CN109384846B

CN109384846B - Antibody capable of binding TIGIT (tungsten inert gas) or antigen-binding fragment thereof and application

Info

Publication number: CN109384846B
Application number: CN201811116733.6A
Authority: CN
Inventors: 田志刚; 曹国帅; 王保如; 孙汭; 肖卫华; 魏海明; 王立
Original assignee: Immunopharmaceutic Institute Of Hefei Ruida Co ltd
Current assignee: Immunopharmaceutic Institute Of Hefei Ruida Co ltd
Priority date: 2018-09-25
Filing date: 2018-09-25
Publication date: 2020-03-03
Anticipated expiration: 2038-09-25
Also published as: CN109384846A

Abstract

An antibody or antigen-binding fragment thereof capable of binding to TIGIT and uses thereof. The invention relates to the field of antibodies, and discloses an antibody capable of binding TIGIT and application thereof. The antibody includes a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3, as well as a light chain CDR1, a light chain CDR2, and a light chain CDR 3. The antibodies of the invention have the following properties: ability to block the interaction of TIGIT and CD155 with high affinity; capable of binding with high specificity to TIGIT, but not to other CD28 family members (e.g., CD28, PD-1, and CTLA 4); activate immune cells, thereby killing tumor cells.

Description

Antibody capable of binding TIGIT (tungsten inert gas) or antigen-binding fragment thereof and application

Technical Field

The invention relates to the field of antibodies, in particular to an antibody capable of binding TIGIT or an antigen binding fragment thereof and application.

Background

Tumor immunotherapy is a long-standing hotspot in the field of tumor therapy. Strategies that use antibodies to block immunosuppressive receptors (including PD-1, CTLA4, etc.) have met with great success in clinical tumor therapy, but a significant proportion of patients do not respond to PD-1, CTLA4, and combinations of both, probably due to the expression of multiple immunosuppressive receptors on the surface of T cells, NK cells. The inhibitory receptors such as PD-1, CTLA4 and TIGIT may act synergistically to inhibit the activation of immune cells including T cells and NK cells, and therefore, the development of therapeutic antibodies against the immunosuppressive receptors such as TIGIT is of great significance.

Tigit (T cell Ig and ITIM domain) molecules are inhibitory receptors expressed on the surface of T cells, NK cells found in 2009, and mainly comprise an extracellular IgV-like domain, a transmembrane region, and an intracellular segment containing an ITIM motif (immunoreceptor tyrosine-based inhibitory motif). TIGIT binds ligand PVR (CD 155) with high affinity and transmits inhibitory signals through intracellular ITIM motifs, thereby inhibiting the function of T cells, NK cells.

Studies have shown that TIGIT expression is associated with tumor development. TIGIT molecules are highly expressed on the surface of tumor infiltration T cells of lung cancer patients, and the expression of the TIGIT molecules is in a positive correlation with the expression of PD-1. The TIGIT expression on the surface of tumor infiltrating NK cells of colon cancer patients is obviously higher than that of peripheral blood NK cells. In various mouse tumor models, blocking TIGIT by using the antibody can inhibit tumor growth and metastasis, and blocking TIGIT and PD-1 can effectively treat tumors. Therefore, development of anti-TIGIT antibodies is particularly necessary.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide an antibody capable of mediating various anti-tumor mechanisms and application thereof.

The antibody provided by the invention can provide the patient self immune response to the tumor by blocking the TIGIT/CD155 combination. In a humanized mouse tumor model, the antibody of the present invention has the effect of inhibiting tumor growth, which indicates that the antibody of the present invention has positive therapeutic effects on tumors.

Accordingly, to achieve the above objects, in a first aspect, the present invention provides an antibody or antigen binding fragment thereof capable of binding to TIGIT, characterized in that the antibody comprises heavy chain CDR1, heavy chain CDR2 and heavy chain CDR3, and light chain CDR1, light chain CDR2 and light chain CDR 3;

wherein the amino acid sequence of the heavy chain CDR1 has at least 80% homology with the sequence shown by SEQ ID NO 2, SEQ ID NO 10, SEQ ID NO 20, SEQ ID NO 26, SEQ ID NO 32, SEQ ID NO 39, SEQ ID NO 50, SEQ ID NO 57, SEQ ID NO 59 or SEQ ID NO 79,

the amino acid sequence of the heavy chain CDR2 has at least 80% homology with the sequence shown by SEQ ID NO. 3, SEQ ID NO. 11, SEQ ID NO. 21, SEQ ID NO. 27, SEQ ID NO. 37, SEQ ID NO. 40, SEQ ID NO. 51, SEQ ID NO. 60, SEQ ID NO. 73 or SEQ ID NO. 80,

the amino acid sequence of the heavy chain CDR3 has at least 80% homology with the sequences shown by SEQ ID NO. 4, SEQ ID NO. 12, SEQ ID NO. 16, SEQ ID NO. 22, SEQ ID NO. 28, SEQ ID NO. 33, SEQ ID NO. 41, SEQ ID NO. 52, SEQ ID NO. 62, SEQ ID NO. 66, SEQ ID NO. 77, SEQ ID NO. 81 or SEQ ID NO. 87,

the amino acid sequence of the light chain CDR1 has at least 80% homology with the sequence shown by SEQ ID NO 6, SEQ ID NO 14, SEQ ID NO 18, SEQ ID NO 30, SEQ ID NO 35, SEQ ID NO 43, SEQ ID NO 47, SEQ ID NO 54, SEQ ID NO 68, SEQ ID NO 83 or SEQ ID NO 85,

the amino acid sequence of the light chain CDR2 has at least 80% homology with the sequence shown in SEQ ID NO. 7, SEQ ID NO. 44 or SEQ ID NO. 69,

the amino acid sequence of the light chain CDR3 has at least 80% homology with the sequence shown in SEQ ID NO 8, SEQ ID NO 24, SEQ ID NO 48, SEQ ID NO 55, SEQ ID NO 64, SEQ ID NO 70 or SEQ ID NO 75.

In a second aspect, the present invention provides an immunoconjugate comprising a therapeutic agent and the antibody or antigen-binding fragment thereof of the first aspect coupled to the therapeutic agent.

In a third aspect, the present invention provides a composition comprising an antibody or antigen-binding fragment thereof according to the first aspect, and/or an immunoconjugate as described above, and a pharmaceutically acceptable carrier.

In a fourth aspect, the invention provides a kit for detecting TIGIT in a sample, the kit comprising an antibody or antigen-binding fragment thereof of the first aspect.

In a fifth aspect, the invention provides use of the antibody or antigen-binding fragment thereof of the first aspect in the preparation of a reagent for detecting TIGIT in a sample.

In a sixth aspect, the invention provides use of the antibody or antigen-binding fragment thereof of the first aspect in the manufacture of a medicament for the prevention and/or treatment of TIGIT-mediated diseases.

In a seventh aspect, the invention provides a nucleic acid encoding the antibody or antigen-binding fragment thereof of the first aspect.

In an eighth aspect, the present invention provides a recombinant vector or transformant containing the nucleic acid according to the seventh aspect.

According to the technical scheme, the invention provides the anti-TIGIT antibody or the antibody fragment capable of being combined with TIGIT. The antibodies of the invention have at least one of the following properties: ability to block the interaction of TIGIT and CD155 with high affinity; capable of binding with high specificity to TIGIT, but not to other CD28 family members (e.g., CD28, PD-1, and CTLA 4); activate immune cells, thereby killing tumor cells.

Additional features and advantages of the invention will be set forth in the detailed description which follows.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:

fig. 1 is a graph of ELISA results for TIGIT antibodies according to embodiments of the invention;

FIG. 2 is a graph of the results of TIGIT antibody binding to 293T-TIGIT cells according to an embodiment of the present invention;

FIG. 3 is a graph of the results of TIGIT antibody blocking ligand CD155 binding according to embodiments of the present invention;

FIG. 4 is a graph showing the results of binding of TIGIT antibody to human natural killer cells according to an embodiment of the present invention;

fig. 5 is a graph showing the results of TIGIT antibodies according to embodiments of the invention not binding to mouse TIGIT protein;

FIG. 6 is a graph of the results of a TIGIT antibody not binding to other proteins of the same family according to an embodiment of the present invention;

FIG. 7 is a graph of the results of TIGIT antibody competition according to an embodiment of the present invention;

FIG. 8 is a graph of the anti-cancer function of human peripheral blood mononuclear cells promoted by TIGIT antibodies according to an embodiment of the present invention;

FIG. 9 is a graph of TIGIT antibody inhibiting the growth of colon cancer SW620 cells in accordance with an embodiment of the present invention;

FIG. 10 is a graph of the results of binding of a humanized TIGIT antibody to 293T-TIGIT cells according to an embodiment of the present invention;

fig. 11 is a graph of the results of humanized TIGIT antibodies blocking ligand CD155 binding according to embodiments of the invention.

Detailed Description

The following describes in detail specific embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the present invention, are given by way of illustration and explanation only, not limitation.

The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.

In order that the invention may be more readily understood, certain technical and scientific terms are specifically defined below. Unless clearly defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The abbreviations for amino acid residues are standard 3-letter and/or 1-letter codes used in the art to refer to one of the 20 commonly used L-amino acids.

In bookIn the present invention, where the term antigen-binding fragment is used without a contrary indication, an "antibody fragment", which generally refers to an antigen-binding antibody fragment, may include a portion of an intact antibody, typically an antigen-binding or variable region, examples of which include Fab, Fab ', F (ab')₂Fv or scFv, diabodies, linear antibodies, single-chain antibody molecules, and the like.

The term "complementarity determining region" or "CDR sequence" refers to the amino acid sequence of an antibody that is responsible for antigen binding, e.g., generally including: amino acid residues in the light chain variable region near 23-34(L1), 50-56(L2) and 89-97(L3), and in the heavy chain variable region near 31-35B (H1), 50-65(H2) and 95-102(H3) (Kabat et al, Sequences of proteins of Immunological Interest, 5^thPublic Health Service, national institutes of Health, Bethesda, MD. (1991)); and/or from "hypervariable loops" (e.g., amino acid residues in the light chain variable region near 26-32(LI), 50-52(L2), and 91-96(L3), and in the heavy chain variable region near 26-32(H1), 53-55(H2), and 96-101(H3) (Chothia and Lesk J.mol.biol.196: 901-917 (1987)).

One skilled in the art can substitute, add and/or delete one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 or more) amino acids to the sequences of the invention to obtain variants of the sequences of the antibodies or functional fragments thereof, without substantially affecting the activity of the antibodies (retaining at least 95% of the activity). All of which are considered to be included within the scope of the present invention. Such as the substitution of amino acids with similar properties in the variable region. The sequence of a variant of the invention may have at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity (or homology) to a reference sequence. The sequence identity described in the present invention can be measured using sequence analysis software. For example the computer program BLAST, in particular BLASTP or TBLASTN, using default parameters. The amino acid sequences mentioned in the present invention are shown from N-terminus to C-terminus.

