x₃₄D or S; x₃₅W or S; x₃₆T or I. For example, the sequence may be a sequence determined according to the Chothia definition rules.

In the present application, the amino acid sequence shown in SEQ ID NO: 85, said VH may comprise at least a substitution at an amino acid position selected from the group consisting of: at X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄、X₁₅、X₁₆、X₁₇、X₁₈、X₁₉、X₂₀、X₂₁、X₂₂、X₂₃、X₂₄、X₂₅、X₂₆、X₂₇、X₂₈、X₂₉、X₃₀、X₃₁、X₃₂、X₃₃、X₃₄、X₃₅And/or X₃₆Amino acid substitution(s) of (c).

In the present application, the amino acid sequence shown in SEQ ID NO: 85 may be comprised at least in X₁、X₂、X₃、X₄、X₅、X₆、X₇、X₈、X₉、X₁₀、X₁₁、X₁₂、X₁₃、X₁₄、X₁₅、X₁₆、X₁₇、X₁₈、X₁₉、X₂₀、X₂₁、X₂₂、X₂₃、X₂₄、X₂₅、X₂₆、X₂₇、X₂₈、X₂₉、X₃₀、X₃₁、X₃₂、X₃₃、X₃₄、X₃₅And/or X₃₆Amino acid substitution of (a), wherein, X₁The amino acid at (a) may be substituted with V; x₂The amino acid at (a) may be substituted with Q; x₃The amino acid at (a) may be substituted with S; x₄The amino acid at (a) may be substituted with D; x₅The amino acid at (A) may be substituted with L or V; x₆The amino acid at (b) may be substituted with S, Q or D; x₇The amino acid at (a) may be substituted with T; x₈The amino acid at (a) may be substituted with T; x₉The amino acid at (a) may be substituted with F; x₁₀The amino acid at (A) may be substituted with A; x₁₁The amino acid at (a) may be substituted with R; x₁₂The amino acid at (a) may be substituted with N; x₁₃The amino acid at (A) may be substituted with T or S; x₁₄The amino acid at (a) may be substituted with D; x₁₅The amino acid at (a) may be substituted with Q; x₁₆The amino acid at (a) may be substituted with P; x₁₇The amino acid at (a) may be substituted with T; x₁₈The amino acid at (a) may be substituted with S; x₁₉The amino acid at (a) may be substituted with M; x₂₀The amino acid at (A) may be substituted with I; x₂₁The amino acid at (A) may be substituted with I; x₂₂The amino acid at (a) may be substituted with K; x₂₃The amino acid at (A) may be substituted with N or T; x₂₄The amino acid at (A) may be substituted with I; x₂₅The amino acid at (a) may be substituted with S; x₂₆The amino acid at (a) may be substituted with G; x₂₇The amino acid at (a) may be substituted with M; x₂₈The amino acid at (a) may be substituted with G; x₂₉The amino acid at (a) may be substituted with G; x₃₀The amino acid at (a) may be substituted with K; x₃₁The amino acid at (b) may be substituted with T, A, W or L; x₃₂The amino acid at (a) may be substituted with I, M, G or a; x₃₃The amino acid at (b) may be substituted with H, F, Y or no amino acid; x₃₄The amino acid at (a) may be substituted with S; x₃₅The amino acid at (a) may be substituted with S; x₃₆The amino acid at (A) may be substituted with I.

For example, the VH may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111.

In the present application, the antigen binding protein may comprise an antibody light chain variable region VL and a heavy chain variable region VH. The VL can comprise SEQ ID NO: 153; the VH may comprise SEQ ID NO: 154.

The VL of an antigen binding protein described herein may comprise an amino acid sequence set forth in any one of SEQ ID NOs: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29; the VH may comprise the amino acid sequence SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:29 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 111.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:29 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 107.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:29 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 109, or a pharmaceutically acceptable salt thereof.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:1, and said VH may comprise the amino acid sequence shown in SEQ ID NO: 79, or a pharmaceutically acceptable salt thereof.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 81. 83, 85, 89, 91, 93, 95 and 97.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 81.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 83 is shown in the figure.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 85, or a pharmaceutically acceptable salt thereof.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 89.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 91.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 93, or a pharmaceutically acceptable salt thereof.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 95, or a pharmaceutically acceptable salt thereof.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO:15 and the VH may comprise the amino acid sequence shown in SEQ ID NO: 97 to any one of the amino acid sequences set forth in seq id No. 97.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO: 23, and said VH may comprise the amino acid sequence shown in SEQ ID NO: 99.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO: 21, and said VH may comprise the amino acid sequence shown in SEQ ID NO: 99.

For example, the isolated antigen binding protein may comprise a VL and a VH, wherein the VL comprises SEQ ID NO: 25, and said VH may comprise the amino acid sequence shown in SEQ ID NO: 99.

In the present application, the antigen binding protein comprises a light chain constant region CL, and the antibody light chain constant region may comprise a human Ig kappa constant region.

For example, the CL region may comprise the amino acid sequence shown below: SEQ ID NO: 196.

in the present application, the antigen binding protein comprises a light chain LC, and the LC may comprise the amino acid sequence of SEQ ID NO: 197, or a pharmaceutically acceptable salt thereof.

For example, the LC may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 2. 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30.

In the present application, the antigen binding protein comprises a heavy chain constant region CH, and the antibody heavy chain constant region may comprise a human IgG constant region. In certain instances, the antibody heavy chain constant region described herein can include a human IgG4 constant region.

For example, the CH region may comprise the amino acid sequence shown below: SEQ ID NO: 195.

in the present application, the antigen binding protein comprises a light chain HC, and the antibody HC may comprise SEQ ID NO: 198, or a pharmaceutically acceptable salt thereof.

For example, the HC may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 76. 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110 and 112.

The antigen binding proteins described herein comprise an antibody light chain constant region CL and a heavy chain constant region CH. The CL can comprise SEQ ID NO: 196; and the CH may comprise SEQ ID NO:195, or a pharmaceutically acceptable salt thereof.

The antigen binding proteins described herein comprise an antibody light chain LC and a heavy chain HC. The LC may comprise SEQ ID NO: 197, and the HC may comprise the amino acid sequence shown in SEQ ID NO: 198, or a pharmaceutically acceptable salt thereof.

For example, the LC may comprise the amino acid sequence SEQ ID NO: 2. 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, and 30, the HC may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 76. 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110 and 112.

The antigen binding proteins described herein may comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 139, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 140, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 141 and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:149, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:150, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:151, and L-FR4 may comprise the amino acid sequence set forth in SEQ ID NO: 152. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 153, and said CL may comprise the amino acid sequence of SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 197, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 142, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 143, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 144, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:145, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:146, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO 147, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 148. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 154, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 198, or a pharmaceutically acceptable salt thereof.

In some cases, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise the amino acid sequence set forth in SEQ ID NO: 113, 115, LCDR2 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 31. 56, 57, 122, 126, and 127, LCDR3 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 67-74, 116-121, said L-FR1 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 156-158, L-FR2 may comprise the amino acid sequence shown in any one of SEQ ID NOs: 191-194, L-FR3 may comprise the amino acid sequence shown in any one of SEQ ID NO: 155. 162, 166, and L-FR4 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 159-161. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29, and the CL may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 2. 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51 and 53, the HCDR2 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 136 and 138, HCDR3 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 55 and 58-63, said H-FR1 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 167-170, H-FR2 may comprise the amino acid sequence shown in any one of SEQ ID NOs: 184. 187-190, H-FR3 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 171 and 182, and H-FR4 may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 183. 185 and 186, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111, the CH may comprise the amino acid sequence set forth in any one of SEQ ID NOs: 195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 76. 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110 and 112.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 74, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 162, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:5, the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 6. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 53, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 63, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 182, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 183. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 105, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 106. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001281.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 122, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 73, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 193, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 27, and said CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 28, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 59, said H-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:186, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 172, and H-FR4 may comprise the amino acid sequence set forth in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 87, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 88, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000806.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 114, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 126, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 72, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 156, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 163, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:1, the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO:2, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 38, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 55, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 184, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 180, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 79, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 80, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000808.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 114, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 127, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 71, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 156, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO: 192, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 164, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 161. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:3, the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 4. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 61, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 167, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 189, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 171, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 75, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 76. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000807.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO:115, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:31, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:70, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:158, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:194, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:155, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:159, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:29, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 30. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO:32, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:58, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:170, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:190, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:176, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 107, and the CH may comprise the amino acid sequence set forth in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 108. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001264.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO:115, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:31, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:70, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:158, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:194, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:155, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:159, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:29, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 30. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO:36, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:58, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:170, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:190, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:176, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO:111, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO:112, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001265.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO:32, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 89, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 90, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001266.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 178, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 91, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 92. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001267.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, L-FR3 may comprise the amino acid sequence set forth in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 40, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 81, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 82. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001268.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 40, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 178, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 83, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 84. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001269.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 121, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:19, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 20, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 40, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 181, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 85, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 86, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001270.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 120, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:17, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO:18, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 40, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 181, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 85, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 86, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001271.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 57, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 121, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:19, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 20, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 89, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 90, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001272.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 57, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 69, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 25, and said CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 26. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 47, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 62, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 179, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 99, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 100, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001273.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 57, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 68, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 23, said CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 24. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 47, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 62, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 179, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 99, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 100, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001274.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 45, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 174, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 97, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 98, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001275.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 45, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 175, and H-FR4 may comprise the amino acid sequence set forth in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 93, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 94, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001276.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 119 and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 165, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:11, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 12. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 168, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 77, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 78, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001277.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 120, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 165, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:13, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO:14, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 168, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 77, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 78, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001278.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 118, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 165, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 9, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 10, or a pharmaceutically acceptable salt thereof. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 168, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 77, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 78, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001279.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 74, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 162, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:5, the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 6. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 51, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 63, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 182, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 183. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 103, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 104. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 001280.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO:115, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO:31, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO:70, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:158, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO:194, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO:155, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO:159, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:29, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 30. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO:34, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:58, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO:170, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:190, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:176, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 109, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 110. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000796.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 40, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 136, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 181, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO:186 to seq id no. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 85, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 86, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000802.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 57, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 67, said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO: 21, and said CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 22. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 47, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 62, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 179, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 99, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 100, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000803.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 116, and said L-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 166, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:15, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 16. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 45, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 173, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 95, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 96, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000804.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 117 and said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 165, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:7, and the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 8. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 42, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO:138, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 60, said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 168, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 177, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 185, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 77, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 78, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000805.

For example, an antigen binding protein described herein can comprise LCDR1-3 and L-FR 1-4. The LCDR1 can comprise SEQ ID NO: 113, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 56, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 74, said L-FR1 may comprise the amino acid sequence shown in SEQ ID NO: 157, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 191, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 162, and L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 160, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise VL and CL, and the VL may comprise SEQ ID NO:5, the CL may comprise the amino acid sequence set forth in SEQ ID NO:196, or a pharmaceutically acceptable salt thereof. The antigen binding protein may comprise an antibody light chain, which may comprise SEQ ID NO: 6. The antigen binding protein may also comprise HCDR1-3 and H-FR 1-4. Wherein the HCDR1 can comprise SEQ ID NO: 49, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 137, HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 63, and said H-FR1 may comprise the amino acid sequence set forth in SEQ ID NO: 169, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 188, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO: 182, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 183. The antigen binding protein may comprise a VH and a CH, and the VH may comprise SEQ ID NO: 101, and the CH may comprise the amino acid sequence shown in SEQ ID NO:195, or a pharmaceutically acceptable salt thereof. And the antigen binding protein may comprise an antibody heavy chain, which may comprise the amino acid sequence of SEQ ID NO: 102, or a pharmaceutically acceptable salt thereof. For example, the antigen binding protein comprises the same antibody light chain and antibody heavy chain as PR 000811.

Specific anti-CD 47 antibodiesAn isolated antigen binding protein as described herein that can compete with a particular anti-CD 47 antibody for binding to the proteinCD47, wherein the particular anti-CD 47 antibody may comprise a light chain variable region and a heavy chain variable region, the light chain variable region of the particular anti-CD 47 antibody may comprise LCDR1-3, the LCDR1-3 may comprise the amino acid sequence of SEQ ID NO: 139-141; the heavy chain variable region of the particular anti-CD 47 antibody may comprise HCDR1-3, and the HCDR1-3 may comprise the amino acid sequence of SEQ ID NO: 142 and 144. The light chain variable region amino acid sequence of the specific anti-CD 47 antibody can be SEQ ID NO: 153, and the heavy chain variable region of the specific anti-CD 47 antibody can be SEQ ID NO: 154.

in the present application, the LCDR1 of the particular anti-CD 47 antibody can comprise an amino acid sequence set forth in any one of: SEQ ID NO: 113. 114 and 115; the LCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 31. 56, 57, 122, 126 and 127; and the LCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 67-74, 116 and 121.

In the present application, the HCDR1 of the particular anti-CD 73 antibody may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51, and 53; the HCDR2 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 136-138; and the HCDR3 may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 58-63 and 55.

In the present application, the VL of the particular anti-CD 47 antibody may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29.

In the present application, the VH of the particular anti-CD 47 antibody may comprise an amino acid sequence set forth in any one of: SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111.

Reference in the present application to protein, polypeptide and/or amino acid sequences is also to be understood as including at least the following ranges: variants or homologues having the same or similar function as said protein or polypeptide. Variants having the same or similar function as the protein or polypeptide may be referred to as functional variants.

In the present application, the variant may be a protein or polypeptide having substitution, deletion or addition of one or more amino acids in the amino acid sequence of the protein and/or the polypeptide (e.g., an antibody or a fragment thereof specifically binding to CD47 protein). For example, the variant (e.g., functional variant) may comprise a protein or polypeptide that has been altered by at least 1, e.g., 1-30, 1-20, or 1-10, yet, e.g., 1, 2, 3, 4, or 5 amino acid substitutions, deletions, and/or insertions. The variant (e.g., functional variant) may substantially retain the biological properties of the protein or the polypeptide prior to the alteration (e.g., substitution, deletion, or addition). For example, the variant (e.g., functional variant) may retain at least 60%, 70%, 80%, 90%, or 100% of the biological activity (e.g., antigen binding capacity) of the protein or the polypeptide prior to alteration. For example, the substitution may be a conservative substitution.

In the present application, a portion of the amino acid sequence of the antigen binding protein may be homologous to a corresponding amino acid sequence in an antibody from a particular species, or belong to a particular class. For example, both the variable and constant regions of an antibody can be from the variable and constant regions of an antibody from one animal species (e.g., human). In the present application, the homolog may be a protein or polypeptide having at least about 85% (e.g., having at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology to the amino acid sequence of the protein and/or the polypeptide (e.g., an antibody or fragment thereof that specifically binds to CD47 protein).

In the present application, homology generally refers to similarity, similarity or relatedness between two or more sequences. The "percentage of sequence homology" can be calculated by: the two sequences to be aligned are compared in a comparison window, the number of positions in the two sequences at which the same nucleobase (e.g., A, T, C, G) or the same amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, gin, Cys, and Met) is determined to yield the number of matched positions, the number of matched positions is divided by the total number of positions in the comparison window (i.e., the window size), and the result is multiplied by 100 to yield the percentage of sequence homology. Alignment to determine percent sequence homology can be accomplished in a variety of ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. One skilled in the art can determine suitable parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the full length of the sequences being compared or over a region of the target sequence. The homology can also be determined by the following method: FASTA and BLAST. The FASTA algorithm is described in "improved tools for biological sequence comparison" by w.r.pearson and d.j.lipman, proceedings of the national academy of sciences of the united states (proc.natl.acad.sci.), 85: 2444 2448, 1988; and "rapid and sensitive protein similarity search" by d.j.lipman and w.r.pearson, Science, 227: 1435-1441, 1989. BLAST algorithms are described in "a basic local contrast (alignment) search tool" by s.altschul, w.gish, w.miller, e.w.myers and d.lipman, journal of molecular biology, 215: 403-410, 1990.

In the present application, the isolated antigen binding protein binds to human and monkey-derived CD47. The monkey may be a cynomolgus monkey. The binding affinity of an anti-CD 47 antibody to CD47 can be determined by any method known in the art. In some cases, binding affinity can be determined by competitive ELISA, RIA, or surface plasmon resonance such as BIAcore. In some specific cases, binding affinity is determined by surface plasmon resonance. For example, binding affinity and dissociation rate can be determined by BIAcore. In some cases, the KD for the isolated antigen binding protein to bind to CD47 can be 3x10^-9M is or below. In some particular cases, the KD for the isolated antigen binding protein to bind to CD47 can be 3x10^-9M、2x10^- ⁹M、1x10^-9M、1x10^-10M、1x10^-11M is or below. In some casesIn this form, the KD may be from about 1pM to about 500 pM. In other cases, the KD can be about 500pM to about 1 nM. In other cases, the KD can be about 1nM to about 10 nM.

In the present application, the isolated antigen binding protein can specifically bind to CD47. The CD47 described herein can be CD47 on the surface of a tumor cell. The tumor cells may be Jurkat cells or Raji cells. In some cases, the EC of the antigen binding protein binding to CD47 on Jurkat cells₅₀Values may be from about 10pM to about 10 nM. In other cases, the EC₅₀Values may be from about 100pM to about 100 nM. In other cases, EC₅₀Values may be from about 1pM to about 1 nM. In other cases, the EC₅₀Values may be from about 0.1nM to about 1nM, for example, in FACS measurements. For example, in a FACS assay, EC of the antigen binding protein binds to CD47 on Jurkat cells₅₀Values may be from about 10pM to about 10nM, e.g., from about 10pM to about 1nM, to about 1nM to about 10 nM. In the present application, the antigen binding protein does not bind or specifically bind to an antigen other than CD47, e.g., does not bind or specifically bind to SIPR α, CD147, PD-1, PD-L1, CTLA-4, CD80, CD86, CD28H, B7H3, B7H4, and ICOS.

The antigen binding proteins also bind to CD47 on the cell surface of cynomolgus monkey, e.g. to Peripheral Blood Mononuclear Cells (PBMCs) of cynomolgus monkeys. In some cases, the binding may be detected by FACS, and the resulting EC50 value may be about 0.02nM to about 0.2 nM. The EC50 measured by FACS may be in some cases about 0.01nM to about 0.3nM, about 0.01nM to about 0.5nM, or about 0.01nM to about 1.0 nM.

In the present application, the isolated antigen binding protein may inhibit the binding of CD47 to sirpa. In the present application, the CD47 may be the extracellular domain of human CD47. The extracellular domain of human CD47 described herein can comprise the sequence from position 19 to position 141 of the amino acid sequence shown in UniProt under accession number Q08722. For example, the isolated antigen binding protein inhibits the IC of human CD47 binding to SIRPa as measured by ELISA₅₀Values may be from about 0.5nM to about 5.0 nM. In some cases, the IC that blocks binding₅₀The value may be about1.0nM to about 3.0 nM. In other cases, it may be about 1.5nM to about 2.5 nM. For example, the isolated antigen binding protein may inhibit human CD47 binding to sirpa by greater than 90% (e.g., greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99% or more), as measured by ELISA.

In the present application, the isolated antigen binding protein may block the binding of CD47 on the surface of tumor cells to sirpa. The tumor cell described herein can be a Jurkat cell. The CD47 antigen binding proteins described herein are capable of blocking the binding of sirpa to human CD47. For example, the isolated antigen binding protein blocks the IC of CD47 binding to SIRPa on the surface of Jurkat cells₅₀Values may be from about 0.01nM to about 5.0nM, from about 0.02nM to about 3.0nM or from about 0.03nM to about 2.0nM. in some cases, the IC₅₀Values may be from about 0.1nM to about 1.0nM, as measured, for example, by FACS.

In the present application, the isolated antigen binding protein may not cause a coagulation response. In certain instances, the level of agglutination in the presence of the CD47 antigen binding proteins of the present application is reduced by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% compared to the level of agglutination in the presence of the existing CD47 antibodies, indicating that the CD47 antigen binding proteins of the present application do not elicit a coagulation response. For example, the antigen binding protein does not elicit a coagulation response at concentrations between about 065nM and about 665 nM.

In the present application, the isolated antigen binding protein can cause phagocytosis of tumor cells by macrophages. The tumor cells described herein can include Jurkat and OVCAR3 tumor cells. For example, the isolated antigen binding protein can cause phagocytosis of Jurkat by macrophages, as measured by flow cytometry, by labeling Jurkat cells with CFSE, and the phagocytosis index can be CFSE⁺CD14⁺Cell occupancy Total CD14⁺A percentage of cells, and the phagocytic index can be at least greater than 30% (e.g., at least greater than 40%, at least greater than 50%, at least greater than 60%, at least greater than 70%, at least greater than 80%),At least greater than 90% or more). EC that can cause phagocytosis of OVCAR3 by macrophages by the isolated antigen binding protein₅₀Values may be about 0.1nM to about 5.0nM (e.g., about 0.2nM to about 3.0nM, about 0.2nM to about 2.0nM, or about 0.5nM to about 1.5 nM). In some cases, for example, macrophages are labeled with CD14-APC and can be about 0.5nM to about 1.5nM as measured by flow cytometry.

The present application examines the ability of the isolated antigen binding protein to cause phagocytosis of human erythrocytes by macrophages. For example, the isolated antigen binding protein can have a phagocytosis index that can cause phagocytosis of human erythrocytes by macrophages, as measured by FACS, of less than 35% (e.g., can be less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, or less). The isolated antigen binding proteins described herein cause macrophages to phagocytose tumor cells more strongly than erythrocytes. The antigen binding proteins described herein are capable of inhibiting tumor growth and/or tumor cell proliferation. In certain instances, the antigen binding protein may slow tumor volume growth, eliminate tumor growth, or may slow the tendency for weight loss. This effect can be detected by establishing a mouse tumor model, for example, a Raji mouse model or an MC38 mouse model. Wherein the tumor comprises a solid tumor and/or a hematological tumor. Solid tumors may include ovarian cancer, etc., and hematological tumors may include leukemia, lymphoma. The leukemia may include Acute Lymphocytic Leukemia (ALL), Acute Myelogenous Leukemia (AML), Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML), and the like. Lymphomas may include hodgkin's lymphoma, indolent and aggressive non-hodgkin's lymphoma, burkitt's lymphoma, and the like.

Nucleic acids, vectors, cells and methods of preparation

In another aspect, the present application also provides isolated one or more nucleic acid molecules encoding the antibodies, antigen-binding fragments thereof, described herein. For example, each of the one or more nucleic acid molecules may encode the antibody, an antigen-binding fragment thereof in its entirety, or a portion thereof (e.g., one or more of HCDR1-3, LCDR1-3, VL, VH, light chain, or heavy chain).

The nucleic acid molecules described herein can be isolated. For example, it may be produced or synthesized by: (i) in vitro amplified, e.g., by Polymerase Chain Reaction (PCR), (ii) recombinantly produced by cloning, (iii) purified, e.g., by enzymatic cleavage and gel electrophoresis fractionation, or (iv) synthesized, e.g., by chemical synthesis. In certain embodiments, the isolated nucleic acid is a nucleic acid molecule prepared by recombinant DNA techniques.

In the present application, nucleic acids encoding the antibodies, antigen-binding fragments thereof, can be prepared by a variety of methods known in the art, including, but not limited to, overlap extension PCR using restriction fragment procedures or using synthetic oligonucleotides, as described in Sambrook et al, Molecular Cloning, Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; and Ausube et al Current Protocols in Molecular Biology, Greene Publishing and Wiley-Interscience, New York N.Y., 1993.

In another aspect, the present application provides one or more vectors comprising one or more of the nucleic acid molecules described herein. One or more of the nucleic acid molecules may be included in each vector. In addition, other genes may be included in the vector, such as marker genes that allow selection of the vector in an appropriate host cell and under appropriate conditions. In addition, the vector may contain expression control elements that allow for the proper expression of the coding region in an appropriate host. Such control elements are well known to those skilled in the art and may include, for example, promoters, ribosome binding sites, enhancers and other control elements that regulate gene transcription or mRNA translation, among others. In certain embodiments, the expression control sequence is a regulatable element. The specific structure of the expression control sequence may vary depending on the function of the species or cell type, but typically comprises 5 ' non-transcribed sequences and 5 ' and 3 ' non-translated sequences, such as TATA box, capping sequences, CAAT sequences, etc., which are involved in initiation of transcription and translation, respectively. For example, the 5' non-transcriptional expression control sequence may comprise a promoter region that may comprise a promoter sequence for a transcriptional control functional linkage nucleic acid. The expression control sequence may also include an enhancer sequence or an upstream activator sequence. In the present application, suitable promoters may include, for example, promoters for SP6, T3, and T7 polymerase, the human U6RNA promoter, the CMV promoter, and artificial hybrid promoters thereof (e.g., CMV), wherein a portion of the promoter may be fused to a portion of the promoter of other cellular proteins (e.g., human GAPDH, glyceraldehyde-3-phosphate dehydrogenase) genes, which may or may not contain additional introns. One or more of the nucleic acid molecules described herein can be operably linked to the expression control element.

The vector may include, for example, a plasmid, cosmid, virus, phage, or other vector commonly used in, for example, genetic engineering. For example, the vector is an expression vector.

In another aspect, the present application provides a host cell that may comprise one or more nucleic acid molecules described herein and/or one or more vectors described herein. In certain embodiments, each or each host cell may comprise one or more of the nucleic acid molecules or vectors described herein. In certain embodiments, each or each host cell may comprise a plurality (e.g., 2 or more) or a plurality (e.g., 2 or more) of the nucleic acid molecules or vectors described herein. For example, a vector described herein can be introduced into the host cell, e.g., a prokaryotic cell (e.g., a bacterial cell), a CHO cell, an NS/0 cell, an HEK293T cell, or an HEK293A cell, or other eukaryotic cell, such as a plant-derived cell, a fungal or yeast cell, and the like. The vectors described herein can be introduced into the host cell by methods known in the art, such as electroporation, lipofectine transfection, lipofectamine transfection, and the like. For example, the host cell may be COS, CHO, NSO, sf9, sf21, DH5a, BL21(DE3) or TG 1.

In another aspect, the present application provides methods of making the antibodies, antigen-binding fragments thereof, described herein. The method may comprise culturing a host cell described herein under conditions such that the antibody, antigen-binding fragment thereof, is expressed. For example, these methods can be performed by using an appropriate medium, an appropriate temperature, an appropriate incubation time, and the like, which are known to those of ordinary skill in the art.

In certain instances, the methods can further comprise the step of harvesting (e.g., isolating and/or purifying), the antibodies, antigen-binding fragments thereof, described herein. For example, protein G-Sepharose or protein A-Sepharose may be used for affinity chromatography, and the antibody, antigen-binding fragment thereof, described herein may be purified and isolated by gel electrophoresis and/or high performance liquid chromatography, or the like.

Pharmaceutical composition and application

In another aspect, the present application provides a pharmaceutical composition comprising an antigen-binding fragment or variant described herein, a nucleic acid molecule described herein, a vector described herein, a host cell described herein, and optionally a pharmaceutically acceptable carrier.

The pharmaceutically acceptable carrier generally refers to a carrier that can be used to prepare a pharmaceutical composition or formulation, is generally safe, non-toxic, and is neither biologically nor otherwise undesirable. The carrier used is generally one suitable for administration to humans or other mammals. In preparing the compositions, the active ingredient is generally mixed with, diluted by, or enclosed within a carrier. When the carrier serves as a diluent, it may be a solid, semi-solid or liquid material which acts as a vehicle or medium for the active ingredient of the antibody. The pharmaceutically acceptable carrier may include buffers, antioxidants, preservatives, low molecular weight polypeptides, proteins, hydrophilic polymers, amino acids, sugars, chelating agents, counter ions, metal complexes, and/or nonionic surfactants, and the like.

The pharmaceutical compositions may include an antigen binding protein as described herein, and may also include one or more active compounds in combination. The active compound may be a compound that does not adversely affect the antigen binding protein, or may be an agent that enhances its function, such as a cytotoxic agent, cytokine, chemotherapeutic agent or growth inhibitory agent. The compounds used in combination may be present in combination in an amount effective for the desired purpose.

In the present application, the pharmaceutical composition may be formulated for oral administration, intravenous administration, intramuscular administration, in situ administration at the tumor site, inhalation, rectal administration, vaginal administration, transdermal administration or administration via subcutaneous depot. Solutions or suspensions for transdermal administration or via subcutaneous depot may include the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetate, citrate or phosphate; and tonicity adjusting substances such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, for example hydrochloric acid or sodium hydroxide.

The pharmaceutical composition can be used for inhibiting tumor growth. For example, the pharmaceutical compositions of the present application may inhibit or delay the progression or progression of a disease, may reduce tumor size (even substantially eliminate tumors), and/or may alleviate and/or stabilize a disease state. For example, it can be expressed by the reduction of the body weight growth of the tumor mouse and the fluorescence signal intensity of the tumor cells.

The pharmaceutical compositions described herein may comprise a prophylactically and/or therapeutically effective amount of the antibody, antigen-binding fragment thereof. The prophylactically and/or therapeutically effective amount is the amount required to be able to prevent and/or treat (at least partially treat) a disease or disorder and/or any complications thereof in a subject suffering from or at risk of developing the same.

In another aspect, the application provides the use of the antibody, antigen-binding fragment thereof, in the manufacture of a medicament for the prevention or treatment of a disease or disorder.

In another aspect, the antibodies, antigen-binding fragments thereof, provided herein are useful for preventing or treating a disease or disorder. The prevention or treatment of a disease or disorder may refer to inhibiting or delaying the development or progression of a disease or disorder. For example, it can be used to inhibit the development or progression of tumors. For example, tumor growth or tumor cell proliferation can be inhibited.

In another aspect, the present application provides the antibody or antigen-binding fragment thereof for use in the prevention or treatment of a tumor.

In another aspect, the present application provides a method of inhibiting the binding of CD47 to sirpa comprising administering the antibody, antigen-binding fragment thereof, the nucleic acid molecule, the vector, the host cell, and/or the pharmaceutical composition described herein. For example, the method may be an in vitro or ex vivo method. For example, the method may be ELISA or FACS.

In another aspect, the present application provides a method of preventing or treating a tumor comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof, the molecular nucleic acid, the vector, the host cell, and/or the pharmaceutical composition described herein. In the present application, the tumor includes a solid tumor and/or a hematological tumor. For example, the tumor comprises lymphoma, leukemia, ovarian cancer.

Combination drug

The antigen binding proteins, pharmaceutical compositions, and methods of the present application can be administered in combination with other therapeutic agents and/or modalities. The combined administration generally refers to two (or more) different therapies being delivered to a subject during administration to the subject such that the therapeutic effects on the subject overlap at some point in time. In certain instances, delivery of the first therapy is still occurring when delivery of the second therapy is initiated, such that there is an overlap in administration. This is sometimes referred to herein as "simultaneous" or "parallel delivery". In other cases, delivery of one therapy ends before delivery of the other therapy begins. In some cases, the treatment is more effective due to the combined administration, and the effects of the two treatments may be partially additive, fully additive, or greater than additive.