As previously noted, the antibodies of the invention can be full-length (e.g., IgG1 or IgG4 antibodies) or can comprise only antigen-binding portions (e.g.E.g., Fab, F (ab')₂Or scFv fragments) or may be modified to affect function. The invention includes anti-TIGIT antibodies with modified glycosylation patterns. In some applications, it may be useful to modify to remove undesired glycosylation sites, or antibodies that do not have a fucose moiety on the oligosaccharide chain, for example, to enhance antibody-dependent cellular cytotoxicity (ADCC) function. In other applications, galactosylation modifications can be made to alter Complement Dependent Cytotoxicity (CDC).

The term "functional fragment" as used herein refers in particular to antibody fragments such as Fv, scFv (sc for single chain), Fab, F (ab')₂Fab ', scFv-Fc fragment or diabody (diabody), or any fragment which should be able to increase half-life by chemical modification, e.g. addition of a poly (alkylene) glycol, such as polyethylene glycol ("PEGylation", PEGylation ") (known as Fv-PEG, scFv-PEG, Fab-PEG, F (ab')₂-pegylated fragments of PEG or Fab' -PEG) ("PEG" is polyethylene glycol), said fragments having TIGIT binding activity. Preferably, the functional fragment will consist of or comprise a partial sequence of the heavy chain variable region or the light chain variable region of the antibody from which it is derived sufficient to retain the same binding specificity and sufficient affinity as the antibody from which it is derived, preferably 1/100 at least equal to the affinity of the antibody from which it is derived, and more preferably 1/10 in a more preferred manner. Such functional fragments will comprise a minimum of 5 amino acids, preferably 10, 15, 25, 50 and 100 consecutive amino acids of the antibody sequence from which they are derived.

As previously described, the present invention provides an antibody or antigen binding fragment thereof capable of binding TIGIT, wherein the antibody comprises heavy chain CDR1, heavy chain CDR2 and heavy chain CDR3, and light chain CDR1, light chain CDR2 and light chain CDR 3. Preferably, the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2 and light chain CDR3 are at least 80% homologous to the sequences shown in the columns of Table 1, simultaneously.

Preferably, each CDR sequence has 100% identity (homology) with a reference sequence, i.e., the amino acid sequence of the heavy chain CDR1 is as shown in SEQ ID NO 2, SEQ ID NO 10, SEQ ID NO 20, SEQ ID NO 26, SEQ ID NO 32, SEQ ID NO 39, SEQ ID NO 50, SEQ ID NO 57, SEQ ID NO 59 or SEQ ID NO 79,

and/or the amino acid sequence of the heavy chain CDR2 is shown in SEQ ID NO. 3, SEQ ID NO. 11, SEQ ID NO. 21, SEQ ID NO. 27, SEQ ID NO. 37, SEQ ID NO. 40, SEQ ID NO. 51, SEQ ID NO. 60, SEQ ID NO. 73 or SEQ ID NO. 80,

and/or the amino acid sequence of the heavy chain CDR3 is shown in SEQ ID NO. 4, SEQ ID NO. 12, SEQ ID NO. 16, SEQ ID NO. 22, SEQ ID NO. 28, SEQ ID NO. 33, SEQ ID NO. 41, SEQ ID NO. 52, SEQ ID NO. 62, SEQ ID NO. 66, SEQ ID NO. 77, SEQ ID NO. 81 or SEQ ID NO. 87,

and/or the amino acid sequence of the light chain CDR1 is shown in SEQ ID NO 6, SEQ ID NO 14, SEQ ID NO 18, SEQ ID NO 30, SEQ ID NO 35, SEQ ID NO 43, SEQ ID NO 47, SEQ ID NO 54, SEQ ID NO 68, SEQ ID NO 83 or SEQ ID NO 85,

and/or the amino acid sequence of the light chain CDR2 is shown in SEQ ID NO 7, SEQ ID NO 44 or SEQ ID NO 69,

and/or the amino acid sequence of the light chain CDR3 is shown in SEQ ID NO 8, SEQ ID NO 24, SEQ ID NO 48, SEQ ID NO 55, SEQ ID NO 64, SEQ ID NO 70 or SEQ ID NO 75.

According to a more preferred embodiment of the invention, the CDR sequences are as shown in the columns of Table 1, i.e. the amino acid sequence of the heavy chain CDR1 is shown in SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown in SEQ ID NO. 3 and the amino acid sequence of the heavy chain CDR3 is shown in sequence Listing SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 12;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 16;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 20, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 21 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 22;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 26, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 28;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 32, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 33;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 37 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 39, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 40 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 41;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 50, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 51, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 52;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 57, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 59, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 60 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 62;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 66;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 26, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 73, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 77;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 79, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 80, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 81;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 87;

and/or the presence of a gas in the gas,

the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 14, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 18, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 24;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 35, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 43, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 44 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO:47, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO:7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO: 48;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 54, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 55;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 64;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 68, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 69 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 70;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 75;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 83, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 44, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 85, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8.

According to a further preferred embodiment of the present invention, the CDR sequences each simultaneously satisfy the amino acid sequences shown in the columns of Table 1, i.e. the amino acid sequence of the heavy chain CDR1 is shown in SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown in SEQ ID NO. 3 and the amino acid sequence of the heavy chain CDR3 is shown in sequence Listing SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 12; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 14, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 16; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 18, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 20, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 21 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 22; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 24;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 26, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 28; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 32, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 33; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 35, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 37 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 39, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 40 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 41; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 43, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 44 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO:47, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO:7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO: 48;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 50, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 51, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 52; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 54, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 55;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 57, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 59, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 60 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 62; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 64;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 66; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 68, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 69 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 70;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 26, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 73, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 75;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 77; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 64;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 3 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 2, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 30, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 79, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 80, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 81; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 83, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 44 and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 26, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 27 and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 4; the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 85, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8;

or the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 10, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 11, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 87; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 35, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 8.

The antibody comprises a heavy chain variable region and a light chain variable region, and the amino acid sequences of the heavy chain variable region or the light chain variable region of the antibody of the present invention may have at least 80% homology with the sequences shown in table 1, respectively. That is, according to a particularly preferred embodiment of the present invention, the heavy chain variable region of the antibody has the amino acid sequence shown in SEQ ID NO 1, SEQ ID NO 9, SEQ ID NO 15, SEQ ID NO 19, SEQ ID NO 25, SEQ ID NO 31, SEQ ID NO 36, SEQ ID NO 38, SEQ ID NO 45, SEQ ID NO 49, SEQ ID NO 56, SEQ ID NO 58, SEQ ID NO 61, SEQ ID NO 65, SEQ ID NO 71, SEQ ID NO 72, SEQ ID NO 76, SEQ ID NO 78 or SEQ ID NO 86; and/or the amino acid sequence of the light chain variable region of the antibody is shown as SEQ ID NO 5, SEQ ID NO 13, SEQ ID NO 17, SEQ ID NO 23, SEQ ID NO 29, SEQ ID NO 34, SEQ ID NO 42, SEQ ID NO 46, SEQ ID NO 53, SEQ ID NO 63, SEQ ID NO 67, SEQ ID NO 74, SEQ ID NO 82 or SEQ ID NO 84.

Preferably, the amino acid sequences of the heavy chain variable region and the light chain variable region of the antibody of the present invention have at least 80% homology to the sequences shown in table 1 at the same time. That is, according to a particularly preferred embodiment of the present invention, the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 1 and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 5;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 9 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 13;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 15 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 17;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 19 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 23;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 25 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 31 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 34;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 36 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO 38 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO 42;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 45 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 46;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 49 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 53;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 56 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 58 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 61 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 63;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 65 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 67;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 71 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 72 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 74;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO:76 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 63;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 1 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 45 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 29;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO:78 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 82;

or, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO:71 and the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 84;

or the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 86, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 34.

TABLE 1 amino acid sequence numbering of antibodies

Note: the numbers in Table 1 are the numbers of "SEQ ID NO" in the sequence Listing.

According to the invention, the antibody is capable of binding to TIGIT (antigen), in particular the amino acid sequence shown in SEQ ID NO: 121.

In a preferred embodiment of the invention, the antibody may also be humanized, i.e. the antibody is a chimeric or humanized antibody, in order to further improve the bioavailability of the antibody. The term "chimeric antibody" refers to a recombinant antibody obtained by replacing the amino acid sequence of the constant region of a monoclonal antibody from one species (e.g., mouse) with the constant region of an antibody from another species (e.g., human) using recombinant DNA techniques (the heavy chain constant region is shown in SEQ ID NO:124 and the light chain constant region is shown in SEQ ID NO: 125). The term "humanized antibody" refers to a recombinant antibody obtained by replacing all of the amino acid sequences of the non-CDR (Fv Framework Region (FR)) of the constant region and variable region of a monoclonal antibody from one species (e.g., mouse) with the amino acid sequences of the non-CDR of the constant region and variable region of an antibody from another species (e.g., human) using recombinant DNA techniques. That is, the constant region of an antibody is called a chimeric antibody when it is humanized, and the non-CDR amino acid sequences of the constant region and the variable region are all humanized and called a humanized antibody. The humanization method can be performed by referring to conventional antibody engineering techniques, and will not be described herein.

For example, the amino acid sequence of the chimeric antibody provided by the invention can be as follows SEQ ID NO:126 (heavy chain) and SEQ ID NO:127 (light chain).

The heavy chain variable region sequence of the humanized antibody provided by the invention is shown as SEQ ID NO. 122, and the light chain variable region sequence is shown as SEQ ID NO. 123.

The immunoconjugates provided herein comprise a therapeutic agent and an antibody or antigen-binding fragment thereof as described above coupled to the therapeutic agent. The manner in which the antibody or antigen-binding fragment thereof is conjugated to the therapeutic agent may be conventional.

The present invention provides compositions comprising an antibody or antigen-binding fragment thereof as described above, and/or an immunoconjugate as described above, and a pharmaceutically acceptable carrier. In certain embodiments, the compositions include temporally and/or spatially separated combinations, so long as they can act together to achieve the objectives of the present invention. For example, the ingredients contained in the composition may be administered to the subject in bulk, or separately. When the ingredients contained in the composition are administered separately to a subject, the individual ingredients may be administered to the subject simultaneously or sequentially.

The term "pharmaceutically acceptable" indicates that the composition can be administered to a subject without producing adverse physiological reactions that interfere with administration of the composition. For example, "pharmaceutically acceptable carrier" refers to a carrier useful in preparing generally safe, non-toxic, desirable pharmaceutical compositions, preferably, examples of such carriers or diluents include, but are not limited to: water, saline, ringer's solution, dextrose, mannitol, dextrose, lactose, starch, magnesium stearate, cellulose, magnesium carbonate, 0.3% glycerol, hyaluronic acid, ethanol, polyalkylene glycols such as polypropylene glycol, triglycerides, 5% human serum albumin, liposomes and nonaqueous vehicles, such as fixed oils, may also be used.

The compositions of the invention may also be administered in combination with each other, or with one or more other therapeutic compounds, for example, in combination with a chemotherapeutic agent. Thus, the composition may also contain a chemotherapeutic agent. The antibodies or antigen-binding fragments thereof, or immunoconjugates of the invention can also be combined with a second therapeutic agent, exemplary agents of which include, but are not limited to, other agents that inhibit TIGIT activity (including other antibodies or antigen-binding fragments thereof, peptide inhibitors, small molecule antagonists, etc.) and/or agents that interfere with TIGIT upstream or downstream signaling.

Typically, the antibody or antigen-binding fragment thereof is administered in an effective amount, i.e., an amount sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g., an amount that causes prevention or alleviation of symptoms associated with the disease being treated, e.g., TIGIT-associated disease. The effective amount of the composition administered to a subject will depend on the type and severity of the disease, as well as on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs; it will also depend on the severity and type of the disease and the skilled person will be able to determine the appropriate dosage depending on such factors and the like.

The kit for detecting TIGIT in a sample provided by the invention contains the antibody or the antigen-binding fragment thereof. The sample may be tissue of a patient suffering from TIGIT-mediated disease (particularly a cancer patient, more preferably a patient suffering from at least one of lung cancer, liver cancer, ovarian cancer, cervical cancer, skin cancer, bladder cancer, colon cancer, breast cancer, glioma, renal cancer, gastric cancer, esophageal cancer, oral squamous cell carcinoma, and head and neck cancer). The kit may also include reagents conventionally used for detection of TIGIT, such as coating solutions and the like.

The invention also provides the use of an antibody or antigen-binding fragment thereof as described above in the preparation of a reagent for the detection of TIGIT in a sample. As previously mentioned, the sample may be tissue of a patient suffering from TIGIT-mediated disease and will not be described herein. The antibody or the antigen-binding fragment thereof of the present invention has good affinity for TIGIT, and can efficiently detect TIGIT in a sample.

The invention also provides application of the antibody or the antigen-binding fragment thereof in preparing a medicament for preventing and/or treating TIGIT-mediated diseases. Preferably, the TIGIT-mediated disease is cancer or an infectious disease. More preferably, the cancer is a CD155 expressing cancer. Further preferably, the cancer is at least one of lung cancer, liver cancer, ovarian cancer, cervical cancer, skin cancer, bladder cancer, colon cancer, breast cancer, glioma, kidney cancer, stomach cancer, esophageal cancer, oral squamous cell carcinoma and head and neck cancer. Further preferably, the infectious disease includes, but is not limited to, HIV virus infection and/or hepatitis b virus infection.

In addition, the present invention relates to a method of inhibiting (in vitro) cancer cells, the method comprising: contacting a cancer cell (as described above) with an effective amount of at least one of an antibody or antigen-binding fragment thereof, an immunoconjugate, and a composition of the invention. The term "inhibiting" as used herein includes inhibiting the proliferation, growth and/or metastasis of cancer cells.

The present invention also relates to a method of preventing and/or treating TIGIT mediated diseases (as described above) comprising: administering to the patient an effective amount of at least one of the antibody or antigen-binding fragment thereof, immunoconjugate and composition of the invention. The administration mode can be oral administration, nasal administration, intradermal administration, subcutaneous administration, intramuscular administration or intravenous administration or intraperitoneal administration.

As used herein, reference to a "patient" or "subject" refers generally to a mammal, such as a primate and/or rodent, and particularly a human or mouse.

One skilled in the art can clone a DNA molecule encoding the antibody or antigen-binding fragment thereof of the present invention into a vector (particularly an expression vector) and further transform the host cell, and the antibody or antigen-binding fragment thereof can be obtained by inducible expression. Accordingly, the present invention also provides (isolated) nucleic acids encoding the above-described antibodies or antigen-binding fragments thereof, as well as recombinant vectors and transformants containing the nucleic acids. The nucleic acid is preferably an expression cassette obtained by genetic engineering means.

The recombinant vector may be a cloning vector or an expression vector, and may be obtained by operably linking the nucleic acid with a commercially available vector (e.g., a plasmid or a viral vector), and commonly used plasmids include pSeTag2, PEE14, pMH3, and the like.

The expression vector of the present invention may contain a DNA sequence encoding the heavy chain variable region, the light chain variable region and/or the constant region of the antibody. However, it is also possible to construct two expression vectors, one containing the heavy chain variable region and constant region and the other containing the light chain variable region and constant region, separately, and to transfect mammalian cells together. In a preferred embodiment, the expression vector further comprises a promoter and a DNA sequence encoding a secretion signal peptide, and at least one drug-resistant gene for screening.

The host cell of the invention may be a prokaryotic host cell, a eukaryotic host cell or a bacteriophage. The prokaryotic host cell can be escherichia coli, bacillus subtilis, streptomyces or proteus mirabilis and the like. The eukaryotic host cell can be fungi such as Pichia pastoris, saccharomyces cerevisiae, schizosaccharomyces and trichoderma, insect cells such as meadow armyworm, plant cells such as tobacco, and mammalian cells such as BHK cells, CHO cells, COS cells and myeloma cells. In some embodiments, the host cell of the invention is preferably a mammalian cell, more preferably a BHK cell, a CHO cell, an NSO cell or a COS cell.

Nucleic acids encoding the heavy and/or light chains of the antibodies of the invention are within the scope of the invention, and depending on the amino acid sequence of the heavy and/or light chain, the corresponding nucleic acid sequences are readily available to those skilled in the art, as shown in table 2.

TABLE 2 Gene sequence numbering of antibodies

Note: the numbers in Table 2 are the numbers of "SEQ ID NO" in the sequence Listing.

The present invention will be described in detail below by way of examples. In the examples and test examples, the experimental methods not specified in specific conditions were carried out under the usual conditions.

Example 1: preparation of antibodies

A murine monoclonal antibody against human TIGIT was generated, and Balb/c mice (9 weeks old, purchased from Shanghai Leisi, approximately 20g in weight) were immunized with purified recombinant TIGIT extracellular region Fc fusion protein (TIGIT-Fc) (recombinant TIGIT extracellular region Fc fusion protein, amino acid sequence shown in SEQ ID NO:121) as an antigen.

The immunized mice were immunized 3 times with purified antigen and complete Freund's adjuvant and the immune response was detected after bleeding through the tail vein. And screening serum by ELISA and flow cytometry to obtain the mouse with the anti-human TIGIT immunoglobulin. And splenocytes from mice with the highest anti-TIGIT immunoglobulins were fused with murine myeloma SP2/0 cells (ATCC accession number CRL-1581). The fused hybridoma cells are subjected to antibody screening to obtain the murine monoclonal antibodies 1H6, 2B11, 3A10, 4A5, 4A9, 4H5, 6A2, 6B7, 7F4, 8E1, 8G3, 9F4, 9G6, 10C1, 10F10, 11G4, 12B7, 12C8, 15E9, 16C11, 16D6 and 16E 10.

Culturing the total number of candidate hybridoma cells to 10⁶Cells were harvested by centrifugation at 800rpm for 10 minutes and total RNA was extracted using Trizol kit (Invitrogen); the total RNA is used as a template to synthesize a cDNA library (Invitrogen) through reverse transcription, and the cDNA is used as a template to amplify the variable region nucleic acid sequence corresponding to the hybridoma cells through PCR. The primer sequences used in the PCR amplification reaction are complementary to the first framework region of the antibody variable region or the signal peptide region and the constant region (Larrick, J.W., et al., (1990) Scand.J.Immunol., 32, 121-. Mu.l of cDNA 2. mu.l, 5. mu.l of 10 XPCR buffer, 2. mu.l (5. mu. mol) of upstream and downstream primers, 2. mu.l of dNTP, 1. mu.l of Taq enzyme (Takara, Ex Taq), H, were added to a 50. mu.l reaction system₂O38 μ l; pre-denaturation at 95 ℃ for 5min, and performing PCR amplification in a temperature cycle. The reaction conditions are as follows: denaturation at 94 ℃ for 30S, annealing at 58 ℃ for 45S, and extension at 72 ℃ for 50S for 32 cycles, followed by extension at 72 ℃ for 7 min. Sequencing the amplified product to obtain mouse monoclonal antibodyThe heavy and light chain variable region sequences (including amino acid sequences and nucleic acid sequences) are shown in tables 1 and 2.

Example 2: antibody binding Capacity screening

TIGIT antibody ELISA binding experiment

ELISA experiments were used to detect the binding properties of TIGIT antibodies. And (3) coating the Fc fusion protein of the extracellular region of TIGIT (TIGIT-Fc) into a 96-well plate, and judging the binding property of the antibody and the TIGIT by using the intensity of a signal after the antibody is added.

TIGIT-Fc fusion protein (amino acid sequence shown in SEQ ID NO:121) was diluted to 1. mu.g/ml with PBS buffer, added to a 96-well plate in a volume of 100. mu.l/well, and left overnight at 4 ℃. The 96-well plate was aspirated off PBS buffer, and after washing the plate 6 times with PBST (pH7.2PBS containing 0.1% Tween 20) buffer, 200. mu.l/well PBS/10% BSA was added and incubated at 37 ℃ for 2h for blocking. The blocking solution was removed, and after washing the plate 6 times with PBST, the TIGIT antibody to be tested diluted to an appropriate concentration with PBST/0.05% BSA at 100. mu.l/well was added, and incubated at 37 ℃ for 1 h. The reaction was removed and after washing the plate 6 times with PBST, HRP (horseradish peroxidase) -labeled anti-mouse antibody secondary antibody was diluted with PBST/0.05% BSA at 100. mu.l/well and incubated at 37 ℃ for 1 h. After washing the plate 6 times with PBST, 80. mu.l/well of TMB (tetramethylbenzidine) was added, the plate was incubated at room temperature for 3min, and the reaction was stopped by adding 80. mu.l/well of 4M sulfuric acid. The absorbance was read at 450mm using a microplate reader. The results (fig. 1) indicate that the antibodies of the invention are capable of binding TIGIT.

TIGIT antibody flow cytometry binding experiment

Flow cytometry experiments were used to detect the binding properties of TIGIT antibodies, where TIGIT protein (HEK293T-TIGIT) was overexpressed in HEK293T cells (ATCC accession number CRL-3249), and the intensity of the signal after antibody addition was used to determine the binding properties of the antibodies to TIGIT.

HEK293T cells at 5X 10⁵Cells/well were plated in six-well plates and incubated overnight in DMEM medium without double antibody. Media was discarded before transfection and 1ml of fresh DMEM media without double antibody was added. The coding sequence (shown as SEQ ID NO: 128) of TIGIT protein (SEQ ID NO:121), pMD2G and psPAX2 vector are inserted between EcoRI and BamHI of the restriction site of pLVX-TIGIT-IRES-ZsGreen (pLVX-IRES-ZsGreen1 vector), and the vector thereof(total 3. mu.g) 200. mu.l serum-free DMEM medium was added at a ratio of 2:1:1, and 12. mu.g polyetherimide (PEI, Polysciences, Inc.) was added. After mixing, standing for 16min, all the liquid was added to a six-well plate plated with HEK293T cells. After 6h of culture, the medium was discarded and fresh complete DMEM medium was added for culture. After 48h of transfection, the cell culture supernatant was collected and passed through a 0.45 μm filter (Millipore), to obtain a virus supernatant. Adding all the virus supernatant to the solution containing 1 × 10⁴HeK293T cells were plated in 6-well plates and incubated for 12h with polybrene (Sigma) at a final concentration of 4. mu.g/ml. The supernatant was then discarded and fresh complete DMEM medium was added. The obtained cell is the HEK293T-TIGIT cell.