In the present application, the antigen binding protein or pharmaceutical composition may be combined with the administration of one or more additional therapies, such as surgery, radiation therapy, or another therapeutic agent. In one embodiment, the additional therapy may include chemotherapy, e.g., a cytotoxic agent. In one embodiment, the additional therapy may comprise a targeted therapy, such as a tyrosine kinase inhibitor, a proteasome inhibitor, or a protease inhibitor. In one embodiment, the additional therapy may comprise: an anti-inflammatory compound, an anti-angiogenic compound, an anti-fibrotic compound, or an anti-proliferative compound, such as a steroid, a biological immunomodulator, a monoclonal antibody, an antibody fragment, an aptamer, a siRNA, an antisense molecule, a fusion protein, a cytokine receptor, a bronchodilator, a statin, an anti-inflammatory agent (e.g., methotrexate), or an NSAID. In another embodiment, the additional therapies may comprise different classes of combination therapies. The antigen binding protein, pharmaceutical composition or formulation and the additional therapy described herein may be administered simultaneously or sequentially.

In another aspect, the present application also provides the following embodiments:

1. an isolated antigen binding protein comprising an antibody light chain variable region VL comprising any one of LCDR1, LCDR2 and LCDR3 and an antibody heavy chain variable region VH comprising any one of HCDR1, HCDR2 and HCDR3, wherein the VL comprises the amino acid sequence of SEQ ID NO: 153, and VH comprises the amino acid sequence shown in SEQ ID NO: 154.

2. The isolated antigen binding protein of embodiment 1, wherein the LCDR1 comprises SEQ ID NO: 139, wherein LCDR1 preferably comprises the amino acid sequence set forth in SEQ ID NO: 113. 114 and 115, or a pharmaceutically acceptable salt thereof.

3. The isolated antigen binding protein of embodiment 1, wherein the LCDR2 comprises SEQ ID NO: 140, wherein LCDR2 preferably comprises the amino acid sequence set forth in SEQ ID NO: 31. 56, 57, 122, 126 and 127.

4. The isolated antigen binding protein of embodiment 1, wherein the LCDR3 comprises SEQ ID NO: 141, wherein LCDR3 preferably comprises the amino acid sequence set forth in SEQ ID NO: 67-74, 116-121.

5. The isolated antigen binding protein of embodiment 1, wherein the HCDR1 comprises SEQ ID NO: 142, wherein HCDR1 preferably comprises the amino acid sequence set forth in SEQ ID NO: 32. 34, 36, 38, 40, 42, 45, 47, 49, 51 and 53.

6. The isolated antigen binding protein of embodiment 1, wherein the HCDR2 comprises SEQ ID NO: 143, wherein HCDR2 preferably comprises the amino acid sequence set forth in SEQ ID NO: 136 and 138.

7. The isolated antigen binding protein of embodiment 1, wherein the HCDR3 comprises SEQ ID NO: 144, wherein HCDR3 preferably comprises the amino acid sequence set forth in SEQ ID NO: 55 and any one of 58-63.

8. The isolated antigen binding protein of any one of embodiment 1, wherein the VL comprises SEQ ID NO: 1.3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29.

9. The isolated antigen binding protein of any of embodiment 1, wherein the VH comprises SEQ ID NO: 75. 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109 and 111.

10. An isolated antigen binding protein as described in embodiment 1, comprising an antibody light chain LC, and the LC comprises the amino acid sequence of SEQ ID NO: 197, wherein said LC preferably comprises the amino acid sequence set forth in SEQ ID NO: 2. 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30.

11. An isolated antigen binding protein as described in embodiment 1 comprising an antibody heavy chain HC, and the HC comprises the amino acid sequence of SEQ ID NO: 198, wherein the HC preferably comprises the amino acid sequence shown in SEQ ID NO: 76. 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110 and 112.

12. The isolated antigen binding protein of embodiment 1, having one or more of the following properties:

1) capable of specifically binding human-and monkey-derived CD47, preferably with a KD ≦ 3x 10-9M;

2) can inhibit the binding of CD47 and SIRPa;

3) capable of binding human blood erythrocytes;

4) in the erythrocyte coagulation assay, no coagulation reaction is caused;

5) can cause phagocytosis of tumor cells by macrophages; and

6) can inhibit tumor growth and/or tumor cell proliferation.

13. One or more isolated nucleic acid molecules encoding the isolated antigen binding protein of any one of embodiments 1-12.

14. A vector comprising the nucleic acid molecule of embodiment 13.

15. A cell comprising a nucleic acid molecule according to embodiment 13 or a vector according to embodiment 14.

16. A method of making the isolated antigen binding protein of any one of embodiments 1-12, the method comprising culturing the cell of embodiment 15 under conditions such that the isolated antigen binding protein of any one of embodiments 1-12 is expressed.

17. A pharmaceutical composition comprising an isolated antigen binding protein of any one of embodiments 1-12, a nucleic acid molecule of embodiment 13, a vector of embodiment 14, and/or a cell of embodiment 15, and optionally a pharmaceutically acceptable carrier.

18. Use of an isolated antigen binding protein of any one of embodiments 1-12, a nucleic acid molecule of embodiment 13, a vector of embodiment 14, a cell of embodiment 15, and/or a pharmaceutical composition of embodiment 17 in the manufacture of a medicament for preventing, ameliorating, and/or treating a tumor.

19. A method of inhibiting CD47 binding to sirpa, the method comprising administering the isolated antigen binding protein of any one of embodiments 1-12.

Without intending to be bound by any theory, the following examples are merely intended to illustrate the fusion proteins, preparation methods, uses, etc. of the present application, and are not intended to limit the scope of the invention of the present application.

Examples

The examples shown below are intended to illustrate specific embodiments of the invention and are not intended to limit the scope of the specification or claims in any way. The examples do not include detailed descriptions of conventional methods, such as those used to construct vectors and plasmids, methods of inserting genes encoding proteins into such vectors and plasmids, or methods of introducing plasmids into host cells. The Laboratory Manual, 2nd edition, Cold spring Harbor Laboratory Press.

Example 1 mouse immunization and obtaining antibody molecules

Experimental animals, which may be mice, rats, rabbits, sheep, camels, etc., may be immunized with the CD47 antigen to obtain antibody molecules that specifically bind to CD47. Typically, the resulting antibody molecules are of non-human origin. After obtaining non-human antibodies, these molecules need to be humanized using antibody engineering techniques to reduce immunogenicity and improve druggability. However, the humanization process of antibodies has technical complexity, and molecules that are humanized and modified tend to have reduced affinity for antigens. On the other hand, advances in transgenic technology have made it possible to develop genetically engineered mice that carry the human immunoglobulin immune repertoire and have the endogenous murine immune repertoire deleted. The antibody generated by the transgenic mouse has a fully human sequence, so that further humanized modification is not needed, and the efficiency of developing a therapeutic antibody is greatly improved. The Harbour H2L2 mouse (Harbour Antibodies BV) is a transgenic mouse carrying a human immunoglobulin immune repertoire that produces Antibodies with intact human antibody variable domains and rat constant domains.

1.1 immunization of mice with the CD47 antigen

Harbour H2L2 mice were immunized in multiple rounds with soluble recombinant human CD47-Fc fusion protein. The antigenic protein is mixed with an immunological adjuvant to form an immunogen reagent, and then the immunogen is injected subcutaneously through the groin or is injected intraperitoneally. In each round of immunization, each mouse received a total injected dose of 100 microliters. In the first round of immunization, each mouse received an immunization with 50. mu.g of an immunogenic reagent formulated in a 1: 1 volume ratio of antigenic protein (human CD47-Fc) in complete Freund's adjuvant (Sigma # F5881). In each subsequent round of booster immunizations, each mouse received an immunization with an immunogen reagent formulated with 25 μ g of antigenic protein mixed with Sigma Adjuvant System Adjuvant (Sigma # S6322). The interval between each round of booster immunization is at least two weeks, usually not more than five rounds of booster immunization. The immunization time is 0, 14, 28, 42, 56 and 70 days; and at days 49, 77, mouse serum antibody titers were detected. 3 days before cell fusion, the last booster immunization was performed at a dose of 25. mu.g of antigen protein per mouse.

1.2 obtaining hybridoma monoclonal and antibody sequences

When the specific antibody titer of CD47 in the serum of a mouse reaches a certain level, taking out splenocytes of the mouse and fusing the splenocytes with a myeloma cell line to obtain hybridoma cells; after multiple rounds of screening and cloning of the hybridoma cells, 21 hybridomas expressing anti-CD 47 monoclonal antibody molecules were isolated. The isolated hybridoma cells and the monoclonal antibodies expressed by them are represented using the corresponding clone numbers: 11B8, 15D1, 18C4, 32F1, 35a11, 39a8, 40D7, 73F1, 74B6, 75F6, 78B6, 80B6, 81E5, 83G1, 84G2, 85G2, 86D7, 87E9, 87H6, 88G3, 89E 3. The isolated hybridomas express antibody molecules having intact human variable domains and heavy and light chains of rat constant domains. The monoclonal antibody is further identified, and a plurality of hybridoma clones are selected for sequencing according to the parameters of the monoclonal antibody, such as the binding capacity of the monoclonal antibody to human CD47, the binding capacity of the monoclonal antibody to cynomolgus monkey CD47, the binding capacity of the monoclonal antibody for inhibiting CD47 and SIRP alpha and the like. The nucleotide sequence encoding the variable domain of the antibody molecule and the corresponding amino acid sequence are obtained using conventional sequencing means of hybridomas. In this example, the sequence of the variable domain of the anti-CD 47 monoclonal antibody molecule obtained from immunized Harbour H2L2 mice was a human antibody sequence. The CDR sequences of antibody variable domains can be analyzed by Kabat or Chothia or other CDR definition rules. In this example, a rule called Combined definition combining the Kabat definition with the Chothia definition was also used for the analysis.

1.3 preparation of fully human recombinant antibodies

After obtaining the light and heavy chain variable domain sequences encoding the antibody molecules, the light and heavy chain variable domain sequences and the corresponding human antibody light and heavy chain constant domain sequences can be subjected to fusion expression by adopting the conventional recombinant DNA technology to obtain the recombinant antibody molecules. In this example, the antibody heavy chain variable domain sequence (VH) was genetically synthesized and cloned into a mammalian cell expression plasmid vector encoding the human IgG4 antibody heavy chain constant domain sequence to encode the full-length heavy chain that produced IgG4 antibody, and the S228P mutation (serine to proline at position 228 according to EU numbering) was introduced into the IgG4 heavy chain constant region to increase the stability of IgG4 antibody. Antibody light chain variable domain sequences (VL) were genetically synthesized and cloned into mammalian cell expression plasmid vectors encoding human antibody Ig kappa light chain constant domain sequences to encode full-length light chains that produce antibodies. In this example, since the sequence of the variable domain of the anti-CD 47 monoclonal antibody molecule obtained from immunized HarbourH2L2 mice was a human antibody sequence, this example also yielded fully human anti-CD 47 recombinant IgG4 antibody.

The plasmid coding the heavy chain of the antibody and the plasmid coding the light chain of the antibody are simultaneously transfected into a mammalian host cell (such as a human embryonic kidney cell HEK293), and the purified CD47 recombinant antibody with the correct pairing assembly of the light chain and the heavy chain can be obtained by utilizing the conventional recombinant protein expression and purification technology. Specifically, HEK293 cells were cultured in FreeStyle^TMF17 Expression Medium (Thermo # A1383504). Before transient transfection, cell concentration was adjusted to 6-8 x10⁵Cells/ml, cultured in a shaker at 37 ℃ for 24 hours with 8% CO2 at a cell concentration of 1.2X 10⁶Cells/ml. 30ml of cultured cells were prepared. The above plasmid encoding the heavy chain of the antibody and the plasmid encoding the light chain of the antibody were mixed in a ratio of 2: 3A total of 30. mu.g of plasmid was mixed and dissolved in 1.5ml of Opti-MEM reduced serum medium (Thermo, 31985088) and sterilized by filtration through a 0.22 μm filter. Further, 120. mu.l of 1mg/ml PEI (Polysciences, Inc #23966-2) was dissolved in 1.5ml of Opti-MEM and allowed to stand for 5 minutes. Slowly adding PEI into the plasmid, incubating for 10 min at room temperature, slowly dropping the mixed solution of the plasmid PEI while shaking the culture flask, and adding 8% CO at 37 deg.C₂Cultured in a shaker for 5 days. Cell viability was observed after 5 days. Collecting the culture, centrifuging at 3300g for 10 min, and collecting the supernatant; the supernatant was then centrifuged at high speed to remove impurities. Balancing the cells containing MabSelect with PBS (pH7.4)^TM(GE Healthcare Life Science #71-5020-91 AE) gravity column (Bio-Rad #7311550), 2-5 column volumes washed. Passing the supernatant sample through a column; the column was washed with 5-10 column volumes of PBS, followed by elution of the target protein with 0.1M glycine at pH3.5, followed by adjustment to neutrality with Tris-HCl pH 8.0, and finally by concentration of the exchange solution to PBS buffer using an ultrafiltration tube (Millipore, UFC901024) to obtain a purified CD47 antibody solution. Finally using NanoDrop (Thermo Scientific)^TMNanoDrop^TMOne) measuring the concentration, subpackaging and storing for later use.

1.4 sequences of fully human recombinant anti-CD 47 antibodies

Table 2 lists the amino acid sequences of the light and heavy chain variable domains, the full length amino acid sequence of the light chain, the full length amino acid sequence of the heavy chain (human IgG4), and the amino acid sequences of the CDRs defined according to the Chothia definition rules, of the CD47 antibody of this example.

Example 2 sequence analysis and expression purification of antibodies

The heavy chain variable domain sequence of the antibody is derived from events such as gene rearrangement of an embryonic line gene V, D, J gene segment of a heavy chain gene group on a chromosome, somatic high-frequency mutation and the like; the light chain variable domain sequence is derived from the events of gene rearrangement and somatic hypermutation of the germline gene V, J gene segment of the light chain gene group. Gene rearrangement and somatic hypermutation are major factors in increasing antibody diversity. Antibodies derived from the same germline V gene segment may also produce different sequences, but overall the similarity is higher. Using algorithms such as IMGT/DomainGapAlign (http:// IMGT. org/3Dstructure-DB/cgi/DomainGapAlign cgi) or NCBI/IgBLAST (https:// www.ncbi.nlm.nih gov/IgBLAST /) the germline gene segments that are likely to undergo gene rearrangement can be deduced from the variable domain sequences of the antibodies. The antibody sequences of example 1 and table 2 were analyzed and germline gene V gene segments for the heavy chain variable domain (VH) and light chain variable domain (VL) are listed in table 3.

Chemical modifications, known as post-translational modifications (PTMs), are sometimes introduced after the translational synthesis of a protein or polypeptide amino acid chain in a cell. For antibodies, the site of some PTMs is very conserved, e.g., the conserved amino acid asparagine Asn at position 297 (EU numbering) of the constant domain of the human IgG1 antibody is usually glycosylated to form a sugar chain whose structure is critical for antibody structure and associated effector functions. However, if PTMs are present in the variable domains, particularly the antigen binding regions (e.g., CDRs), of an antibody, then the presence of these PTMs may have a greater effect on antigen binding and may also have a change in the physicochemical properties of the antibody. For example, glycosylation, deamidation, isomerization, oxidation, and the like may all increase the instability or heterogeneity of antibody molecules, thereby increasing the difficulty and risk of antibody development. Avoiding some potential PTMs is therefore very important for the development of therapeutic antibodies. As experience has accumulated, it has been found that some PTMs are highly correlated with the composition of amino acid sequences, particularly the "pattern" of adjacent amino acid compositions, such that potential PTMs can be predicted from the primary amino acid sequence of the protein. For example, the sequence pattern of N-x-S/T (asparagine at the first position, any amino acid other than non-proline at the second position, and serine or threonine at the third position) predicts an N-linked glycosylation site. It is possible that the amino acid sequence pattern leading to PTM is derived from an germline gene sequence, e.g.the human germline gene segment IGHV3-33 naturally has a glycosylation pattern NST in the FR3 region; it may also be derived from somatic high-frequency mutations. Table 3 lists the predicted PTMs of the variable domains VH and VL of the antibody of example 1. Specifically, NSS or NLT may be a glycosylation site, and NS or NT or NN may be a deamidation site.

The amino acid sequence pattern of a PTM may be disrupted by amino acid mutation, thereby reducing or eliminating the formation of a particular PTM. There are different mutation design methods depending on the antibody sequence and PTM sequence pattern. One approach is to replace the "hot spot" amino acid (e.g., N or S in the NS pattern) with an amino acid with similar physicochemical properties (e.g., mutation of N to Q). If the PTM sequence pattern is derived from somatic high-frequency mutations and is not present in the germline gene sequence, another approach would be to replace the sequence pattern with the corresponding germline gene sequence. In practice, multiple mutation design methods may be used for the same PTM sequence pattern.

Table 4 lists the new antibody molecules (referred to as PTM variants) obtained by amino acid mutation of the sequences of six antibodies with potential PTM sites from example 1. Table 5 lists the light and heavy chain variable domain amino acid sequences, the full length light chain amino acid sequence, the full length heavy chain (human IgG4) amino acid sequence, and the amino acid sequences of the CDRs defined according to the Chothia definition rules for these PTM variants in this example. All designed PTM variants were purified recombinant antibodies as described in example 1.3 and further validated in subsequent functional experiments.

TABLE 2 screening of monoclonal anti-CD 47 hybridomas and recombinant antibodies thereof

TABLE 3 germline gene analysis and post-translational modification site (PTM) analysis of CD47 antibody sequences

TABLE 4 mutant site design of CD47 antibody sequences

TABLE 5 antibodies obtained after mutating PTM

Example 3 antigen binding proteins are able to bind to CD47 on the surface of human and monkey cells

3.1 ability to bind to CD47 on the surface of Jurkat and Raji target cells

Jurkat cells (ATCC # TIB-152) and Raji cells (ATCC # CCL-86) were separately cultured and expanded in 96-well plates (Corning #3799) at 1X 10 cells per well⁵Separately, 100. mu.l of the CD47 antibody obtained in example 1 or 2 and a control antibody (Tab1, Tab2, Tab3) were added at different concentrations, and the isotype hIgG4SP was used as a control. After mixing, incubation was carried out at 4 ℃ for 1 hour. 1% BSA (Amresco #0332-100G) was washed 3 times, 100. mu.l of 4. mu.g/mL goat anti-human IgG (H + L) -Alexa 488 was added, and incubated at 4 ℃ for 1 hour. After 3 washes, flow cytometry tests were performed (BD Biosciences # Canto II cytometer). Calculating half maximal binding Effect Concentration (EC) from the assay results₅₀) And with EC₅₀The values and the maximum MFI value were used as the relative binding activity evaluation basis. In the present application, Tab1, Tab2 and Tab3 are all from the prior art, Hu5F9-G4 of Forty Seven, CC-90002 of Celgene and SRF-231 of Surface Oncology, respectively.

The results are shown in FIG. 1 and tables 6-7 below. The CD47 antibodies obtained in example 1 or 2 both bound to Jurkat cells, had no significant difference in binding activity from Tab1 and Tab2 (FIGS. 1A-1B), and had better binding ability than Tab 3.

PR000806, PR000807, PR000808 and Tab1, Tab2 all bound to CD47 on the cell surface of Jurkat (FIG. 1B) and Raji (FIG. 1C), with no significant difference in binding activity and stronger affinity than Tab3 for Jurkat and Raji.

TABLE 6 binding of antigen binding proteins to Jurkat cell surface CD47

TABLE 7 binding of antigen binding proteins PR000806, PR000807, PR000808 to Jurkat and Raji surface CD47

3.2 binding to CD47 on the surface of cynomolgus PBMC cells

The binding ability of the CD47 antibody to cynomolgus PBMC was examined by the method of 3.1, and the results are shown in fig. 2 and table 8. The CD47 antibodies before PTM mutation obtained in example 1 can bind to CD47 on the surface of cynomolgus monkey PBMC cells, and EC₅₀The value is less than Tab 2.

TABLE 8 binding of antigen binding proteins to cynomolgus monkey surface CD47

Example 4 antigen binding proteins are capable of binding to human and cynomolgus monkey CD47 proteins

Human CD47(Sino Biological #12283-HCCH) and monkey CD47(Acro Biosystems # CD7-C52H1) were each diluted to 0.25. mu.g/ml with PBS and added to 96-well plates (Corning #9018) per minuteMu.l of the wells were incubated overnight at 4 ℃. After discarding the liquid, the plates were washed 3 times with PBST, blocked with 2% BSA and incubated at room temperature for 1 hour. The blocking solution was discarded and the plate was washed 5 times with PBST buffer (ph7.4, containing 0.05% tween-20) and the CD47 antibody obtained in example 1 or 2 and three CD47 antibodies of the prior art: tab1(Forty Seven, Hu5F9-G4), Tab2(Celgene, CC-90002), and Tab3(Surface aerology, SRF-231) were diluted 5-fold in sequence from 66.7nM concentration, and 10 concentration gradients were added to 100. mu.l/well and incubated at 37 ℃ for 1 hour, using hIgG4SP (Crown Bio # C0045-4) as a control. After washing 3 times, a 4000-fold diluted secondary goat anti-human HRP antibody (Invitrogen # A18805) was added and incubated at 37 ℃ for 1 hour. After washing, 100. mu.l/well of TMB (Beyotime # P0209) was added and left for 5 minutes at room temperature in the dark; the reaction was stopped by adding 100. mu.l/well of a stop solution (BBI life sciences # E661006-0200) to each well, and absorbance (OD450) at 450nm was measured in a microplate reader (PerkinElemer # Enspire). Calculating half maximal binding Effect Concentration (EC) from the assay results₅₀) And with EC₅₀The values and the maximum OD values were used as the basis for evaluation of relative binding activity.

The results are shown in FIG. 3 and tables 9-10 below. Fig. 3A and table 9 show that the CD47 antibodies before PTM mutation obtained in example 1 all bound to cynomolgus monkey-derived CD47 protein. Figures 3B-3C and table 10 show the activity of PR00806, PR000807, PR000808 in combination with human (figure 3B) and monkey (figure 3C) CD47. PR00806, PR000807, PR000808, Tab1, Tab2 and Tab3 can be combined with human CD47 protein, and their combination activity has no significant difference, and the EC of PR00806, PR000807 and PR000808 can be used for preventing and curing various diseases of human body₅₀Below 0.03nM, the binding capacity is better than that of Tab 3. PR00806, PR000807, PR000808, Tab1 and Tab2 can be combined with monkey CD47 protein, and the combination activity is not obviously different, but the combination ability of Tab3 and monkey CD47 protein is weaker.

TABLE 9 binding of antigen binding proteins to monkey CD47 protein

TABLE 10 binding of antigen binding proteins to human or monkey CD47 protein

Example 5 antigen binding proteins are able to inhibit the binding of human CD47 protein to SIRP alpha

Human CD47-ECD-hFc (Sino Biological #12283-H02H) was diluted to 1. mu.g/ml with PBS and added to a 96-well plate (Corning #9018) at 100. mu.l per well and coated overnight at 4 ℃. After 3 washes, blocking was performed with 100. mu.l of 2% BSA at room temperature for 1 hour. The CD47 antibody obtained in example 1 or 2 and control antibodies (Tab1, Tab2, Tab3) were diluted with PBST to different concentration gradients, mixed with 1. mu.g/ml histidine-tagged SIRP α protein (Sino Biological #11612-H08H), added to the well plate, and incubated with hIgG4 as a control for 1 hour at room temperature. After washing 3 times, 100. mu.l of histidine-tagged HRP antibody (Biolegend #652504) was added to each well and reacted at room temperature for 1 hour. After washing for 3 times, 100. mu.l of TMB developing solution was added, and color development was carried out for 5 minutes at room temperature in the dark. The reaction was stopped by adding stop solution, and absorbance (OD450) at 450nm was measured by a microplate reader (PerkinElemer # Enspire). With IC₅₀The value, the minimum OD value and the maximum inhibition rate are used as evaluation criteria of relative inhibition activity.

The results are shown in FIG. 4 and tables 11-12 below. FIG. 4A and Table 11 show that the CD47 antibodies obtained in example 1 all inhibited the binding of human CD47 to SIRPa. FIG. 4B and Table 12 show that PR00806, PR000807 and PR000808 can inhibit the combination of human CD47 and SIRPa, the inhibition rate is greater than 97%, and the inhibition activity is not significantly different from that of Tab1, Tab2 and Tab 3.

TABLE 11 antigen binding proteins inhibit the binding of human CD47 to SIRP alpha

TABLE 12 antigen binding proteins inhibit the binding of human CD47 to SIRP alpha

Example 6 antigen binding proteins are able to block the binding of CD47 on Jurkat cells to SIRP alpha

After the human Jurkat cells highly expressing CD47 were expanded and cultured, the cells were cultured at 1X 10 cells per well⁵One was added to a 96-well plate. The CD47 antibody obtained in example 1 or 2 and control antibodies (Tab1, Tab2, Tab3) were diluted with PBS to different concentration gradients, mixed with 1. mu.g/ml histidine-tagged SIRPa protein, added to each well, isotype hIgG4SP as a control, and incubated at 4 ℃ for 1 hour. 1% BSA was washed 2 times, added with a 100. mu.l volume of a secondary antibody Biotin-conjugated anti-His Tag antibody (GenScript # A00613), and incubated at 4 ℃ for 1 hour. After centrifugation, the cells were washed 2 times with 1% BSA, and then subjected to flow cytometric assay after 3 times by adding a volume of 100. mu.l of tri-anti-PE-Conjugated streptavidin (Affymetrix # DC). With IC₅₀The value, the minimum MFI value and the maximum inhibition rate are used as evaluation basis of relative blocking activity.

The results are shown in FIG. 5 and tables 13-14 below. FIG. 5A and Table 13 show that the CD47 antibodies obtained in example 1 both blocked the binding of CD47 on the cell surface of Jurkat to SIRPa. FIG. 5B and Table 14 show that PR00806, PR000807 and PR000808 can block the binding of CD47 and SIRPa on the surface of Jurkat cells, and the blocking activity is not significantly different from that of Tab1 and Tab2, and IC is₅₀Are all less than 0.7nM, whereas Tab3 has a weaker blocking capacity.

TABLE 13 antigen binding proteins block binding of CD47 on Jurkat cells to SIRP alpha

TABLE 14 antigen binding proteins block binding of CD47 on Jurkat cells to SIRP alpha

Example 7 antigen binding proteins do not cause agglutination of erythrocytes

The CD47 antibodies obtained in example 1 or 2 were analyzed for the presence or absence of an agglutination reaction of erythrocytes. Human erythrocytes were diluted to 4% with PBS and added to a 96-well plate (Corning #3799), and then the test antibody and the control antibody (Tab1, Tab2, Tab3) were diluted 2-fold from 200nM, and the same type hIgG4SP was used as a control, and incubated at 37 ℃ for 1 hour. The effect of the antibody on agglutination of human erythrocytes was evaluated by comparison with the sharpness of the red spot edge in wells without antibody.

FIGS. 6A-6B show that, in comparison with wells without antibody, other antigen binding proteins did not cause agglutination except for PR00796, with sharp red spots; tab1 caused agglutination reaction, and the red spots were blurred and dispersed.

Example 8 detection of the binding Capacity of antigen binding proteins to erythrocytes

The activity of binding of the CD47 antibody obtained in example 1 or 2 to erythrocytes was examined. Human erythrocytes were diluted in PBS and added to a 96-well plate (Corning #3799) at 1X 10 per well⁵Then, 100. mu.l of anti-CD 47 antibody and control antibody (Tab1, Tab2, Tab3) at different concentrations were added thereto, and the isotype hIgG4SP was used as a control, and the mixture was mixed and incubated at 4 ℃ for 1 hour. 1% BSA (Amresco #0332-100G) was washed 3 times, added 100. mu.l of 4. mu.g/mL goat anti-human IgG (H + L) -Alexa 488, incubated at 4 ℃ for 1 hour, and subjected to flow cytometric assay after washing 3 times (BD Biosciences # Canto II cytometer). By EC₅₀The value and the maximum MFI value were used as the evaluation criteria of the relative binding activity.

The ability of antigen binding proteins to bind to erythrocytes is shown in FIG. 7 and tables 15-16 below.

TABLE 15 antigen binding proteins binding to erythrocytes

Comparison of antigen binding proteins binding to Jurkat cells (example 3)Ability to bind to Red Blood Cells (RBC), EC of antigen binding protein binding to tumor cells and red blood cells was calculated₅₀And the maximum MFI, the results are given in table 16 below. As seen from the results, the antigen-binding protein obtained in example 1 or 2 had a greater ability to bind to Jurkat than to red blood cells, as compared to the control antibody.

TABLE 16 ratio of antigen binding proteins to tumor cell and erythrocyte binding Capacity

Example 9 antigen binding proteins are able to induce phagocytosis of tumor cells by macrophages

9.1 causing phagocytosis of Jurkat cells by macrophages

CD14 isolated from human Peripheral Blood Mononuclear Cells (PBMC)⁺The mononuclear cells (Miltenyi Biotec #130-&D Systems #216-MC/CF) for 7 days to differentiate into macrophages, trypsinized, and plated on 96-well plates at 5X 10 per well⁴Simultaneously, Jurkat cells labeled with 0.25. mu.M carboxyfluorescein succinimidyl ester (CFSE) (Life-technology # C34570) -were added at a ratio of 1: 1, and then diluted anti-CD 47 antibody and control antibody (Tab1, Tab2, Tab3) were added, mixed and incubated at 37 ℃ for 2 hours. Unphagocytosed Jurkat cells were washed away with 1% BSA and the direct antibody CD14-APC (BD Pharmingen #561708) was added. Flow cytometry (ACEA # Novocyte) was used to evaluate phagocytosis, which is CFSE⁺CD14⁺Cell occupancy Total CD14⁺Percentage of cells.

The results are shown in FIG. 8 and tables 17-19 below. FIG. 8A and Table 17 show that the antibodies CD47 obtained in example 1 before PTM mutation both induced phagocytosis of Jurkat cells by macrophages, and the ability to induce phagocytosis was superior to that of Tab 2. FIG. 8B and Table 18 show that PR000806, PR000807 and PR000808 all induced phagocytosis of Jurkat cells by macrophages with a greater capacity to induce phagocytosis than Tab2 and Tab 3. Table 19 shows that the CD47 antibody obtained in example 1 or 2 caused phagocytosis of Jurkat cells by macrophages at various concentrations.