HEK293T-TIGIT cells were diluted to 2X 10 with PBS⁶Perml, 100. mu.l/tube in 1.5ml EP tube, to which 10. mu.l/tube goat serum was added, and blocked at 4 ℃ for 30 min. TIGIT antibody was added at 1. mu.g/tube and incubated at 4 ℃ for 30 min. 1ml of PBS was added to the EP tube, centrifuged at 4 ℃ 3500 rpm. times.5 min, and the supernatant was discarded and washed with PBS. After centrifugation, the supernatant was discarded, the cells were resuspended in 100. mu.l/tube of PBS, and 0.1. mu.l/tube of Alexa-647-labeled goat anti-mouse antibody secondary antibody (Invitrogen) was added thereto, followed by incubation at 4 ℃ for 30min in the absence of light. Washed twice with PBS, centrifuged and the supernatant discarded. The cells were resuspended in 200. mu.l/tube PBS and assayed by flow cytometry, the results are shown in FIG. 2, further showing that the antibodies of the invention are capable of binding to TIGIT.

Example 3: in vitro binding affinity and kinetics experiments

The Biacore method is a well-known detection method for objectively detecting the mutual affinity and kinetics of proteins, and the characterization affinity and the binding kinetics of the TIGIT antibody are analyzed through Biacore T200.

TIGIT extracellular Fc fusion protein (TIGIT-Fc) was covalently linked to CM5(GE) chips using standard amino coupling methods. A series of concentration-graded TIGIT antibodies diluted in PBS were then injected into each cycle, and regenerated with 10mM NaOH solution. Following the kinetics of antigen-antibody binding for 3 minutes and dissociation for 10 minutes, the data were analyzed using the BIAevaluation software of GE in a 1: 1(Langmuir) binding model, and the values of ka (kon), KD (koff), and KD determined in this way are shown in Table 3 below.

TABLE 3TIGIT antibody affinities

Clone number	ka(1/Ms)	kd(1/s)	KD(M)
				1H6	1.545×10⁵	7.736×10^-4	5.007×10^-9
3A10	2.813×10⁴	9.031×10^-4	3.210×10^-8
				4A5	6.212×10⁵	2.318×10^-4	3.732×10^-10
4A9	6.725×10⁵	4.376×10^-3	6.507×10^-9
				4H5	5.222×10⁴	8.422×10^-4	1.613×10^-8
6A2	2.745E×10⁵	8.188×10^-4	2.983×10^-9
				687	1.571×10⁶	2.941×10^-4	1.872×10^-10
7F4	2.327×10⁵	2.707×10^-3	1.163×10^-9
				8E1	1.017×10⁵	2.640×10^-3	2.597×10^-9
8G3	2.434×10⁵	8.918×10^-4	3.664×10^-9
				9F4	3.990×10⁵	8.017×10^-4	2.009×10^-9
9G6	1.266×10⁵	3.404×10^-4	2.689×10^-9
				10F10	2.303×10⁶	3.581×10^-3	1.555×10^-9
11G4	7.577×10⁵	9.884×10^-4	1.305×10^-9
				12B7	3.671×10³	1.842×10^-4	5.019×10^-8
12C8	1.332×10⁵	7.673×10^-4	5.761×10^-9
				15E9	1.896×10⁶	7.022×10^-3	3.705×10^-9
16D6	2.080×10⁵	3.362×10^-3	1.617×10^-8
				16E10	1.959×10⁵	4.095×10^-4	2.091×10^-9

Example 4: screening for antibody blocking ability

TIGIT antibodies block the signaling pathway between TIGIT and its ligand by binding to the extracellular region of TIGIT, and flow cytometry experiments were used to detect the blocking of ligand CD155 binding by TIGIT antibodies.

HEK293T-TIGIT cells (supra) were diluted to 2X 10 with PBS⁶To a 1.5ml EP tube, 100. mu.l/tube of serum was added, and 10. mu.l/tube of mouse serum was blocked at 4 ℃ for 30 min. 4 μ g/tube of the Fc fusion protein (CD155-Fc, sequence shown in SEQ ID NO: 129) of the extracellular region of CD155 was added and incubated at 4 ℃ for 30 min. TIGIT antibodies (6A2 and 4A5) were added and incubated at 4 ℃ for 30 min. 1ml of PBS was added to the EP tube, centrifuged at 4 ℃ 3500 rpm. times.5 min, and the supernatant was discarded and washed with PBS. After centrifugation, the supernatant was discarded, the cells were resuspended in 100. mu.l/tube of PBS, and 1. mu.l/tube of 647-labeled mouse anti-human antibody secondary antibody (Biolegend) was added thereto, followed by incubation at 4 ℃ for 30min in the absence of light. Washed twice with PBS, centrifuged and the supernatant discarded. The cells were resuspended in 200. mu.l/tube PBS and assayed by flow cytometry, the results are shown in FIG. 3, and it can be seen that the antibodies of the invention are capable of blocking the binding of TIGIT to ligand CD 155.

Example 5: in vitro binding of NK cells

The test was used to detect the TIGIT antibodies of the invention and the native TIGIT binding activity expressed on the surface of human cells, which were human peripheral blood mononuclear cells.

Human peripheral blood mononuclear cells obtained by Ficoll density gradient centrifugation were diluted to 1X 10 with PBS⁷To a 1.5ml EP tube, 100. mu.l/tube of serum was added, and 10. mu.l/tube of mouse serum was blocked at 4 ℃ for 30 min. 1 μ g/tube Alexa-647 labeled TIGIT antibody (4A5) and other fluorescent antibodies (Brilliant Violet421TM-CD56, BioLegend; APC-CY7-CD3, BioLegend) were added and incubated at 4 ℃ for 30min protected from light. 1ml PBS was added to the EP tube, centrifuged at 3500 rpm. times.5 min at 4 ℃ and the supernatant was discarded and washed with PBS. After centrifugation, the supernatant was discarded, the cells were resuspended in 200. mu.l/tube PBS, and the detection was performed by flow cytometry, and the results are shown in FIG. 4, which shows that the antibody of the present invention can bind to human natural killer cells.

Example 6: antibody competition relation detection experiment

Competition for TIGIT antibodies obtained using flow cytometry methods.

HEK293T-TIGIT cells (supra) were diluted to 1X 10 with PBS⁶To a 1.5ml EP tube, 100. mu.l/tube of serum was added, and 10. mu.l/tube of mouse serum was blocked at 4 ℃ for 30 min. Add 1. mu.g/tube of non-fluorescently labeled TIGIT antibody and incubate at 4 ℃ for 30 min. Add 0.1. mu.g/tube of Andy flow 647(GeneCopoeia) labeled TIGIT antibody and incubate at 4 ℃ for 30 min. Washed twice with PBS, centrifuged and the supernatant discarded. The cells were resuspended in 200. mu.l/tube PBS and examined by flow cytometry. As shown in fig. 7, it can be seen that the antibodies of the present invention are divided into three groups according to their competitive relationship, 1H6, 3a10, 4a5, 4a9, 4H5, 6a2, 7F4, 8G3, 9F4, 10C1, 10F10, 12B7, 12C8, 15E9, 16D6, 16E10 are in one group and have a competitive relationship with each other; 6B7 and 16C11 are in a group, have competition relationship with each other and have no competition relationship with the group; 2B11, 8E1, 9G6 and 11G4 are in competition with each other and with the other two groups of antibodies.

Example 7: TIGIT antibody in vitro binding selectivity experiment

To test the specific binding activity of TIGIT antibodies (6a2, 4a5, and 9F4) to other proteins of the same family of TIGIT, human CD28, human PD-1, and human CTLA4 were used for binding assays. Meanwhile, in order to detect the difference of the TIGIT antibody to different species other than human, binding detection was also performed for TIGIT of mice.

Expression vectors containing human CD28(HG11524-CF, Chinesota), PD-1(HG10377-CF, Chinesota), CTLA4(HG11159-CF, Chinesota), and murine TIGIT (MG50939-CF, Chinesota) coding sequences were transfected into CHO-S cells (Thermo Fisher) using Lipofectamine 2000 reagent (Thermo Fisher) to overexpress human CD28, human PD-1, human CTLA4, and murine TIGIT on the surface of CHO-S cells. The method comprises the following specific steps: CHO-S cells were plated in six well plates, 1X 10⁶Cells/well, cultured overnight in DMEM medium without double antibody. Before transfection, the medium was discarded and 1mL of fresh DMEM medium without double antibody was added. Mu.g of plasmid, 3. mu.l of Lipofectamine 2000 liposomes were each added to an EP tube containing 250. mu.l of serum-free DMEM. After standing for 5min, the two tubes were mixed. Then, the mixture was allowed to stand for 20min, and the whole amount of the liquid was added to a six-well plate plated with CHO-S cells. After 5h of incubation, the medium was discarded and fresh complete DMEM medium was added. CHO-S cells were harvested 48h after transfection for detection.

CHO-S cells overexpressing the corresponding proteins were diluted to 2X 10 with PBS⁶Perml, 100. mu.l/tube in 1.5ml EP tube, to which 10. mu.l/tube goat serum was added, and blocked at 4 ℃ for 30 min. 2 ug/tube of TIGIT antibody was added and incubated at 4 ℃ for 30 min. 1ml of PBS was added to the EP tube, centrifuged at 4 ℃ 3500 rpm. times.5 min, and the supernatant was discarded and washed with PBS. After centrifugation, the supernatant was discarded, the cells were resuspended in 100. mu.l/tube of PBS, and 0.1. mu.l/tube of Alexa-647-labeled goat anti-mouse antibody secondary antibody (Invitrogen) was added thereto, followed by incubation at 4 ℃ for 30min in the absence of light. Washed twice with PBS, centrifuged and the supernatant discarded. Resuspending the cells in 200. mu.l/tube PBS and detecting by flow cytometry, as shown in FIGS. 5 and 6, it can be seen that the antibody of the present invention specifically binds to human TIGIT, but not to mouse TIGIT, and also does not bind to CD28, PD-1 and CTLA4 of the TIGIT superfamily.

Example 8: inhibition of tumor cell growth by TIGIT antibodies (in vitro killing)

In vitro killing experiment for detecting the effect of TIGIT antibody (6A2 and 4A5) in promoting the killing of tumor cells by human peripheral blood mononuclear cells. The tumor cells used were ovarian cancer cell HO8910 (Chinese academy of sciences cell Bank accession number TCHU24), human non-small cell lung cancer cell NCl-H358(ATCC accession number CRL5807), and human non-small cell lung cancer cell NCl-H1650(ATCC accession number CRL-5883), respectively.

(1) The tumor cells were washed twice with serum-free DMEM medium, centrifuged at 4 ℃ 3500rpm × 5min, and the supernatant was discarded. Resuspend with 1ml serum-free DMEM medium, add final concentration of 5. mu.M CFSE (succinimidyl ester), incubate 30min at 37 ℃.

(2) Add 200. mu.l of precooled fetal calf serum stop marker, centrifuge at 4 ℃ 3500 rpm. times.5 min and discard the supernatant. The cells were washed twice with complete RPMI-1640 medium containing 10% fetal bovine serum, and the supernatant was discarded. Resuspend with complete RPMI-1640 medium and count. Diluting with complete RPMI-1640 medium to 1.25 × 10⁵/ml。

(3) Human Peripheral Blood Mononuclear Cells (PBMC) obtained by Ficoll density gradient centrifugation were counted and diluted to a range of concentrations with complete RPMI-1640 medium containing 10% fetal bovine serum.