TABLE 17 antigen binding proteins cause phagocytosis of Jurkat cells by macrophages

TABLE 18 antigen binding proteins cause phagocytosis of Jurkat cells by macrophages

TABLE 19 antigen binding proteins cause phagocytosis of Jurkat cells by macrophages

9.2 causing phagocytosis of OVCAR3 cells by macrophages

Phagocytosis of OVCAR3(ATCC # HTB-161) cells by macrophages was examined as described in example 9.1, EC was determined by flow cytometry₅₀And phagocytosis index.

As shown in fig. 9 and table 20 below, PR00806, PR000807, and PR000808 all caused phagocytosis of OVCAR3 cells by macrophages with no significant difference; EC (EC)₅₀The values and maximum phagocytic index show that PR00806, PR000807, PR000808 induce phagocytosis more strongly than Tab 3.

TABLE 20 antigen binding proteins cause phagocytosis of OVCAR3 cells by macrophages

Example 10 detection of the ability of an antigen binding protein to cause phagocytosis of human erythrocytes by macrophages

Example 9 test example1 or 2, inducing phagocytosis of human erythrocytes by macrophages, and measuring EC by flow cytometry₅₀And phagocytosis index.

The results are shown in FIG. 10 and tables 21-22 below. Fig. 10 and table 21 show the ability of CD47 antibody to cause phagocytosis of human erythrocytes by macrophages after PTM removal. Table 22 shows the ability of the CD47 antibody obtained in example 1 or 2 to induce phagocytosis of hRBC by macrophages at various concentrations. The results show that the antigen binding proteins described herein (e.g., CD47 antibodies) are effective in causing phagocytosis of hrbcs by macrophages with similar or higher efficacy than control antibodies.

TABLE 21 antigen binding proteins cause phagocytosis of human erythrocytes by macrophages

TABLE 22 antigen binding proteins cause phagocytosis of human erythrocytes by macrophages

Example 11 in vivo anti-tumor Activity of antigen binding proteins

11.1 in vivo anti-tumor Activity of antigen binding proteins on NCG-Raji mouse model

The antitumor activity of the antigen-binding proteins was evaluated in the NCG-Raji mouse model. 1X 10 of Raji cells⁷Each 0.1mL was inoculated into the tail vein of NCG mice. On the third day after cell inoculation, the mice were randomly divided into 10 groups of 8 mice each, and dosing was started. Each antibody to be tested was injected intraperitoneally at 0.3mg/kg, 1mg/kg and 3mg/kg, 10. mu.l/g, 1 dose every 3 days, 6 doses consecutively, with hIgG4 of the same type as a control. Mouse body weight and tumor volume were recorded.

Table 23 shows the tumor inhibition ratio (TGI) of each antibody injected compared to isotype IgG4, which is dose dependent. FIGS. 11A-11B show the tumor volume and body weight trend in mice injected with PR000806, PR000808 and Tab 1. The tumor volume growth of the group mice injected with PR000806, PR000808 slowed down. The results show that PR000806 and PR000808 showed in vivo inhibition of tumor growth, and PR000808 showed better effect than Tab1 at 3mg/kg and 1mg/kg doses.

TABLE 23 antigen binding proteins inhibit tumor growth

11.2 in vivo anti-tumor Activity of antigen binding proteins on B-hSIRPa/hCD47-MC38-hCD47 mouse model

The anti-tumor activity of antigen binding proteins was evaluated in a B-hSIRPa/hCD47-MC38-hCD47 mouse model. 1 × 10 MC38 cells stably expressing human CD47 protein⁶One/0.1 mL was inoculated subcutaneously into the right side of B-hSIRPA/hCD47 mice. After cell inoculation, when the mean tumor volume reached approximately 80-120mm3, the mice were randomized into 7 groups of 8 mice each, and dosing was initiated. Each antibody to be tested was injected intraperitoneally at 5mg/kg and 10mg/kg, 10. mu.l/g, 1 time every 3 days, 6 times consecutively, with hIgG4 isotype as control. Mouse body weight and tumor volume were recorded.

FIGS. 12A-12B show the tumor volume and body weight trend in mice injected with PR000806, PR001265 and Tab 1. The tumor volume growth of PR000806, PR001265 injected group mice slowed. The results show that PR000806 and PR001265 were able to inhibit tumor growth in vivo, with PR000806 and PR001265 having better tumor inhibition than Tab1 at high doses.

Example 12 toxicity testing of antigen binding proteins

The mice homozygous for B-hSIRPA/hCD47 were randomly divided into groups of 4 mice per group for 8 groups, and were administered by intraperitoneal injection at a dose of 10mpk on day 8 after division, with the first administration being on day 0 of treatment for 1 time (day 0). Body weights of mice were measured 1 time per day during dosing and observation, and the measurements were recorded. Blood was collected 7 days before administration (normal value), 1 day after treatment, and 7 days after treatment, and subjected to routine blood analysis. The results of the mouse body weight change and blood routine analysis are shown in FIGS. 13-15.

The experiment was terminated 11 days after dosing, at the end of the experiment, the animals were euthanized and the mice were photographed.

FIG. 13 shows the body weight change of mice before and after treatment. With the exception of homo-type hIgG4, mice lost weight following treatment with other antigen-binding proteins or antibodies, with the degree of weight loss following treatment with Tab1, PR000806, PR000808 and PR001281 being greater than the other three molecules, consistent with the in vitro binding of the antibody molecules to the antigen on erythrocytes, but at approximately 7 days post-treatment, the mice began to return to normal levels.

FIGS. 14-16 show the results of routine blood tests on day 1 and day 7 of the mouse model before and after treatment. As can be seen from the images, on the first day after administration, some parameters in the blood routine of the sample group were changed from those in the control group hIgG4SP, but after 7 days after administration, the indexes were all recovered to normal as the body's compensation function was exerted.

Finally, from body weight and blood routine data, none of the antibody molecules PR000806, PR000808 and PR001281, PR001265, PR001275 and PR001276 targeting CD47 showed unrecoverable hematologic toxicity.

Example 13 affinity test (Biacore) of the ability of antigen binding proteins to bind to CD47

13.1 coupling of Protein A

Protein A (Thermo Fisher #21181) was coupled to 4 channels of CM5 using 1 XHBS-EP + (GEHealthcare # BR-1006-69) as running buffer at a flow rate of 10 ul/min: 1) setting the injection time to 800s, mixing 50mM NHS and 200mM EDC in a volume ratio of 1: 1 and injecting the mixture into 4 channels; 2) diluting Protein A to 20 μ g/ml with sodium acetate of pH4.5, injecting for 800 s; 3) 1M ethanolamine 800s is injected to block the residual active carboxyl on the surface of the chip. After blocking, the instrument was equilibrated for two hours with 1 XHBS-EP + buffer and the final amount of Protein A coupled was about 2000 RU.

13.2 kinetic testing

A multi-cycle kinetic pattern is set up, each cycle comprising capture of the antibody, binding of the analyte and regeneration of the chip. Each antibody was diluted to 1. mu.g/ml, injected into 2, 3, 4 channels 40s at a flow rate of 10. mu.l/min, and captured by the pre-coupled Protein A in an amount of about 200 RU. Human CD47(Sino biological #12283-HCCH) and cynomolgus monkey CD47(Acro Biosystems # CD7-C52H1) were injected into the four channels in succession at a concentration gradient of 0nM, 1.25nM, 2.5nM, 5nM, 10nM, 20nM, 40nM, at a flow rate of 30ul/min, with an injection time of 180s and a dissociation time of 900s, respectively. Finally, glycine (10mM, pH1.5) was injected at the same flow rate for 30s to regenerate the chip.

13.3 data analysis

The experimental results were analyzed using Biacore T200 analysis software 2.0, with channel 1 subtracted as the reference channel and the analytical model selected as a 1: 1 kinetic fitting model.

The results are shown in Table 24 below. The experimental results show that 5 antibodies have good binding with human CD47 and cynomolgus monkey CD47.

TABLE 24 binding affinities of antigen binding proteins to cynomolgus monkey and human CD47

Example 14 antigen binding proteins bind to CD47 with high specificity

Histidine-tagged human CD47(Sino Biological #12283-HCCH), SIPR α (Sino Biological #11612-H08H), CD147(Sino Biological #10186-H08H), PD-1(Novoprotein # CD91), PD-L1(Novoprotein # C315), CTLA-4(Sino Biological #11159-H08H), CD80(Sino Biological #10698-H08H), CD86(Sino Biological #10699-H08H), CD H (Acrosystem # CDH-H52H3), B7H3(Sino #11188-H08H), B7H4(Sino Biological #10738-H H) and Acrosystem (Acrosystem # PBS # 96H 6) were incubated with PBS # 96 ml/96 ml, 100 μ g/ml and PBS # 96/ml. After discarding the liquid, the plates were washed 3 times with PBST, blocked with 2% BSA and incubated at room temperature for 1 hour. The blocking solution was discarded, and the plate was washed 5 times with PBST buffer (pH7.4, containing 0.05% Tween-20), the CD47 antibodies PR001265, PR001275, PR001276 and Tab1(Forty Seven, Hu5F9-G4) obtained in example 1 or 2 were diluted to 10nM and 100nM, 100. mu.l/well was added, and incubation was carried out at 37 ℃ for 1 hour with the same type of hIgG4SP (Crown Bio # C0045-4) as a control. After washing 3 times, a 4000-fold diluted secondary goat anti-human HRP antibody (Invitrogen # A18805) was added and incubated at 37 ℃ for 1 hour. After washing, 100. mu.l/well of TMB (Beyotime # P0209) was added and left for 5 minutes at room temperature in the dark; the reaction was stopped by adding 100. mu.l/well of a stop solution (BBI life sciences # E661006-0200) to each well, and absorbance values at 450nm and 570nm were measured in a microplate reader (PerkinElemer # Enspire). The OD450/570 ratio was calculated.

The results show (figure 17) that the antigen binding proteins of the present application bind CD47 with high specificity and do not bind the non-CD 47 antigens SIPR α, CD147, PD-1, PD-L1, CTLA-4, CD80, CD86, CD28H, B7H3, B7H4 and ICOS.

The foregoing detailed description is provided by way of illustration and example, and is not intended to limit the scope of the appended claims. Various modifications of the presently recited embodiments will be apparent to those of ordinary skill in the art and are intended to be within the scope of the appended claims and their equivalents.

Sequence listing

<110> and platinum medicine (Shanghai) Limited liability company

<120> anti-CD 47 antigen binding protein and application thereof

<130> 0113-PA-012CN

<160> 198

<170> PatentIn version 3.5

<210> 1

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000808 light chain variable region

<400> 1

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 2

<211> 214

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000808 light chain

<400> 2

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 3

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000807 light chain variable region

<400> 3

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Val Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 4

<211> 214

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000807 light chain

<400> 4

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Val Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 5

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000811, PR001280 and PR001281 light chain variable region

<400> 5

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Ser Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 6

<211> 214

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000811, PR001280 and PR001281 light chain

<400> 6

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Ser Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 7

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000805 light chain variable region

<400> 7

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Asn Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 8

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000805 light chain

<400> 8

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Asn Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 9

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001279 light chain variable region

<400> 9

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Asn Thr Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 10

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001279 light chain

<400> 10

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Asn Thr Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 11

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001277 light chain variable region

<400> 11

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Gln Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 12

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001277 light chain

<400> 12

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Gln Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 13

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001278 light chain variable region

<400> 13

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 14

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001278 light chain

<400> 14

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Ser Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 15

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802、PR000804、PR001266、PR001267、PR001268、PR001269、PR001

275. PR001276 light chain variable region

<400> 15

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Ser Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 16

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802、PR000804、PR001266、PR001267、PR001268、PR001269、PR001

275. PR001276 light chain

<400> 16

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Ser Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 17

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001271 light chain variable region

<400> 17

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 18

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001271 light chain

<400> 18

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Asn Thr Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 19

<211> 108

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001270, PR001272 light chain variable region

<400> 19

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Gln Ser Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 20

<211> 215

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001270, PR001272 light chain

<400> 20

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Arg Gln Tyr Gln Ser Tyr Ser Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

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

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 21

<211> 106

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803 light chain variable region

<400> 21

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Asn Ser Tyr Tyr Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 22

<211> 213

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803 light chain

<400> 22

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Asn Ser Tyr Tyr Thr

85 90 95

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

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 23

<211> 106

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001274 light chain variable region

<400> 23

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Asn Thr Tyr Tyr Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 24

<211> 213

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001274 light chain

<400> 24

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Asn Thr Tyr Tyr Thr

85 90 95

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

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 25

<211> 106

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001273 light chain variable region

<400> 25

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Gln Ser Tyr Tyr Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 26

<211> 213

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001273 light chain

<400> 26

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Gln Ser Tyr Tyr Thr

85 90 95

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

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 27

<211> 106

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000806 light chain variable region

<400> 27

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Arg Ser Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ile Asn Tyr Tyr Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 28

<211> 213

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000806 light chain

<400> 28

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Arg Ser Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ile Asn Tyr Tyr Thr

85 90 95

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

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 29

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000796, PR001264, PR001265 variable region in light chain

<400> 29

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

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

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Leu Tyr Asn Tyr Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 30

<211> 214

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000796, PR001264, PR001265 light chain

<400> 30

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

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

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Leu Tyr Asn Tyr Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 31

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR2 of PR000796, PR001264, PR001265 (Kabat, Chothia, Combined)

<400> 31

Gly Ala Ser Ile Arg Ala Thr

1 5

<210> 32

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001264 (Chothia)

<400> 32

Gly Asp Ser Val Asp Thr Tyr

1 5

<210> 33

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001264 (Combined)

<400> 33

Gly Asp Ser Val Asp Thr Tyr Tyr Trp Thr

1 5 10

<210> 34

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000796 (Chothia)

<400> 34

Gly Asp Ser Val Asn Thr Tyr

1 5

<210> 35

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000796 (Combined)

<400> 35

Gly Asp Ser Val Asn Thr Tyr Tyr Trp Thr

1 5 10

<210> 36

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001265 (Chothia)

<400> 36

Gly Asp Ser Val Gln Thr Tyr

1 5

<210> 37

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1(Combined) of PR001265

<400> 37

Gly Asp Ser Val Gln Thr Tyr Tyr Trp Thr

1 5 10

<210> 38

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000808 (Chothia)

<400> 38

Gly Gly Ser Ile Asp Asn Tyr

1 5

<210> 39

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000808 (Combined)

<400> 39

Gly Gly Ser Ile Asp Asn Tyr Tyr Trp Ser

1 5 10

<210> 40

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR001268，

PR001269，PR001270，

HCDR (Chothia) of PR001271

<400> 40

Gly Gly Ser Ile Asn Asn Tyr

1 5

<210> 41

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR001268，

PR001269，PR001270，

HCDR (Combined) of PR001271

<400> 41

Gly Gly Ser Ile Asn Asn Tyr Tyr Trp Ser

1 5 10

<210> 42

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000805，PR000806，PR000807，PR001266，PR001267，PR001272，PR001

277, PR001278, HCDR (Chothia) of PR001279

<400> 42

Gly Gly Ser Ile Ser Asn Tyr

1 5

<210> 43

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000805，PR000806，PR001266，PR001267，PR001272，PR001277，PR001

278, PR001279 HCDR1(Combined)

<400> 43

Gly Gly Ser Ile Ser Asn Tyr Tyr Trp Ser

1 5 10

<210> 44

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1(Combined) of PR000807

<400> 44

Gly Gly Ser Ile Ser Asn Tyr Tyr Trp Thr

1 5 10

<210> 45

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000804, PR001275, HCDR1 of PR001276 (Chothia)

<400> 45

Gly Gly Ser Ile Ser Ser Tyr

1 5

<210> 46

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000804, PR001275, HCDR1 of PR001276 (Combined)

<400> 46

Gly Gly Ser Ile Ser Ser Tyr Tyr Trp Ser

1 5 10

<210> 47

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, HCDR1 of PR001274 (Chothia)

<400> 47

Gly Gly Ser Ile Ser Thr Tyr

1 5

<210> 48

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, HCDR1 of PR001274 (Combined)

<400> 48

Gly Gly Ser Ile Ser Thr Tyr Tyr Trp Ser

1 5 10

<210> 49

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> 49

<400> 49

Gly Gly Ser Leu Asn Asn Tyr

1 5

<210> 50

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1(Combined) of PR000811

<400> 50

Gly Gly Ser Leu Asn Asn Tyr Tyr Trp Ser

1 5 10

<210> 51

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001280 (Chothia)

<400> 51

Gly Gly Ser Leu Gln Asn Tyr

1 5

<210> 52

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001280 (Combined)

<400> 52

Gly Gly Ser Leu Gln Asn Tyr Tyr Trp Ser

1 5 10

<210> 53

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001281 (Chothia)

<400> 53

Gly Gly Ser Leu Ser Asn Tyr

1 5

<210> 54

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR001281 (Combined)

<400> 54

Gly Gly Ser Leu Ser Asn Tyr Tyr Trp Ser

1 5 10

<210> 55

<211> 6

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR3 of PR000808 (Kabat, Chothia, Combined)

<400> 55

Gly Arg Trp Gly Ser Tyr

1 5

<210> 56

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR000804，PR000805，PR000811，PR001266，PR001267，PR001

268，PR001269，PR001270，PR001271，PR001272，PR001275，PR001276，

PR001277, PR001278, PR001279, PR001280, LCDR2 of PR001281 (Kabat, C)

hothia，Combined）

<400> 56

Lys Ala Ser Ser Leu Glu Ser

1 5

<210> 57

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, LCDR2 of PR001274 (Chothia, Kabat, Combined)

<400> 57

Lys Ala Ser Thr Leu Glu Ser

1 5

<210> 58

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000796, PR001264, HCDR3 of PR001265 (Chothia, Kabat, Combined)

<400> 58

Lys Lys Leu Ile Phe Asp Tyr

1 5

<210> 59

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR3 of PR000806 (Chothia, Kabat, Combined)

<400> 59

Lys Arg Ala Met Tyr Asp Tyr

1 5

<210> 60

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR000804，PR000805，PR001266，PR001267，PR001268，PR001

269，PR001270，PR001271，PR001272，PR001275，PR001276，PR001277，

PR001278, HCDR3 of PR001279 (Kabat, Chothia, Combined)

<400> 60

Lys Arg Gly Val Leu Asp Tyr

1 5

<210> 61

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR3 of PR000807 (Kabat, Chothia, Combined)

<400> 61

Lys Arg Thr Ala Phe Asp Tyr

1 5

<210> 62

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, HCDR3 of PR001274 (Chothia, Kabat, Combined)

<400> 62

Lys Arg Thr Val His Asp Tyr

1 5

<210> 63

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR3 of PR000811, PR001280, PR001281 (Chothia, Kabat, Combined)

<400> 63

Lys Arg Trp Gly Phe Asp Tyr

1 5

<210> 64

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR000808 (Kabat, Combined)

<400> 64

Asn Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 65

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR000805，PR000806，PR000808，PR000811，PR001266，PR001

267，PR001268，PR001269，PR001270，PR001271，PR001272，PR001277，

PR001278, PR001279, PR001280, HCDR1 of PR001281 (Kabat)

<400> 65

Asn Tyr Tyr Trp Ser

1 5

<210> 66

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000807 (Kabat)

<400> 66

Asn Tyr Tyr Trp Thr

1 5

<210> 67

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000803 (Kabat, Chothia, Combined)

<400> 67

Gln His Tyr Asn Ser Tyr Tyr Thr

1 5

<210> 68

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR001274 (Kabat, Chothia, Combined)

<400> 68

Gln His Tyr Asn Thr Tyr Tyr Thr

1 5

<210> 69

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR001273 (Kabat, Chothia, Combined)

<400> 69

Gln His Tyr Gln Ser Tyr Tyr Thr

1 5

<210> 70

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000796, PR001264, PR001265 (Chothia, Kabat, Combined)

<400> 70

Gln Leu Tyr Asn Tyr Trp Pro Leu Thr

1 5

<210> 71

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000807 (Kabat, Chothia, Combined)

<400> 71

Gln Gln Ser Tyr Ser Thr Pro Arg Thr

1 5

<210> 72

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000808 (Kabat, Chothia, Combined)

<400> 72

Gln Gln Ser Tyr Ser Thr Pro Tyr Thr

1 5

<210> 73

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000806 (Kabat, Chothia, Combined)

<400> 73

Gln Gln Tyr Ile Asn Tyr Tyr Thr

1 5

<210> 74

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000811, PR001280, PR001281 (Kabat, Chothia, Combined)

<400> 74

Gln Gln Tyr Asn Ser Tyr Ser Tyr Thr

1 5

<210> 75

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000807

<400> 75

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Met Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Thr Ala Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 76

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000807

<400> 76

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Met Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Thr Ala Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 77

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000805, PR001277, PR001278, PR001279

<400> 77

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 78

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000805, PR001277, PR001278, PR001279

<400> 78

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 79

<211> 114

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000808

<400> 79

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asp Asn Tyr

20 25 30

Tyr Trp Ser Trp Phe Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Ser Ser

<210> 80

<211> 441

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000808

<400> 80

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asp Asn Tyr

20 25 30

Tyr Trp Ser Trp Phe Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys

130 135 140

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu

145 150 155 160

Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu

165 170 175

Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr

180 185 190

Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val

195 200 205

Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro

210 215 220

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

225 230 235 240

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

245 250 255

Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr

260 265 270

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

275 280 285

Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

290 295 300

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

305 310 315 320

Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

325 330 335

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met

340 345 350

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

355 360 365

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

370 375 380

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

385 390 395 400

Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val

405 410 415

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

420 425 430

Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 81

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001268

<400> 81

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 82

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001268

<400> 82

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 83

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001269

<400> 83

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 84

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001269

<400> 84

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 85

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802, PR001270, PR001271

<400> 85

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 86

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000802, PR001270, PR001271

<400> 86

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 87

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000806

<400> 87

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

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

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Ala Met Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 88

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000806

<400> 88

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

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

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

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

85 90 95

Arg Lys Arg Ala Met Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 89

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001266, PR001272 heavy chain variable region

<400> 89

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 90

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001266, PR001272

<400> 90

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 91

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001267

<400> 91

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 92

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001267

<400> 92

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Gly Val Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 93

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001276

<400> 93

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Thr

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 94

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001276

<400> 94

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Thr

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 95

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000804

<400> 95

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Ser Ser Leu Thr

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 96

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000804

<400> 96

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Ser Ser Leu Thr

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 97

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001275

<400> 97

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

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

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 98

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001275

<400> 98

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

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

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 99

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, PR001274 heavy chain variable region

<400> 99

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Thr Val His Asp Tyr Trp Gly Gln Gly Ile Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 100

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000803, PR001273, PR001274

<400> 100

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

1 5 10 15

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

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Lys Arg Thr Val His Asp Tyr Trp Gly Gln Gly Ile Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 101

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000811

<400> 101

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 102

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000811

<400> 102

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Asn Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 103

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001280

<400> 103

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Gln Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 104

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001280

<400> 104

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Gln Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 105

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001281

<400> 105

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

Val Ser Ser

115

<210> 106

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001281

<400> 106

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Leu Ser Asn Tyr

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 107

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001264

<400> 107

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Val Asp Thr Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 108

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001264

<400> 108

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Val Asp Thr Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 109

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR000796

<400> 109

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

1 5 10 15

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

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 110

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR000796

<400> 110

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

1 5 10 15

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

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 111

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain variable region of PR001265

<400> 111

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Val Gln Thr Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 112

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain of PR001265

<400> 112

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Val Gln Thr Tyr

20 25 30

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

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

65 70 75 80

Lys Leu Ser Ser Val Thr Ser Ala Asp Thr Ala Met Tyr Tyr Cys Ala

85 90 95

Arg Lys Lys Leu Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 113

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802-806，PR000811，

LCDR1 of PR001266-1281 (Kabat, Chothia, Combined)

<400> 113

Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala

1 5 10

<210> 114

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR1 of PR000807, PR000808 (Kabat, Chothia, Combined)

<400> 114

Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu Asn

1 5 10

<210> 115

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR1 of PR000796, PR001264, PR001265 (Kabat, Chothia, Combined)

<400> 115

Arg Ala Ser Gln Ser Val Ser Ser His Leu Ala

1 5 10

<210> 116

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000802, PR000804, PR001266-1269, PR001275-1276 (Kabat, C)

hothia，Combined）

<400> 116

Arg Gln Tyr Asn Ser Tyr Ser Pro Tyr Thr

1 5 10

<210> 117

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR000805 (Kabat, Chothia, Combined)

<400> 117

Arg Gln Tyr Asn Thr Asn Ser Pro Tyr Thr

1 5 10

<210> 118

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR001279 (Kabat, Chothia, Combined)

<400> 118

Arg Gln Tyr Asn Thr Asn Thr Pro Tyr Thr

1 5 10

<210> 119

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR001277 (Kabat, Chothia, Combined)

<400> 119

Arg Gln Tyr Asn Thr Gln Ser Pro Tyr Thr

1 5 10

<210> 120

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR3 of PR001271, PR001278 (Kabat, Chothia, Combined)

<400> 120

Arg Gln Tyr Asn Thr Tyr Ser Pro Tyr Thr

1 5 10

<210> 121

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001270, LCDR3 of PR001272 (Kabat, Chothia, Combined)

<400> 121

Arg Gln Tyr Gln Ser Tyr Ser Pro Tyr Thr

1 5 10

<210> 122

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR2 of PR000806 (Kabat, Chothia, Combined)

<400> 122

Arg Ser Ser Ser Leu Asp Ser

1 5

<210> 123

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000804, PR001275, PR001276 (Kabat)

<400> 123

Ser Tyr Tyr Trp Ser

1 5

<210> 124

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803, PR001273, HCDR1 of PR001274 (Kabat)

<400> 124

Thr Tyr Tyr Trp Ser

1 5

<210> 125

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR1 of PR000796, PR001264, PR001265 (Kabat)

<400> 125

Thr Tyr Tyr Trp Thr

1 5

<210> 126

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR2 of PR000808 (Kabat, Chothia, Combined)

<400> 126

Val Ala Ser Arg Leu Gln Ser

1 5

<210> 127

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> LCDR2 of PR000807 (Kabat, Chothia, Combined)

<400> 127

Val Ala Ser Ser Leu Gln Ser

1 5

<210> 128

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001266, PR001268, HCDR2 of PR001272 (Kabat, Combined)

<400> 128

Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 129

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802, PR001267, PR001269, PR001270, HCDR2 of PR001271 (Kabat, Combi)

ned）

<400> 129

Tyr Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Ser Ser Leu Lys Ser

1 5 10 15

<210> 130

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR000803, PR000811, PR001273, PR001274, PR001280, PR001281 (Kab)

at，Combined）

<400> 130

Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 131

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR000807 (Kabat, Combined)

<400> 131

Tyr Ile Tyr Tyr Ser Gly Thr Thr Asp Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 132

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR0007963,PR000805,PR000806,PR001264,PR001265,PR001277,PR001278,P

HCDR2 of R001279 (Kabat, Combined)

<400> 132

Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 133

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR001276 (Kabat, Combined)

<400> 133

Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Pro Ser Leu Thr Ser

1 5 10 15

<210> 134

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR000804 (Kabat, Combined)

<400> 134

Tyr Ile Tyr Tyr Ser Gly Thr Thr Asn Tyr Asn Ser Ser Leu Thr Ser

1 5 10 15

<210> 135

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2 of PR001275 (Kabat, Combined)

<400> 135

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

1 5 10 15

<210> 136

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802, PR001266, PR001267, PR001268, PR001269, PR001270 and PR001

271 HCDR2(Chothia)

<400> 136

Tyr Tyr Ser Gly Asn

1 5

<210> 137

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2(Chothia) of PR000803, PR000811, PR001273, PR001274 and PR001281

<400> 137

Tyr Tyr Ser Gly Ser

1 5

<210> 138

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> HCDR2(Chothia) of PR000796, PR000804-808, 1264, 1265, 1275-

<400> 138

Tyr Tyr Ser Gly Thr

1 5

<210> 139

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of LCDR1

<220>

<221> X

<222> (6)..(6)

<223> X = I or V

<220>

<221> X

<222> (9)..(9)

<223> X = W, Y or H

<220>

<221> X

<222> (11)..(11)

<223> X = A or N

<400> 139

Arg Ala Ser Gln Ser Xaa Ser Ser Xaa Leu Xaa

1 5 10

<210> 140

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of LCDR2

<220>

<221> X

<222> (1)..(1)

<223> X = K, R, V or G

<220>

<221> X

<222> (2)..(2)

<223> X2= A or S

<220>

<221> X

<222> (4)..(4)

<223> X3= S, T, R or I

<220>

<221> X

<222> (5)..(5)

<223> X4= L or R

<220>

<221> X

<222> (6)..(6)

<223> X = E, D, Q or A

<220>

<221> X

<222> (7)..(7)

<223> X = S or T

<400> 140

Xaa Xaa Ser Xaa Xaa Xaa Xaa

1 5

<210> 141

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of LCDR3

<220>

<221> X

<222> (1)..(1)

<223> X = Q or R

<220>

<221> X

<222> (2)..(2)

<223> X = Q, L or H

<220>

<221> X

<222> (3)..(3)

<223> X = Y or-

<220>

<221> X

<222> (4)..(4)

<223> X = N, I, Q or-

<220>

<221> X

<222> (5)..(5)

<223> X = S, Y, T or N

<220>

<221> X

<222> (6)..(6)

<223> X = Y, W, N or Q

<220>

<221> X

<222> (7)..(7)

<223> X = S, T or-

<220>

<221> X

<222> (8)..(8)

<223> X = T or-

<220>

<221> X

<222> (9)..(9)

<223> X = P or-

<220>

<221> X

<222> (10)..(10)

<223> X = Y, R or L

<400> 141

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Thr

1 5 10

<210> 142

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> formula of HCDR1

<220>

<221> X

<222> (2)..(2)

<223> X = G or D

<220>

<221> X

<222> (4)..(4)

<223> X = I, L or V

<220>

<221> X

<222> (5)..(5)

<223> X = S, N, D or Q

<220>

<221> X

<222> (6)..(6)

<223> X = N, S or

<220>

<221> X

<222> (6)..(6)

<223> X = N, S or T

<400> 142

Gly Xaa Ser Xaa Xaa Xaa Tyr

1 5

<210> 143

<211> 5

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> formula of HCDR2

<220>

<221> X

<222> (5)..(5)

<223> X = T, N or S

<400> 143

Tyr Tyr Ser Gly Xaa

1 5

<210> 144

<211> 7

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> formula of HCDR3

<220>

<221> X

<222> (1)..(1)

<223> X = K or G

<220>

<221> X

<222> (2)..(2)

<223> X = R or K

<220>

<221> X

<222> (3)..(3)

<223> X = G, A, T, W or L

<220>

<221> X

<222> (4)..(4)

<223> X = V, M, G, I or A

<220>

<221> X

<222> (5)..(5)

<223> X = L, Y, H, F or-

<220>

<221> X

<222> (6)..(6)

<223> X = D or S

<400> 144

Xaa Xaa Xaa Xaa Xaa Xaa Tyr

1 5

<210> 145

<211> 25

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of H-FR1

<220>

<221> X

<222> (12)..(12)

<223> X = V or M

<220>

<221> X

<222> (13)..(13)

<223> X = K or Q

<220>

<221> X

<222> (23)..(23)

<223> X = T or S

<400> 145

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Xaa Val Ser

20 25

<210> 146

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of H-FR2

<220>

<221> X

<222> (3)..(3)

<223> X = S or T

<220>

<221> X

<222> (5)..(5)

<223> X = I or F

<220>

<221> X

<222> (8)..(8)

<223> X = P or A

<220>

<221> X

<222> (11)..(11)

<223> X = K or R

<220>

<221> X

<222> (18)..(18)

<223> X = Y or N

<400> 146

Tyr Trp Xaa Trp Xaa Arg Gln Xaa Pro Gly Xaa Gly Leu Glu Trp Ile

1 5 10 15

Gly Xaa Ile

<210> 147

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of H-FR3

<220>

<221> X

<222> (2)..(2)

<223> X = N or D

<220>

<221> X

<222> (4)..(4)

<223> X = N or Q

<220>

<221> X

<222> (5)..(5)

<223> X = P or S

<220>

<221> X

<222> (8)..(8)

<223> X = K or T

<220>

<221> X

<222> (12)..(12)

<223> X = T or S

<220>

<221> X

<222> (13)..(13)

<223> X = I or M

<220>

<221> X

<222> (15)..(15)

<223> X = V or I

<220>

<221> X

<222> (17)..(17)

<223> X = T or I

<220>

<221> X

<222> (25)..(25)

<223> X = K or N

<220>

<221> X

<222> (27)..(27)

<223> X = S, T or N

<220>

<221> X

<222> (30)..(30)

<223> X = T or I

<220>

<221> X

<222> (31)..(31)

<223> X = A or S

<220>

<221> X

<222> (32)..(32)

<223> X = A or G

<220>

<221> X

<222> (36)..(36)

<223> X = V or M

<220>

<221> X

<222> (40)..(40)

<223> X = A or G

<400> 147

Thr Xaa Tyr Xaa Xaa Ser Leu Xaa Ser Arg Val Xaa Xaa Ser Xaa Asp

1 5 10 15

Xaa Ser Lys Asn Gln Phe Ser Leu Xaa Leu Xaa Ser Val Xaa Xaa Xaa

20 25 30

Asp Thr Ala Xaa Tyr Tyr Cys Xaa Arg

35 40

<210> 148

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of H-FR4

<220>

<221> X

<222> (1)..(1)

<223> X = W or S

<220>

<221> X

<222> (5)..(5)

<223> X = T or I

<400> 148

Xaa Gly Gln Gly Xaa Leu Val Thr Val Ser Ser

1 5 10

<210> 149

<211> 23

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of L-FR1

<220>

<221> X

<222> (1)..(1)

<223> X = D or E

<220>

<221> X

<222> (3)..(3)

<223> X = Q or V

<220>

<221> X

<222> (9)..(9)

<223> X = S or A

<220>

<221> X

<222> (10)..(10)

<223> X = T or S

<220>

<221> X

<222> (13)..(13)

<223> X = A or V

<220>

<221> X

<222> (15)..(15)

<223> X = V or P

<220>

<221> X

<222> (17)..(17)

<223> X = D or E

<220>

<221> X

<222> (19)..(19)

<223> X = V or A

<220>

<221> X

<222> (21)..(21)

<223> X = I or L

<220>

<221> X

<222> (22)..(22)

<223> X = T or S

<400> 149

Xaa Ile Xaa Met Thr Gln Ser Pro Xaa Xaa Leu Ser Xaa Ser Xaa Gly

1 5 10 15

Xaa Arg Xaa Thr Xaa Xaa Cys

20

<210> 150

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of L-FR2

<220>

<221> X

<222> (8)..(8)

<223> X = K, N or Q

<220>

<221> X

<222> (11)..(11)

<223> X = K or R

<400> 150

Trp Tyr Gln Gln Lys Pro Gly Xaa Ala Pro Xaa Leu Leu Ile Tyr

1 5 10 15

<210> 151

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of L-FR3

<220>

<221> X

<222> (1)..(1)

<223> X = G or D

<220>

<221> X

<222> (2)..(2)

<223> X = V or I

<220>

<221> X

<222> (4)..(4)

<223> X = S or V

<220>

<221> X

<222> (14)..(14)

<223> X = E or D

<220>

<221> X

<222> (16)..(16)

<223> X = T or S

<220>

<221> X

<222> (19)..(19)

<223> X = I or V

<220>

<221> X

<222> (24)..(24)

<223> X = P or S

<220>

<221> X

<222> (25)..(25)

<223> X = D or E

<220>

<221> X

<222> (29)..(29)

<223> X = T or I

<400> 151

Xaa Xaa Pro Xaa Arg Phe Ser Gly Ser Gly Ser Gly Thr Xaa Phe Xaa

1 5 10 15

Leu Thr Xaa Ser Ser Leu Gln Xaa Xaa Asp Phe Ala Xaa Tyr Tyr Cys

20 25 30

<210> 152

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of L-FR4

<220>

<221> X

<222> (3)..(3)

<223> X = Q or G

<220>

<221> X

<222> (7)..(7)

<223> X = L or V

<400> 152

Phe Gly Xaa Gly Thr Lys Xaa Glu Ile Lys

1 5 10

<210> 153

<211> 109

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of light chain variable region

<220>

<221> X

<222> (1)..(1)

<223> X = D or E

<220>

<221> X

<222> (3)..(3)

<223> X = Q or V

<220>

<221> X

<222> (9)..(9)

<223> X = S or A

<220>

<221> X

<222> (10)..(10)

<223> X = T or S

<220>

<221> X

<222> (13)..(13)

<223> X = A or V

<220>

<221> X

<222> (15)..(15)

<223> X = V or P

<220>

<221> X

<222> (17)..(17)

<223> X = D or E

<220>

<221> X

<222> (19)..(19)

<223> X = V or A

<220>

<221> X

<222> (21)..(21)

<223> X = I or L

<220>

<221> X

<222> (22)..(22)

<223> X = T or S

<220>

<221> X

<222> (29)..(29)

<223> X = I or V

<220>

<221> X

<222> (32)..(32)

<223> X = W, Y or H

<220>

<221> X

<222> (34)..(34)

<223> X = a or N;

<220>

<221> X

<222> (42)..(42)

<223> X = K, N or Q

<220>

<221> X

<222> (45)..(45)

<223> X = K or R

<220>

<221> X

<222> (50)..(50)

<223> X = K, R, V or G

<220>

<221> X

<222> (51)..(51)

<223> X = A or S

<220>

<221> X

<222> (53)..(53)

<223> X = S, T, R or I

<220>

<221> X

<222> (54)..(54)

<223> X = L or R

<220>

<221> X

<222> (55)..(55)

<223> X = E, D, Q or A

<220>

<221> X

<222> (56)..(56)

<223> X21= S or T

<220>

<221> X

<222> (57)..(57)

<223> X = G or D

<220>

<221> X

<222> (58)..(58)

<223> X = V or I

<220>

<221> X

<222> (60)..(60)

<223> X = S or V

<220>

<221> X

<222> (70)..(70)

<223> X = E or D

<220>

<221> X

<222> (72)..(72)

<223> X = T or S

<220>

<221> X

<222> (75)..(75)

<223> X = I or V

<220>

<221> X

<222> (80)..(80)

<223> X = P or S

<220>

<221> X

<222> (81)..(81)

<223> X = D or E

<220>

<221> X

<222> (85)..(85)

<223> X = T or I

<220>

<221> X

<222> (89)..(89)

<223> X = Q or R

<220>

<221> X

<222> (90)..(90)

<223> X = Q, L or H

<220>

<221> X

<222> (91)..(91)

<223> X = Y or-

<220>

<221> X

<222> (92)..(92)

<223> X = N, I, Q or-

<220>

<221> X

<222> (93)..(93)

<223> X = S, T or N

<220>

<221> X

<222> (94)..(94)

<223> X = Y, W, N or Q

<220>

<221> X

<222> (95)..(95)

<223> X = S, T or-

<220>

<221> X

<222> (96)..(96)

<223> X = T or-

<220>

<221> X

<222> (97)..(97)

<223> X = P or-

<220>

<221> X

<222> (98)..(98)

<223> X = Y, R or L

<220>

<221> X

<222> (102)..(102)

<223> X = Q or G

<220>

<221> X

<222> (106)..(106)

<223> X = L or V

<400> 153

Xaa Ile Xaa Met Thr Gln Ser Pro Xaa Xaa Leu Ser Xaa Ser Xaa Gly

1 5 10 15

Xaa Arg Xaa Thr Xaa Xaa Cys Arg Ala Ser Gln Ser Xaa Ser Ser Xaa

20 25 30

Leu Xaa Trp Tyr Gln Gln Lys Pro Gly Xaa Ala Pro Xaa Leu Leu Ile

35 40 45

Tyr Xaa Xaa Ser Xaa Xaa Xaa Xaa Xaa Xaa Pro Xaa Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Xaa Phe Xaa Leu Thr Xaa Ser Ser Leu Gln Xaa

65 70 75 80

Xaa Asp Phe Ala Xaa Tyr Tyr Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

85 90 95

Xaa Xaa Thr Phe Gly Xaa Gly Thr Lys Xaa Glu Ile Lys

100 105

<210> 154

<211> 115

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> general formula of heavy chain variable region

<220>

<221> X

<222> (12)..(12)

<223> X = V or M

<220>

<221> X

<222> (13)..(13)

<223> X = K or Q

<220>

<221> X

<222> (23)..(23)

<223> X = T or S

<220>

<221> X

<222> (27)..(27)

<223> X = G or D

<220>

<221> X

<222> (29)..(29)

<223> X = I, L or V

<220>

<221> X

<222> (30)..(30)

<223> X = S, N, D or Q

<220>

<221> X

<222> (31)..(31)

<223> X = N, S or T

<220>

<221> X

<222> (35)..(35)

<223> X = S or T

<220>

<221> X

<222> (37)..(37)

<223> X = I or F

<220>

<221> X

<222> (40)..(40)

<223> X = P or A

<220>

<221> X

<222> (43)..(43)

<223> X = K or R

<220>

<221> X

<222> (50)..(50)

<223> X = Y or N

<220>

<221> X

<222> (56)..(56)

<223> X = T, N or S

<220>

<221> X

<222> (58)..(58)

<223> X = N or D

<220>

<221> X

<222> (60)..(60)

<223> X = N or Q

<220>

<221> X

<222> (61)..(61)

<223> X = P or S

<220>

<221> X

<222> (64)..(64)

<223> X = K or T

<220>

<221> X

<222> (68)..(68)

<223> X = T or S

<220>

<221> X

<222> (69)..(69)

<223> X = I or M

<220>

<221> X

<222> (71)..(71)

<223> X = V or I

<220>

<221> X

<222> (73)..(73)

<223> X = T or I

<220>

<221> X

<222> (81)..(81)

<223> X = K or N

<220>

<221> X

<222> (83)..(83)

<223> X = S, T or N

<220>

<221> X

<222> (86)..(86)

<223> X = T or I

<220>

<221> X

<222> (87)..(87)

<223> X = A or S

<220>

<221> X

<222> (88)..(88)

<223> X = A or G

<220>

<221> X

<222> (92)..(92)

<223> X = V or M

<220>

<221> X

<222> (96)..(96)

<223> X = A or G

<220>

<221> X

<222> (98)..(98)

<223> X = K or G

<220>

<221> X

<222> (99)..(99)

<223> X = R or K

<220>

<221> X

<222> (100)..(100)

<223> X = G, A, T, W or L

<220>

<221> X

<222> (101)..(101)

<223> X = V, M, G, I or A

<220>

<221> X

<222> (102)..(102)

<223> X = L, Y, H, F or-

<220>

<221> X

<222> (103)..(103)

<223> X = D or S

<220>

<221> X

<222> (105)..(105)

<223> X = W or S

<220>

<221> X

<222> (109)..(109)

<223> X = T or I

<400> 154

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Xaa Val Ser Gly Xaa Ser Xaa Xaa Xaa Tyr

20 25 30

Tyr Trp Xaa Trp Xaa Arg Gln Xaa Pro Gly Xaa Gly Leu Glu Trp Ile

35 40 45

Gly Xaa Ile Tyr Tyr Ser Gly Xaa Thr Xaa Tyr Xaa Xaa Ser Leu Xaa

50 55 60

Ser Arg Val Xaa Xaa Ser Xaa Asp Xaa Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Xaa Leu Xaa Ser Val Xaa Xaa Xaa Asp Thr Ala Xaa Tyr Tyr Cys Xaa

85 90 95

Arg Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Gly Gln Gly Xaa Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 155

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR3 of PR000796, PR001264, 1265 (Combined)

<400> 155

Asp Ile Pro Val Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys

20 25 30

<210> 156

<211> 23

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR1 of PR000807, PR000808

<400> 156

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys

20

<210> 157

<211> 23

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR 1(Combined) of PR000802-806, 811 and 1266-

<400> 157

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys

20

<210> 158

<211> 23

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR1 of PR000796, 1264 and 1265

<400> 158

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys

20

<210> 159

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR4 of PR000796, 1264 and 1265

<400> 159

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 160

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR4 of PR000802-806, 808, 811 and 1266-

<400> 160

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 161

<211> 10

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR4 (Combined) of PR000807

<400> 161

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 162

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR3 of PR000811, 1280 and 1281

<400> 162

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 163

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR3 of PR000808

<400> 163

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 164

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR3(Combined) of PR000807

<400> 164

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

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

20 25 30

<210> 165

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR1 of PR000805, PR001277, PR001278, PR001279

<400> 165

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ser

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 166

<211> 32

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR000803，PR000804，PR000806，PR001266，PR001267，PR001

268，PR001269，PR001270，PR001271，PR001272，PR001275，PR001276，

PR001273, L-FR3 of PR001274

<400> 166

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 167

<211> 25

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR1 of PR000807

<400> 167

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Met Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser

20 25

<210> 168

<211> 25

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR1 of PR000805, PR001277, PR001278, PR001279

<400> 168

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Ser

20 25

<210> 169

<211> 25

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000802，PR000803，PR000804，PR000806，PR000808，PR000811，PR001

266-

<400> 169

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser

20 25

<210> 170

<211> 25

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR1 of PR000796, PR001264, PR001265

<400> 170

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser

20 25

<210> 171

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR3 of PR000807

<400> 171

Thr Asp Tyr Asn Pro Ser Leu Lys Ser Arg Val Ser Ile Ser Val Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Asn Ser Val Thr Ala Gly

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 172

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR3 of PR000806

<400> 172

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Ile Asp

1 5 10 15

Ile Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Ile Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 173

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000806 H-FR3

<400> 173

Thr Asn Tyr Asn Ser Ser Leu Thr Ser Arg Val Thr Ile Ser Ile Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 174

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001275 H-FR3

<400> 174

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

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 175

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001276 H-FR3

<400> 175

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

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 176

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000796，PR001264，PR001265 H-FR3

<400> 176

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Ile Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ser Ala

20 25 30

Asp Thr Ala Met Tyr Tyr Cys Ala Arg

35 40

<210> 177

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR3 of PR000805, PR001266, PR001268, PR001272, PR001277-1279

<400> 177

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 178

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR001269，PR001267 H-FR3

<400> 178

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

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 179

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803，PR001273，PR001274 H-FR3

<400> 179

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Thr Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 180

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000808 H-FR3

<400> 180

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Thr Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Gly Arg

35 40

<210> 181

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR3 of PR000802, PR001270-1271

<400> 181

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

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Asn Leu Thr Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 182

<211> 41

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR3 of PR000811, PR001280-1281

<400> 182

Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser Val Asp

1 5 10 15

Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala

20 25 30

Asp Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 183

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR4 of PR000811, PR001280-1281

<400> 183

Ser Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 184

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR2 of PR000808

<400> 184

Tyr Trp Ser Trp Phe Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

1 5 10 15

Gly Asn Ile

<210> 185

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> PR000803，PR000804，PR000805，PR001275，PR001276，PR001273，PR001

274, PR001277, PR001278, H-FR4 of PR001279

<400> 185

Trp Gly Gln Gly Ile Leu Val Thr Val Ser Ser

1 5 10

<210> 186

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR4 of PR000796, PR000802, PR000806-808, PR0001264-1272

<400> 186

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 187

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR2 of PR000806

<400> 187

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

1 5 10 15

Gly Tyr Ile

<210> 188

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR2 of PR000802-805, PR000811, PR001266-1281

<400> 188

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

1 5 10 15

Gly Tyr Ile

<210> 189

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR2 of PR000807

<400> 189

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

1 5 10 15

Gly Tyr Ile

<210> 190

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> H-FR2 of PR000796, PR001264, PR001265

<400> 190

Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

1 5 10 15

Gly Tyr Ile

<210> 191

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR2 of PR000802-805, PR000808, PR000811, PR001266-1281

<400> 191

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 192

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR2 of PR000807

<400> 192

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile Tyr

1 5 10 15

<210> 193

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR2 of PR000806

<400> 193

Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 194

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> L-FR2 of PR000796, PR001264, PR001265

<400> 194

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr

1 5 10 15

<210> 195

<211> 327

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> human IgG4 constant region

<400> 195

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

1 5 10 15

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

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

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

305 310 315 320

Leu Ser Leu Ser Leu Gly Lys

325

<210> 196

<211> 107

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> human Ig kappa constant region

<400> 196

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 197

<211> 216

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> light chain formula

<220>

<221> X

<222> (1)..(1)

<223> X = D or E

<220>

<221> X

<222> (3)..(3)

<223> X = Q or V

<220>

<221> X

<222> (9)..(9)

<223> X = S or A

<220>

<221> X

<222> (10)..(10)

<223> X = T or S

<220>

<221> X

<222> (13)..(13)

<223> X = A or V

<220>

<221> X

<222> (15)..(15)

<223> X = V or P

<220>

<221> X

<222> (17)..(17)

<223> X = D or E

<220>

<221> X

<222> (19)..(19)

<223> X = V or A

<220>

<221> X

<222> (21)..(21)

<223> X = I or L

<220>

<221> X

<222> (22)..(22)

<223> X = T or S

<220>

<221> X

<222> (29)..(29)

<223> X = I or V

<220>

<221> X

<222> (32)..(32)

<223> X = W, Y or H

<220>

<221> X

<222> (34)..(34)

<223> X = a or N;

<220>

<221> X

<222> (42)..(42)

<223> X = K, N or Q

<220>

<221> X

<222> (45)..(45)

<223> X = K or R

<220>

<221> X

<222> (50)..(50)

<223> X = K, R, V or G

<220>

<221> X

<222> (51)..(51)

<223> X = A or S

<220>

<221> X

<222> (53)..(53)

<223> X = S, T, R or I

<220>

<221> X

<222> (54)..(54)

<223> X = L or R

<220>

<221> X

<222> (55)..(55)

<223> X = E, D, Q or A

<220>

<221> X

<222> (56)..(56)

<223> X21= S or T

<220>

<221> X

<222> (57)..(57)

<223> X = G or D

<220>

<221> X

<222> (58)..(58)

<223> X = V or I

<220>

<221> X

<222> (60)..(60)

<223> X = S or V

<220>

<221> X

<222> (70)..(70)

<223> X = E or D

<220>

<221> X

<222> (72)..(72)

<223> X = T or S

<220>

<221> X

<222> (75)..(75)

<223> X = I or V

<220>

<221> X

<222> (80)..(80)

<223> X = P or S

<220>

<221> X

<222> (81)..(81)

<223> X = D or E

<220>

<221> X

<222> (85)..(85)

<223> X = T or I

<220>

<221> X

<222> (89)..(89)

<223> X = Q or R

<220>

<221> X

<222> (90)..(90)

<223> X = Q, L or H

<220>

<221> X

<222> (91)..(91)

<223> X = Y or-

<220>

<221> X

<222> (92)..(92)

<223> X = N, I, Q or-

<220>

<221> X

<222> (93)..(93)

<223> X = S, T or N

<220>

<221> X

<222> (94)..(94)

<223> X = Y, W, N or Q

<220>

<221> X

<222> (95)..(95)

<223> X = S, T or-

<220>

<221> X

<222> (96)..(96)

<223> X = T or-

<220>

<221> X

<222> (97)..(97)

<223> X = P or-

<220>

<221> X

<222> (98)..(98)

<223> X = Y, R or L

<220>

<221> X

<222> (102)..(102)

<223> X = Q or G

<220>

<221> X

<222> (106)..(106)

<223> X = L or V

<400> 197

Xaa Ile Xaa Met Thr Gln Ser Pro Xaa Xaa Leu Ser Xaa Ser Xaa Gly

1 5 10 15

Xaa Arg Xaa Thr Xaa Xaa Cys Arg Ala Ser Gln Ser Xaa Ser Ser Xaa

20 25 30

Leu Xaa Trp Tyr Gln Gln Lys Pro Gly Xaa Ala Pro Xaa Leu Leu Ile

35 40 45

Tyr Xaa Xaa Ser Xaa Xaa Xaa Xaa Xaa Xaa Pro Xaa Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Xaa Phe Xaa Leu Thr Xaa Ser Ser Leu Gln Xaa

65 70 75 80

Xaa Asp Phe Ala Xaa Tyr Tyr Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

85 90 95

Xaa Xaa Thr Phe Gly Xaa Gly Thr Lys Xaa Glu Ile Lys Arg Thr Val

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

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

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

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

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 198

<211> 442

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<220>

<223> heavy chain general formula

<220>

<221> X

<222> (12)..(12)

<223> X = V or M

<220>

<221> X

<222> (13)..(13)

<223> X = K or Q

<220>

<221> X

<222> (23)..(23)

<223> X = T or S

<220>

<221> X

<222> (27)..(27)

<223> X = G or D

<220>

<221> X

<222> (29)..(29)

<223> X = I, L or V

<220>

<221> X

<222> (30)..(30)

<223> X = S, N, D or Q

<220>

<221> X

<222> (31)..(31)

<223> X = N, S or T

<220>

<221> X

<222> (35)..(35)

<223> X = S or T

<220>

<221> X

<222> (37)..(37)

<223> X = I or F

<220>

<221> X

<222> (40)..(40)

<223> X = P or A

<220>

<221> X

<222> (43)..(43)

<223> X = K or R

<220>

<221> X

<222> (50)..(50)

<223> X = Y or N

<220>

<221> X

<222> (56)..(56)

<223> X = T, N or S

<220>

<221> X

<222> (58)..(58)

<223> X = N or D

<220>

<221> X

<222> (60)..(60)

<223> X = N or Q

<220>

<221> X

<222> (61)..(61)

<223> X = P or S

<220>

<221> X

<222> (64)..(64)

<223> X = K or T

<220>

<221> X

<222> (68)..(68)

<223> X = T or S

<220>

<221> X

<222> (69)..(69)

<223> X = I or M

<220>

<221> X

<222> (71)..(71)

<223> X = V or I

<220>

<221> X

<222> (73)..(73)

<223> X = T or I

<220>

<221> X

<222> (81)..(81)

<223> X = K or N

<220>

<221> X

<222> (83)..(83)

<223> X = S, T or N

<220>

<221> X

<222> (86)..(86)

<223> X = T or I

<220>

<221> X

<222> (87)..(87)

<223> X = A or S

<220>

<221> X

<222> (88)..(88)

<223> X = A or G

<220>

<221> X

<222> (92)..(92)

<223> X = V or M

<220>

<221> X

<222> (96)..(96)

<223> X = A or G

<220>

<221> X

<222> (98)..(98)

<223> X = K or G

<220>

<221> X

<222> (99)..(99)

<223> X = R or K

<220>

<221> X

<222> (100)..(100)

<223> X = G, A, T, W or L

<220>

<221> X

<222> (101)..(101)

<223> X = V, M, G, I or A

<220>

<221> X

<222> (102)..(102)

<223> X = L, Y, H, F or-

<220>

<221> X

<222> (103)..(103)

<223> X = D or S

<220>

<221> X

<222> (105)..(105)

<223> X = W or S

<220>

<221> X

<222> (109)..(109)

<223> X = T or I

<400> 198

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Xaa Val Ser Gly Xaa Ser Xaa Xaa Xaa Tyr

20 25 30

Tyr Trp Xaa Trp Xaa Arg Gln Xaa Pro Gly Xaa Gly Leu Glu Trp Ile

35 40 45

Gly Xaa Ile Tyr Tyr Ser Gly Xaa Thr Xaa Tyr Xaa Xaa Ser Leu Xaa

50 55 60

Ser Arg Val Xaa Xaa Ser Xaa Asp Xaa Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Xaa Leu Xaa Ser Val Xaa Xaa Xaa Asp Thr Ala Xaa Tyr Tyr Cys Xaa

85 90 95

Arg Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Gly Gln Gly Xaa Leu Val Thr

100 105 110

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

115 120 125

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

130 135 140

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

145 150 155 160

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly

165 170 175

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly

180 185 190

Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

195 200 205

Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys

210 215 220

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

225 230 235 240

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

245 250 255

Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp

260 265 270

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

275 280 285

Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

290 295 300

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

305 310 315 320

Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

325 330 335

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

340 345 350

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

355 360 365

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

370 375 380

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

385 390 395 400

Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn

405 410 415

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

420 425 430

Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

Claims

1. An isolated CD47 antibody or antigen-binding fragment thereof, comprising an antibody heavy chain variable region VH comprising HCDR1, HCDR2 and HCDR3 and an antibody light chain variable region VL comprising LCDR1, LCDR2 and LCDR3, wherein the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are selected from any one of the group consisting of:

(1) HCDR 1: 36, HCDR 2: 138, HCDR 3: 58, LCDR 1: 115, LCDR 2: 31 and LCDR 3: 70 in SEQ ID NO;

(2) HCDR 1: 32, HCDR 2: 138, HCDR 3: 58, LCDR 1: 115, LCDR 2: 31 and LCDR 3: 70 in SEQ ID NO; and

(3) HCDR 1: 34, HCDR 2: 138, HCDR 3: 58, LCDR 1: 115, LCDR 2: 31 and LCDR 3: 70 in SEQ ID NO.

2. The isolated CD47 antibody or antigen binding fragment thereof of claim 1, wherein the antigen binding fragment comprises a Fab, Fab ', Fv fragment, F (ab')₂scFv, di-scFv and/or dAb.

3. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody is selected from the group consisting of: chimeric antibodies, humanized antibodies, and fully human antibodies.

4. The isolated CD47 antibody or antigen-binding fragment thereof of claim 1, wherein the VH comprises the framework regions H-FR1, H-FR2, H-FR3, and H-FR 4.

5. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 4, wherein the C-terminus of H-FR1 is linked directly or indirectly to the N-terminus of the HCDR1 and the H-FR1 comprises the amino acid sequence set forth in SEQ ID NO 145.

6. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 5, wherein the H-FR1 comprises the amino acid sequence set forth in SEQ ID NO: 170.

7. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 4, wherein the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises the amino acid sequence set forth in SEQ ID NO 146.

8. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 7, wherein the H-FR2 comprises the amino acid sequence set forth in SEQ ID No. 190.

9. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 4, wherein the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises the amino acid sequence set forth in SEQ ID NO: 147.

10. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 9, wherein the H-FR3 comprises the amino acid sequence set forth in SEQ ID No. 176.

11. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 4, wherein the N-terminus of H-FR4 is linked directly or indirectly to the C-terminus of the HCDR3 and the H-FR4 comprises the amino acid sequence set forth in SEQ ID NO 148.

12. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 11, wherein the H-FR4 comprises the amino acid sequence set forth in SEQ ID NO: 186.

13. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 1, wherein the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs 111, 107, and 109.

14. The isolated CD47 antibody or antigen-binding fragment thereof of claim 1, comprising an antibody heavy chain constant region, and the antibody heavy chain constant region comprises a human IgG constant region.

15. The isolated CD47 antibody or antigen-binding fragment thereof of claim 1, comprising an antibody heavy chain constant region, and the antibody heavy chain constant region comprises a human IgG4 constant region.

16. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 15, wherein the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID No. 195.

17. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 1, comprising an antibody heavy chain HC, and the HC comprises an amino acid sequence set forth in any one of SEQ ID NOs 112, 108, and 110.

18. The isolated CD47 antibody or antigen-binding fragment thereof of claim 1, wherein the VL comprises the framework regions L-FR1, L-FR2, L-FR3, and L-FR 4.

19. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 18, wherein the C-terminus of L-FR1 is linked directly or indirectly to the N-terminus of the LCDR1 and the L-FR1 comprises the amino acid sequence set forth in SEQ ID No. 149.

20. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 19, wherein the L-FR1 comprises the amino acid sequence set forth in SEQ ID No. 158.

21. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 18, wherein the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises the amino acid sequence set forth in SEQ ID No. 150.

22. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 21, wherein the L-FR2 comprises the amino acid sequence set forth in SEQ ID No. 194.

23. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 18, wherein the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises the amino acid sequence set forth in SEQ ID No. 151.

24. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 23, wherein the L-FR3 comprises the amino acid sequence set forth in SEQ ID No. 155.

25. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 18, wherein the N-terminus of L-FR4 is linked directly or indirectly to the C-terminus of the LCDR3 and the L-FR4 comprises the amino acid sequence set forth in SEQ ID No. 152.

26. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 25, wherein the L-FR4 comprises the amino acid sequence set forth in SEQ ID No. 159.

27. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 1, wherein the VL comprises the amino acid sequence set forth in SEQ ID No. 29.

28. The isolated CD47 antibody or antigen-binding fragment thereof of claim 1, comprising an antibody light chain constant region, and the antibody light chain constant region comprises a human Ig kappa constant region.

29. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 28, wherein the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID No. 196.

30. The isolated CD47 antibody or antigen-binding fragment thereof according to claim 1, comprising an antibody light chain LC, and the LC comprises the amino acid sequence set forth in SEQ ID No. 30.

31. One or more isolated nucleic acid molecules encoding the isolated CD47 antibody or antigen-binding fragment thereof of any one of claims 1-30.

32. A vector comprising the nucleic acid molecule of claim 31.

33. A cell comprising the nucleic acid molecule of claim 31 or the vector of claim 32.

34. A method of making the isolated CD47 antibody or antigen-binding fragment thereof of any one of claims 1-30, the method comprising culturing the cell of claim 33 under conditions such that the isolated CD47 antibody or antigen-binding fragment thereof of any one of claims 1-30 is expressed.

35. A pharmaceutical composition comprising the isolated CD47 antibody or antigen-binding fragment thereof of any one of claims 1-30, the nucleic acid molecule of claim 31, the vector of claim 32, and/or the cell of claim 33, and optionally a pharmaceutically acceptable carrier.