(4) Diluted PBMC were added to a 96-well round bottom plate in a volume of 100. mu.l/well.

(5) TIGIT antibody or control mouse IgG was diluted with complete RPMI-1640 medium and the diluted TIGIT antibody or mouse IgG was added to a 96-well round bottom plate in a volume of 20 μ l/well.

(6) The diluted tumor cells were added to a 96-well round bottom plate in a volume of 80. mu.l/well.

(7) Centrifugation was carried out at 1500rpm × 1min at 4 ℃ and incubation was carried out for 4h at 37 ℃.

(8) Add 7-AAD to a 96-well round bottom plate in a volume of 1. mu.l/well and mix well. All the liquid in the hole is transferred into a flow tube, and the detection is carried out by using a flow cytometer, and the result is shown in figure 8, so that the antibody disclosed by the invention can promote the anti-cancer function of the human peripheral blood mononuclear cell.

Example 9: experiment for inhibiting subcutaneous graft tumor

SW620 cells (ATCC No. CCL-227) were cultured in the same manner as described in 1X 10⁶Only inoculated in NOG mice (6-8 weeks,bred by the experimental animal center of the institute of Life sciences of the university of science and technology of China, the weight of which is about 20 g) under the right costal region, and the tumor grows to about 30mm on day 4³Thereafter, body weight, tumor over and under size were removed and mice were randomized by tumor volume into no treatment (control) group, PBMC + IgG group, PBMC +4A5mg/kg group and PBMC +6a2 group of 5 mice each. PBMC were prepared at 1.5X 10⁶One cell/mouse was injected into the tail vein and tail vein injection of antibody (6a2 and 4a5) was initiated on day 5, once on day 3 for a total of 8 doses. Tumor volumes were measured 4 times per week, mice weighed, and data recorded. Tumor volume (V) was calculated as: v1/2 × a × b²Wherein a and b represent length and width, respectively.

The results are shown in FIG. 9: the change of tumor volume after treatment is shown in the graph, and the antibody can well inhibit the growth of the tumor SW 620.

Example 10: mouse antibody humanization experiments

With reference to the light chain variable region sequence and the heavy chain variable region sequence of the 4a5 antibody, the humanized template best matching its non-CDR regions was selected. The CDR regions of the murine antibody were grafted onto the selected humanized template, and the CDR regions of the humanized template were replaced to give humanized antibody hu4A 5. Then, based on the three-dimensional structure of the murine antibody, the embedded residues, residues that interact directly with the CDR regions, and residues that have an important effect on the conformation of VL and VH were back-mutated to obtain a humanized antibody, in which the sequence of the heavy chain variable region of the humanized 4a5 antibody is as shown in SEQ ID NO:122, and the sequence of the humanized 4a5 antibody light chain variable region is shown in SEQ ID NO:123 and the heavy chain sequence is shown as' SEQ ID NO: 122-SEQ ID NO:124 'and the light chain sequence is as shown in' SEQ ID NO: 123-SEQ ID NO: 125' as shown. The murine variable region of the 4A5 antibody was fused to the constant region of human IgG4 to form the chimeric antibody ch4A5 (heavy chain sequence shown in SEQ ID NO:126 and light chain sequence shown in SEQ ID NO: 127).

Example 11: humanized antibody Activity data

ELISA binding experiment and ligand binding blocking experiment (same steps as in the previous examples) were performed in vitro on the chimeric antibody and the humanized antibody obtained in example 10, and the results are shown in FIG. 10 and FIG. 11, respectively; the results of the affinity kinetics experiments are shown in table 4 below, from which it can be seen that both the chimeric and humanized antibodies obtained in the present invention are capable of binding to TIGIT and blocking TIGIT binding to ligand CD 155.

TABLE 4 humanized 4A5 antibody affinity

	ka(1/Ms)	kd(1/s)	KD(M)
				ch4A5	2.610×10⁵	1.160×10^-4	4.450×10^-10
hu4A5	1.200×10⁵	3.693×10^-4	3.079×10^-9

From the above experimental results, it can be seen that the antibody obtained by the present invention can block the interaction between TIGIT and CD155 with high affinity; capable of binding with high specificity to TIGIT, but not to other CD28 family members (e.g., CD28, PD-1, and CTLA 4); and can activate immune cells, thereby killing tumor cells.

The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.

Sequence of

SEQ ID NO 1, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFTSYVMHWVKQKPGQGLEWIGYINPYNDDTNYNEKFKGKATLTSDKSSSTAYMELTSLTSEDSAVYFCARRLSVNSFPYWGQGTLVTVSA

2, heavy chain CDR1

GYTFTSYV

3, heavy chain CDR2

INPYNDDT

4, heavy chain CDR3

ARRLSVNSFPY

SEQ ID NO 5, light chain variable region

DIVMTQSHKFMSISVGDRVSISCKASQDVFTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDF TFTISSVQAEDLAVYYCQQHYSTPWTFGGGSNLEIK

6, light chain CDR1

QDVFTA

7, light chain CDR2

SAS

8, light chain CDR3

QQHYSTPWT

SEQ ID NO 9, heavy chain variable region

AVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTSYNPSLKSRISITR DTSKNQFFLQLNSVTTEDTATYYCARRGVGRGMDYWGQGTSVTVSS

10, heavy chain CDR1

GYSITSDYA

11, heavy chain CDR2

ISYSGST

12, heavy chain CDR3

ARRGVGRGMDY

SEQ ID NO 13, light chain variable region

DIVMTQSHKFMSTLVGDRVSITCKASQDIFTTVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDF TFTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLEIK

14, light chain CDR1

QDIFTT

SEQ ID NO 15, heavy chain variable region

DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTSYNPSLKSRISITR DTSKNQFFLQLNSVTTEDTATYYCARREVGQGFAHWGQGTLVTVSA

16, heavy chain CDR3

ARREVGQGFAH

SEQ ID NO 17, light chain variable region

DIVMTQSHKFMSTSVGDRVSITCKASQDLSTAVAWYQQKPGQSPKVLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLEIK

18, light chain CDR1

QDLSTA

SEQ ID NO 19, heavy chain variable region

LQESGPGLVKPSQSLSLTCTVTGSSITSDYAWNWIRQFPGNKLEWMGYITYSGRTTYNPSLKSRISITRDTS KNQFFLQLNSVTTEDTATYYCARWGLLRRYFDYWGQGTTLTVSS

20, heavy chain CDR1

GSSITSDYA

21, heavy chain CDR2

ITYSGRT

22, heavy chain CDR3

ARWGLLRRYFDY

SEQ ID NO 23, light chain variable region

DIVMTQSHKFMSTSVGDRVSITCKASQDVFTAVAWYQQKPGQSPKLLIYSASFRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPLTFGAGTKLELK

24, light chain CDR3

QQHYSTPLT

SEQ ID NO 25, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFTSFVMHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSADSAVYYCARRLSVSSFPYWGQGTLVTVSA

26, heavy chain CDR1

GYTFTSFV

27, heavy chain CDR2

INPYNDGT

28, heavy chain CDR3

ARRLSVSSFPY

SEQ ID NO 29, light chain variable region

DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPWTFGGGSKLEIK

30, light chain CDR1

QDVSTA

31, heavy chain variable region of SEQ ID NO

DVQLQESGPGLVKPSQSLSLTCTVTGYSITNDYAWNWIRQFPGNKLEWMGYISYSGSTSYNPSLKSRISIT RDTSKNQFFLQLNSVTTEDTATYYCARRGVGRTMDYWGQGTSVTVSS

32, heavy chain CDR1

GYSITNDYA

33, heavy chain CDR3

ARRGVGRTMDY

SEQ ID NO 34 light chain variable region

DIVMTQSHKFMSTSVGDRVSITCKASQDVSTVVGWYQQKPGQSPKSLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPWTFGGGTKLEIK

35, light chain CDR1

QDVSTV

SEQ ID NO 36, heavy chain variable region

EVQLQQSGPELVKPGTSVKMSCKASGYTFTSYVMHWVKQKPGQGLEWIGYINPYNDGAKYNEKFKGKVTLTSDKSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSA

37, heavy chain CDR2

INPYNDGA

38, heavy chain variable region

VQLKESGPGLVAPSQSLSITCTVSGFSLSTYSVHWIRQPPGEGLEWLGMIWGRGSTDYNSALRSRLSISKD NSKSQVFLKMNSLQTDDTAMYYCARNRGNLGYGGWFANWGQGTLVTVSA

39, heavy chain CDR1

GFSLSTYS

40, heavy chain CDR2

IWGRGST

41, heavy chain CDR3

ARNRGNLGYGGWFAN

SEQ ID NO 42 light chain variable region

DIVMTQSPSSLALSVGQKVTMSCKSSQSLLHSDNQKNYLAWYQQKPGQSPKLLVYFASTRESGVPDRFMGSGSGTDFTLTISGVQAEDLADYFCQQHYSTPWTFGGGTKLEIK

43 SEQ ID NO, light chain CDR1

QSLLHSDNQKNY

44, light chain CDR2

FAS

SEQ ID NO 45, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFTSYVMHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSA

SEQ ID NO 46 light chain variable region

DIVMTQSHKFMSTSVGDRVSITCKASQDVSNAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVFYCQHHYSTPPTFGAGTKLELK

47, light chain CDR1

QDVSNA

48, light chain CDR3

QHHYSTPPT

49, heavy chain variable region of SEQ ID NO

DVQLQESGPGLVKPSQSLSLTCTVTGFSITSDYVWNWIRQFPGNKLEWMGYISYSDTTMYNPSLKSRISIT RDTSKNQFFLQLNSVTTEDTATYFCARRDWAAWFVYWGQGTLVTVSA

50, heavy chain CDR1

GFSITSDYV

51, heavy chain CDR2

ISYSDTT

52, heavy chain CDR3

ARRDWAAWFVY

SEQ ID NO 53, light chain variable region

DIVMTQSHKFMSTSVGDRVTITCKASQDVSIAVAWYQQKPGQSPKLLVYSASFRYTGVPDRFTGSGSGTD FTFTISSVRAEDLAVYYCQQHYGTPWTFGGGTKLEIK

54, light chain CDR1

QDVSIA

55, light chain CDR3

QQHYGTPWT

56, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYIFTSYVMHWVKQKPGQGLEWIGYINPYNDGTKYYEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSA

57 heavy chain CDR1

GYIFTSYV

SEQ ID NO 58, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFSSYVMHWVKQKPGQGLEWIGYINPYNDGSKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSA

59, heavy chain CDR1

GYTFSSYV

60, heavy chain CDR2

INPYNDGS

61, heavy chain variable region of SEQ ID NO

DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTNYNPSLKSRISIT RDTSKNQFFLQLNSVTTEDTATYYCARRDWAAWFAYWGQGTLVTVSA

62, heavy chain CDR3

ARRDWAAWFAY

63, light chain variable region of SEQ ID NO

DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FSFTISSVQAEDLAVYYCQQQYSTPWTFGGGTKLEIK

64, light chain CDR3

QQQYSTPWT

65, heavy chain variable region

DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTSYNPSLKSRISISR DTSKNQFFLQLNSVTTEDTATYYCVRRDWAAWFAYWGQGTLVTVSA

66, heavy chain CDR3

VRRDWAAWFAY

67 variable region of light chain SEQ ID NO

SVPGDPLEIVSGAHGERVTMTCSASSSVSSSYFYWYQQKSGSSPKLWIYSISNLASGVPARFSGSGSGTSYS LKINSMEAEDAATYYCQQWSSNPPTFGGGTKLEIK

68, light chain CDR1

SSVSSSY

69, light chain CDR2

SIS

70, light chain CDR3

QQWSSNPPT

71, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFTSFVVHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDRSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSA

72, heavy chain variable region

EVQLQQSGPELVKPGASVKMSCKASGYTFTSYVLHWVKQKPGQGLEWIGYINPNNDVSKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARRLSVNSFPYWGQGTLVTVSS

73, heavy chain CDR2

INPNNDVS

74 light chain variable region of SEQ ID NO

DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSAPWTFGGGSKLEIK

75 SEQ ID NO, light chain CDR3

QQHYSAPWT

76 heavy chain variable region of SEQ ID NO

DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTSYNPSLKSRISITR DTSKNQFFLQLNSVTTEDTATYYCARRDWAAWFAHWGQGTLVTVSA

77 SEQ ID NO, heavy chain CDR3

ARRDWAAWFAH

78, heavy chain variable region

QVQLKESGPGLVAPSQSLSITCTVSGFSLSVYSIHWIRQPPGKGLEWLGMIWGGGITDYNSPLKSRLSISKD NSKSQVFLEMNGLQTDDTAIYYCARNRGNLGYGGWFADWGQGTLVTVS

79, heavy chain CDR1

GFSLSVYS

80, heavy chain CDR2

IWGGGIT

81 heavy chain CDR3 SEQ ID NO

ARNRGNLGYGGWFAD

82, light chain variable region of SEQ ID NO

DIVMTQSPSSLALSVGQKVTMTCKSSQSLLNGSNQVNYLAWYQQKPGQSPKLLVYFASTRESGVPDRFIG SGSGTDFTLTISSVQAEDLADYFCQQHYSTPWTFGGGTKLEIR

83, light chain CDR1

QSLLNGSNQVNY

84, light chain variable region of SEQ ID NO

DIVMTQSHKFMSTSVGDRVSITCKASQDVLTSVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPWTFGGGSKLEIK

85 SEQ ID NO, light chain CDR1

QDVLTS

86, heavy chain variable region

DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISYSGSTRYNPSLKSRISITR DTSKNQFFLQLNSVTTEDTATYYCARRELGQGFPYWGQGTLVTVSA

87, heavy chain CDR3

ARRELGQGFPY

88 SEQ ID NO, heavy chain variable region coding sequence

gaggtccagctacagcagtctggacctgagctggtgaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcactagctatgttatgcactggg tgaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatgatactaactacaatgagaagttcaaaggcaaggccacactgacttcagacaaatcctccagcacagcctacatggagctcaccagcctgacctctgaggactctgcggtctatttctgtgcaagaagactgtctgttaactcgtttccttactggggcca agggactctggtcactgtctctgcag

89 SEQ ID NO, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccatatcagtaggagacagggtcagcatctcctgcaaggccagtcaggatgtgtttactgctgtagcctggtatca acagaaaccaggacaatctcctaaactcctgatttactcggcatcctaccgatacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcac catcagcagtgtgcaggctgaggacctggcagtttattactgtcagcaacattatagtactccgtggacgttcggtggaggctccaacctggaaatcaaac

SEQ ID NO 90, heavy chain variable region coding sequence

gctgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactagctacaacccatctctcaaaagtcgaatctctatcacgcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagacgaggggtgggaagaggtatggactactgggg tcaaggaacctcagtcaccgtctcctcag

91 SEQ ID NO, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacattagtaggagacagggtcagcatcacctgcaaggccagtcaggatatatttactactgtagcctggtatca acagaaaccaggacaatctcctaaactactgatttattcggcatcctaccggtacactggagtccctgatcgtttcactggcagtggatctgggacggatttcactttcac catcagcagtgtgcaggctgaagacctggcagtttattattgtcagcaacattatagtactccgtggacgttcggtggaggcaccaagctggaaatcaaac

92, heavy chain variable region coding sequence

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactagctacaacccatctctcaaaagtcgaatctctatcactcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagaagggaagtgggacaggggtttgctcactgggg ccaagggactctggtcactgtctctgcag

93, light chain variable region coding sequence of SEQ ID NO

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatctgagtactgctgtagcctggtatc aacagaaaccaggacaatctcctaaagtactgatttactcggcatcctaccggtacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtactccgtggacgttcggtggaggcaccaagctggaaatcaaac

94 SEQ ID NO, heavy chain variable region coding sequence

gcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctcctcaatcaccagtgattatgcctggaactggatccggc agtttccaggaaacaaactggagtggatgggctacataacttacagtggtcgcactacctacaacccatctctcaaaagtcgaatctctatcactcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagatgggggttactacgtcgatactttgactactggggccaaggca ccactctcacagtctcctcag

SEQ ID NO 95, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtttttactgctgtggcctggtatca acagaaaccaggacaatctcctaaactactgatttactcggcatccttccggtacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcac catcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtactccgctcacgttcggtgctgggaccaagctggagctgaaac

96 SEQ ID NO, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcactagctttgttatgcactgggt gaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtactaagtacaatgagaagttcaaaggcaaggccacactgacttcagacaaatcctccagcacagcctacatggagctcagcagcctgacctctgcggactctgcggtctattactgtgcaagaaggctgtctgttagctcgtttccttactggggcc aagggactctggtcactgtctctgcag

97 of the light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtgagtactgctgtagcctggtatc aacagaaaccagggcaatctcctaaactcctgatttactcggcatcctaccgatacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtactccgtggacgttcggtggaggctccaagctggaaatcaaac

98 SEQ ID NO, variable region coding sequence of heavy chain

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccaatgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactagctacaacccatctctcaaaagtcgaatctctatcactcgagacacatccaagaaccaattcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagacgaggggtgggaagaactatggactactggggt caaggaacctcagtcaccgtctcctcag

99, light chain variable region coding sequence of SEQ ID NO

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtgagtactgttgtaggctggtatc aacagaaaccaggacaatctccaaaatcactgatttattcggcatcctaccggtacactggagtccctgatcgtttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtactccgtggacgttcggtggaggcaccaagctggaaatcaaac

100, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctgggacttcagtgaagatgtcctgcaaggcttctggctacacattcactagctatgttatgcactggg tgaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtgctaagtacaatgagaagttcaaaggcaaggtcacactgacttcagacaaatcctccagcacagcctacatggagctcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggcc aagggactctggtcactgtctctgcag

101, heavy chain variable region coding sequence of SEQ ID NO

gtgcagctgaaggagtcaggacctggcctggtggcaccctcacagagcctgtccatcacatgcactgtctctgggttctcattatccacatatagtgtacactggattcg ccagcctccaggagagggtctggagtggctgggaatgatctggggtcgtggaagcacagactataattcagctctcagatccagactgagcatcagcaaggacaac tccaagagtcaagttttcttaaaaatgaacagtctgcaaactgatgacacagccatgtactactgtgccagaaataggggtaatttgggttacggaggctggtttgctaac tggggccaagggactctggtcactgtctctgca

102, light chain variable region coding sequence

gacattgtgatgacacagtctccatcctccctggctttgtcagtaggacagaaggtcactatgagctgcaagtccagtcagagccttttacatagtgacaatcaaaagaa ctatttggcctggtaccagcagaaaccaggacagtctcctaaacttctggtatactttgcatccactagggaatctggggtccctgatcgcttcatgggcagtggatctgggacagatttcactcttaccatcagcggtgtgcaggctgaagacctggcagattacttctgtcagcaacattatagcactccgtggacgttcggtggaggcaccaagct ggaaatcaaac

103, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcactagctatgttatgcactggg tgaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtactaagtacaatgagaagttcaaagggaaggccacactgacttcag acaaatcctccagcacagcctacatggagctcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggc caagggactctggtcactgtctctgcag

104 SEQ ID NO, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtgagtaatgctgtagcctggtatc aacagaaaccaggacaatctcctaaactactgatttactcggcatcctaccggtacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagttttttactgtcagcaccattatagtactcctcccacgttcggtgctgggaccaagctggagctgaaac

105, heavy chain variable region coding sequence of SEQ ID NO

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggcttctcaatcaccagtgattacgtctggaactgg atccggcagtttccaggaaacaaactggagtggatgggctacataagctatagtgataccactatgtacaacccatctctcaaaagtcgaatctctatcactcgagacac atccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatatttctgtgcaagaagggactgggccgcctggtttgtttactggggccaag ggactctggtcactgtctctgcag

106 light chain variable region coding sequence of SEQ ID NO

gacattgtgatgacccagtctcacaaattcatgtccacatcagttggagacagggtcactatcacctgcaaggccagtcaggatgtgagtattgctgtagcctggtatca acagaaaccaggacaatctcctaaactactagtttactcggcatccttccggtacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcac catcagcagtgtgcgggctgaagacctggcagtttattactgtcagcaacattatggtactccgtggacgttcggtggaggcaccaagctggaaatcaaac

107 SEQ ID NO, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacatattcactagctatgttatgcactgggt gaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtactaagtactatgagaagttcaaaggcaaggccacactgacttcagacaaatcctccagcacagcctacatggagctcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggcca agggactctggtcactgtctctgcag

108 SEQ ID NO, heavy chain variable region coding sequence

gaggtccaactgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcagtagctatgttatgcactggg tgaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtagtaagtacaatgagaagttcaaaggcaaggccacactgacttcag acaaatcctccagcacagcctacatggagctcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggc caagggactctggtcactgtctctgcag

109, variable heavy chain coding sequence

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccgacagtttccaggaaacaagctggagtggatgggctacataagctacagtggtagcactaactacaacccatctctcaaaagtcgaatctctatcactcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagaagggactgggccgcctggtttgcttactggggcc aagggactctggtcactgtctctgcag

110, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgtgagtactgctgtagcctggtatc aacagaaaccaggacaatctcctaaactactgatttactcggcatcctaccggtacactggagtccctgatcgcttcactggcagtggatctgggacggatttcagtttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacaatatagtactccgtggacgttcggtggaggcaccaagctggaaatcaaac

111 SEQ ID NO, light chain variable region coding sequence

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactagctacaacccatctctcaaaagtcgaatctctatcagtcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgtaagaagggactgggccgcctggtttgcttactggggcc aagggactctggtcactgtctctgcag

112 SEQ ID NO, light chain variable region coding sequence

ctcggtacccggggatcctctagagattgtgtctggtgctcatggggaacgggtcaccatgacctgcagtgccagctcaagtgtgagttccagctacttttactggtacc agcagaagtcaggatcctccccaaaactctggatttatagcatatccaacctggcttctggagtcccagctcgcttcagtggcagtgggtctgggacctcttactctctca aaatcaacagcatggaggctgaagatgctgccacttattactgccagcagtggagtagtaacccaccgacgttcggtggaggcaccaagctggaaatcaaac

113, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcactagctttgttgtgcactggg tgaagcagaagcctgggcagggccttgagtggattggatatattaatccttacaatgatggtactaagtacaatgagaagttcaaaggcaaggccacactgacttcagacaggtcctccagcacagcctacatggagctcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggcc aagggactctggtcactgtctctgcag

114, heavy chain variable region coding sequence

gaggtccagctgcagcagtctggacctgagctggtaaagcctggggcttcagtgaagatgtcctgcaaggcttctggatacacattcactagctatgttttgcactgggt gaagcagaagcctgggcagggccttgagtggattggatacattaatcctaacaatgatgttagtaagtacaatgagaagttcaaaggcaaggccacactgacttcagacaaatcctccagcacagcctacatggaactcagcagcctgacctctgaggactctgcggtctattactgtgcaagaaggctgtctgttaactcgtttccttactggggcc aagggactctggtcactgtctcttcag

115 SEQ ID NO, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtcggagacagggtcagcatcacctgcaaggccagtcaggatgtgagtactgctgtagcctggtatc aacagaaaccagggcaatctcctaaactcctgatttactcggcatcctaccgatacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtgctccgtggacgttcggtggaggctccaagctggaaatcaaac

116, heavy chain variable region coding sequence

gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactagctacaacccatctctcaaaagtcgaatttctatcactcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaagacacagccacatattactgtgcaagaagggactgggccgcctggtttgctcactggggcc aagggactctggtcactgtctctgcag

117, heavy chain variable region coding sequence

caggtgcagctgaaggagtcaggacctggcctggtggcaccctcacagagcctgtccatcacatgcactgtctctgggttctcattatccgtatatagtatacactggat tcgccagcctccaggaaagggtctggagtggctgggaatgatctggggtggtggaatcacagactataattcacctctcaaatccagactgagcatcagcaaggaca actccaagagccaagttttcttagaaatgaacggtctgcaaactgatgacacagccatctactactgtgccagaaataggggtaatttgggttacggaggctggtttgct gactggggccaagggactctggtcactgtctctgcag

118, light chain variable region coding sequence of SEQ ID NO

gacattgtgatgacacagtctccatcctccctggctctgtcagtaggacagaaggtcactatgacctgcaagtccagtcagagccttttaaatggtagcaatcaagtgaa ctatttggcctggtaccaacagaaaccaggacagtctcctaaacttctggtgtactttgcgtccactagggaatctggggtccctgatcgcttcataggcagtggatctgggacagatttcactcttaccatcagcagtgtgcaggctgaagacctggcagattacttctgtcagcaacattatagcactccgtggacgttcggtggaggcaccaagct ggaaatcagac

119 SEQ ID NO, light chain variable region coding sequence

gacattgtgatgacccagtctcacaaattcatgtccacatcagtaggagacagggtcagcatcacctgcaaggccagtcaggatgttcttacatctgtagcctggtatca acagaaaccagggcaatctcctaaactcctgatttactcggcatcctaccgatacactggagtccctgatcgcttcactggcagtggatctgggacggatttcactttcaccatcagcagtgtgcaggctgaagacctggcagtttattactgtcagcaacattatagtactccgtggacgttcggtggaggctccaagctggaaatcaaac

120, heavy chain variable region coding sequence of SEQ ID NO

Gatgtgcagcttcaggagtcgggacctggcctggtgaaaccttctcagtctctgtccctcacctgcactgtcactggctactcaatcaccagtgattatgcctggaactg gatccggcagtttccaggaaacaaactggagtggatgggctacataagctacagtggtagcactaggtacaacccatctctcaaaagtcgaatctctatcactcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagaagggaactgggacaggggtttccttactggggc caagggactctggtcactgtctctgcag

121 in SEQ ID NO, recombinant TIGIT extracellular region Fc fusion protein

MMTGTIETTGNISAEKGGSIILQCHLSSTTAQVTQVNWEQQDQLLAICNADLGWHISPSFKDRVAPGPGLG LTLQSLTVNDTGEYFCIYHTYPDGTYTGRIFLEVLELEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

122, humanized 4A5 antibody heavy chain variable region sequence

QVQLQESGPGLVKPSQTLSLTCTVTGSSITSDYAWNWIRQFPGKKLEWMGYITYSGRTTYNPSLKSRITISR DTSKNQFSLKLSSVTAADTATYYCARWGLLRRYFDYWGQGTLLTVSS

123 SEQ ID NO, humanized 4A5 antibody light chain variable region sequence

DIVMTQSPSSMSTSVGDRVTITCRASQDVFTAVAWYQQKPGKSPKLLIYSASFRYTGVPDRFSGSGSGTDF TFTISSVQAEDFATYYCQQHYSTPLTFGAGTKLELK

124, human IgG4 heavy chain constant region sequence

ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPS SIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

125 SEQ ID NO, human IgG4 kappa chain constant region sequence

RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

126, 4A5 chimeric antibody heavy chain sequence

QVQLQESGPGLVKPSQSLSLTCTVTGSSITSDYAWNWIRQFPGNKLEWMGYITYSGRTTYNPSLKSRISITR DTSKNQFFLQLNSVTTEDTATYYCARWGLLRRYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTK VDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPP SQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

Light chain sequence of the chimeric antibody of SEQ ID NO 127, 4A5

DIVMTQSHKFMSTSVGDRVSITCKASQDVFTAVAWYQQKPGQSPKLLIYSASFRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYSTPLTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GEC

128, TIGIT protein coding sequence

atgcgctggtgtctcctcctgatctgggcccaggggctgaggcaggctcccctcgcctcaggaatgatgacaggcacaatagaaacaacggggaacatttctgcag agaaaggtggctctatcatcttacaatgtcacctctcctccaccacggcacaagtgacccaggtcaactgggagcagcaggaccagcttctggccatttgtaatgctga cttggggtggcacatctccccatccttcaaggatcgagtggccccaggtcccggcctgggcctcaccctccagtcgctgaccgtgaacgatacaggggagtacttct gcatctatcacacctaccctgatgggacgtacactgggagaatcttcctggaggtcctagaaagctcagtggctgagcacggtgccaggttccagattccattgcttgg agccatggccgcgacgctggtggtcatctgcacagcagtcatcgtggtggtcgcgttgactagaaagaagaaagccctcagaatccattctgtggaaggtgacctca ggagaaaatcagctggacaggaggaatggagccccagtgctccctcacccccaggaagctgtgtccaggcagaagctgcacctgctgggctctgtggagagcag cggggagaggactgtgccgagctgcatgactacttcaatgtcctgagttacagaagcctgggtaactgcagcttcttcacagagactggttag

129, CD155-Fc protein sequence

WPPPGTGDVVVQAPTQVPGFLGDSVTLPCYLQVPNMEVTHVSQLTWARHGESGSMAVFHQTQGPSYSESKRLEFVAARLGAELRNASLRMFGLRVEDEGNYTCLFVTFPQGSRSVDIWLRVLAKPQNTAEVQKVQLT GEPVPMARCVSTGGRPPAQITWHSDLGGMPNTSQVPGFLSGTVTVTSLWILVPSSQVDGKNVTCKVEHES FEKPQLLTVNLTVYYPPEVSISGYDNNWYLGQNEATLTCDARSNPEPTGYNWSTTMGPLPPFAVAQGAQL LIRPVDKPINTTLICNVTNALGARQAELTVQVKEGPPSEHSGLEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK。

Sequence listing

<110> Hefei Ruida Immunity drug research institute Co., Ltd

<120> antibody capable of binding to TIGIT or antigen-binding fragment thereof, and use

<141>2018-09-25

<160>129

<170>SIPOSequenceListing 1.0

<210>1

<211>118

<212>PRT

<213> heavy chain variable region

<400>1

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Asp Thr Asn Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>2

<211>8

<212>PRT

<213> heavy chain CDR1

<400>2

Gly Tyr Thr Phe Thr Ser Tyr Val

1 5

<210>3

<211>8

<212>PRT

<213> heavy chain CDR2

<400>3

Ile Asn Pro Tyr Asn Asp Asp Thr

1 5

<210>4

<211>11

<212>PRT

<213> heavy chain CDR3

<400>4

Ala Arg Arg Leu Ser Val Asn Ser Phe Pro Tyr

1 5 10

<210>5

<211>107

<212>PRT

<213> light chain variable region

<400>5

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Ile Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Ser Cys Lys Ala Ser Gln Asp Val Phe Thr Ala

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Ser Asn Leu Glu Ile Lys

100 105

<210>6

<211>6

<212>PRT

<213> light chain CDR1

<400>6

Gln Asp Val Phe Thr Ala

1 5

<210>7

<211>3

<212>PRT

<213> light chain CDR2

<400>7

Ser Ala Ser

1

<210>8

<211>9

<212>PRT

<213> light chain CDR3

<400>8

Gln Gln His Tyr Ser Thr Pro Trp Thr

1 5

<210>9

<211>118

<212>PRT

<213> heavy chain variable region

<400>9

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Ser Val Thr Val Ser Ser

115

<210>10

<211>9

<212>PRT

<213> heavy chain CDR1

<400>10

Gly Tyr Ser Ile Thr Ser Asp Tyr Ala

1 5

<210>11

<211>7

<212>PRT

<213> heavy chain CDR2

<400>11

Ile Ser Tyr Ser Gly Ser Thr

1 5

<210>12

<211>11

<212>PRT

<213> heavy chain CDR3

<400>12

Ala Arg Arg Gly Val Gly Arg Gly Met Asp Tyr

1 5 10

<210>13

<211>107

<212>PRT

<213> light chain variable region

<400>13

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Leu Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Ile Phe Thr Thr

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>14

<211>6

<212>PRT

<213> light chain CDR1

<400>14

Gln Asp Ile Phe Thr Thr

1 5

<210>15

<211>118

<212>PRT

<213> heavy chain variable region

<400>15

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Arg Glu Val Gly Gln Gly Phe Ala His Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala

115

<210>16

<211>11

<212>PRT

<213> heavy chain CDR3

<400>16

Ala Arg Arg Glu Val Gly Gln Gly Phe Ala His

1 5 10

<210>17

<211>107

<212>PRT

<213> light chain variable region

<400>17

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Leu Ser Thr Ala

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>18

<211>6

<212>PRT

<213> light chain CDR1

<400>18

Gln Asp Leu Ser Thr Ala

1 5

<210>19

<211>116

<212>PRT

<213> heavy chain variable region

<400>19

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

1 5 10 15

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

20 25 30

Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp Met Gly Tyr

35 40 45

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

50 55 60

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

65 70 75 80

Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala Arg Trp

85 90 95

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

100 105 110

Thr Val Ser Ser

115

<210>20

<211>9

<212>PRT

<213> heavy chain CDR1

<400>20

Gly Ser Ser Ile Thr Ser Asp Tyr Ala

1 5

<210>21

<211>7

<212>PRT

<213> heavy chain CDR2

<400>21

Ile Thr Tyr Ser Gly Arg Thr

1 5

<210>22

<211>12

<212>PRT

<213> heavy chain CDR3

<400>22

Ala Arg Trp Gly Leu Leu Arg Arg Tyr Phe Asp Tyr

1 5 10

<210>23

<211>107

<212>PRT

<213> light chain variable region

<400>23

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Phe Thr Ala

20 25 30

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

35 4045

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

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

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210>24

<211>9

<212>PRT

<213> light chain CDR3

<400>24

Gln Gln His Tyr Ser Thr Pro Leu Thr

1 5

<210>25

<211>118

<212>PRT

<213> heavy chain variable region

<400>25

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Phe

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>26

<211>8

<212>PRT

<213> heavy chain CDR1

<400>26

Gly Tyr Thr Phe Thr Ser Phe Val

1 5

<210>27

<211>8

<212>PRT

<213> heavy chain CDR2

<400>27

Ile Asn Pro Tyr Asn Asp Gly Thr

1 5

<210>28

<211>11

<212>PRT

<213> heavy chain CDR3

<400>28

Ala Arg Arg Leu Ser Val Ser Ser Phe Pro Tyr

1 5 10

<210>29

<211>107

<212>PRT

<213> light chain variable region

<400>29

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Ser Lys Leu Glu Ile Lys

100 105

<210>30

<211>6

<212>PRT

<213> light chain CDR1

<400>30

Gln Asp Val Ser Thr Ala

1 5

<210>31

<211>6

<212>PRT

<213> heavy chain variable region

<400>31

Gln Asp Val Ser Thr Ala

1 5

<210>32

<211>9

<212>PRT

<213> heavy chain CDR1

<400>32

Gly Tyr Ser Ile Thr Asn Asp Tyr Ala

1 5

<210>33

<211>11

<212>PRT

<213> heavy chain CDR3

<400>33

Ala Arg Arg Gly Val Gly Arg Thr Met Asp Tyr

1 5 10

<210>34

<211>107

<212>PRT

<213> light chain variable region

<400>34

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

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

20 25 30

Val Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ser Leu Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>35

<211>6

<212>PRT

<213> light chain CDR1

<400>35

Gln Asp Val Ser Thr Val

1 5

<210>36

<211>118

<212>PRT

<213> heavy chain variable region

<400>36

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Ala Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>37

<211>8

<212>PRT

<213> heavy chain CDR2

<400>37

Ile Asn Pro Tyr Asn Asp Gly Ala

1 5

<210>38

<211>120

<212>PRT

<213> heavy chain variable region

<400>38

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

1 5 10 15

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

20 25 30

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

35 40 45

Met Ile Trp Gly Arg Gly Ser Thr Asp Tyr Asn Ser Ala Leu Arg Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Gly Thr Leu Val Thr Val Ser Ala

115 120

<210>39

<211>8

<212>PRT

<213> heavy chain CDR1

<400>39

Gly Phe Ser Leu Ser Thr Tyr Ser

1 5

<210>40

<211>7

<212>PRT

<213> heavy chain CDR2

<400>40

Ile Trp Gly Arg Gly Ser Thr

1 5

<210>41

<211>7

<212>PRT

<213> heavy chain CDR3

<400>41

Ile Trp Gly Arg Gly Ser Thr

1 5

<210>42

<211>113

<212>PRT

<213> light chain variable region

<400>42

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

1 5 10 15

Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu His Ser

20 25 30

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

35 40 45

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

50 55 60

Pro Asp Arg Phe Met Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Gly Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln

85 90 95

His Tyr Ser Thr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

100 105 110

Lys

<210>43

<211>12

<212>PRT

<213> light chain CDR1

<400>43

Gln Ser Leu Leu His Ser Asp Asn Gln Lys Asn Tyr

1 5 10

<210>44

<211>3

<212>PRT

<213> light chain CDR2

<400>44

Phe Ala Ser

1

<210>45

<211>118

<212>PRT

<213> heavy chain variable region

<400>45

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

1 510 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>46

<211>107

<212>PRT

<213> light chain variable region

<400>46

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Asn Ala

2025 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Phe Tyr Cys Gln His His Tyr Ser Thr Pro Pro

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210>47

<211>6

<212>PRT

<213> light chain CDR1

<400>47

Gln Asp Val Ser Asn Ala

1 5

<210>48

<211>9

<212>PRT

<213> light chain CDR3

<400>48

Gln His His Tyr Ser Thr Pro Pro Thr

1 5

<210>49

<211>118

<212>PRT

<213> heavy chain variable region

<400>49

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

1 5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Phe Ser Ile Thr Ser Asp

20 25 30

Tyr Val Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

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

50 55 60

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

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>50

<211>9

<212>PRT

<213> heavy chain CDR1

<400>50

Gly Phe Ser Ile Thr Ser Asp Tyr Val

1 5

<210>51

<211>7

<212>PRT

<213> heavy chain CDR2

<400>51

Ile Ser Tyr Ser Asp Thr Thr

1 5

<210>52

<211>11

<212>PRT

<213> heavy chain CDR3

<400>52

Ala Arg Arg Asp Trp Ala Ala Trp Phe Val Tyr

1 5 10

<210>53

<211>107

<212>PRT

<213> light chain variable region

<400>53

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

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

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Val

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Gly Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>54

<211>6

<212>PRT

<213> light chain CDR1

<400>54

Gln Asp Val Ser Ile Ala

1 5

<210>55

<211>9

<212>PRT

<213> light chain CDR3

<400>55

Gln Gln His Tyr Gly Thr Pro Trp Thr

1 5

<210>56

<211>118

<212>PRT

<213> heavy chain variable region

<400>56

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Ser Tyr

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Tyr Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>57

<211>8

<212>PRT

<213> heavy chain CDR1

<400>57

Gly Tyr Ile Phe Thr Ser Tyr Val

1 5

<210>58

<211>118

<212>PRT

<213> heavy chain variable region

<400>58

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Tyr

20 25 30

Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Ser Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>59

<211>8

<212>PRT

<213> heavy chain CDR1

<400>59

Gly Tyr Thr Phe Ser Ser Tyr Val

1 5

<210>60

<211>8

<212>PRT

<213> heavy chain CDR2

<400>60

Ile Asn Pro Tyr Asn Asp Gly Ser

1 5

<210>61

<211>118

<212>PRT

<213> heavy chain variable region

<400>61

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>62

<211>11

<212>PRT

<213> heavy chain CDR3

<400>62

Ala Arg Arg Asp Trp Ala Ala Trp Phe Ala Tyr

1 5 10

<210>63

<211>107

<212>PRT

<213> light chain variable region

<400>63

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Ser Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

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

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>64

<211>9

<212>PRT

<213> light chain CDR3

<400>64

Gln Gln Gln Tyr Ser Thr Pro Trp Thr

1 5

<210>65

<211>118

<212>PRT

<213> heavy chain variable region

<400>65

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>66

<211>11

<212>PRT

<213> heavy chain CDR3

<400>66

Val Arg Arg Asp Trp Ala Ala Trp Phe Ala Tyr

1 5 10

<210>67

<211>107

<212>PRT

<213> light chain variable region

<400>67

Ser Val Pro Gly Asp Pro Leu Glu Ile Val Ser Gly Ala His Gly Glu

1 5 10 15

Arg Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Ser Ser Tyr

20 25 30

Phe Tyr Trp Tyr Gln Gln Lys Ser Gly Ser Ser Pro Lys Leu Trp Ile

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Ser Tyr Ser Leu Lys Ile Asn Ser Met Glu Ala

65 70 75 80

Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210>68

<211>7

<212>PRT

<213> light chain CDR1

<400>68

Ser Ser Val Ser Ser Ser Tyr

1 5

<210>69

<211>3

<212>PRT

<213> light chain CDR2

<400>69

Ser Ile Ser

1

<210>70

<211>9

<212>PRT

<213> light chain CDR3

<400>70

Gln Gln Trp Ser Ser Asn Pro Pro Thr

1 5

<210>71

<211>118

<212>PRT

<213> heavy chain variable region

<400>71

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Phe

20 25 30

Val Val His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>72

<211>118

<212>PRT

<213> heavy chain variable region

<400>72

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

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Val Leu His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Asn Asn Asp Val Ser Lys Tyr Asn Glu Lys Phe

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ser

115

<210>73

<211>8

<212>PRT

<213> heavy chain CDR2

<400>73

Ile Asn Pro Asn Asn Asp Val Ser

1 5

<210>74

<211>107

<212>PRT

<213> light chain variable region

<400>74

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Ala Pro Trp

85 90 95

Thr Phe Gly Gly Gly Ser Lys Leu Glu Ile Lys

100 105

<210>75

<211>9

<212>PRT

<213> light chain CDR3

<400>75

Gln Gln His Tyr Ser Ala Pro Trp Thr

1 5

<210>76

<211>118

<212>PRT

<213> heavy chain variable region

<400>76

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Arg Asp Trp Ala Ala Trp Phe Ala His Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala

115

<210>77

<211>11

<212>PRT

<213> heavy chain CDR3

<400>77

Ala Arg Arg Asp Trp Ala Ala Trp Phe Ala His

1 510

<210>78

<211>120

<212>PRT

<213> heavy chain variable region

<400>78

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Met Asn Gly Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Arg Asn Arg Gly Asn Leu Gly Tyr Gly Gly Trp Phe Ala Asp Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser

115 120

<210>79

<211>8

<212>PRT

<213> heavy chain CDR1

<400>79

Gly Phe Ser Leu Ser Val Tyr Ser

1 5

<210>80

<211>7

<212>PRT

<213> heavy chain CDR2

<400>80

Ile Trp Gly Gly Gly Ile Thr

1 5

<210>81

<211>15

<212>PRT

<213> heavy chain CDR3

<400>81

Ala Arg Asn Arg Gly Asn Leu Gly Tyr Gly Gly Trp Phe Ala Asp

1 5 10 15

<210>82

<211>113

<212>PRT

<213> light chain variable region

<400>82

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

1 5 10 15

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

20 25 30

Ser Asn Gln Val Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

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

50 55 60

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

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln

85 90 95

His Tyr Ser Thr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

100 105 110

Arg

<210>83

<211>12

<212>PRT

<213> light chain CDR1

<400>83

Gln Ser Leu Leu Asn Gly Ser Asn Gln Val Asn Tyr

1 5 10

<210>84

<211>107

<212>PRT

<213> light chain variable region

<400>84

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Leu Thr Ser

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Ser Lys Leu Glu Ile Lys

100 105

<210>85

<211>6

<212>PRT

<213> light chain CDR1

<400>85

Gln Asp Val Leu Thr Ser

1 5

<210>86

<211>118

<212>PRT

<213> heavy chain variable region

<400>86

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

1 5 10 15

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

20 25 30

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

35 40 45

Met Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Arg Tyr Asn Pro Ser Leu

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Leu Val Thr Val Ser Ala

115

<210>87

<211>11

<212>PRT

<213> heavy chain CDR3

<400>87

Ala Arg Arg Glu Leu Gly Gln Gly Phe Pro Tyr

1 5 10

<210>88

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>88

gaggtccagc tacagcagtc tggacctgag ctggtgaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatga tactaactac 180

aatgagaagt tcaaaggcaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca ccagcctgac ctctgaggac tctgcggtct atttctgtgc aagaagactg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>89

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>89

gacattgtga tgacccagtc tcacaaattc atgtccatat cagtaggaga cagggtcagc 60

atctcctgca aggccagtca ggatgtgttt actgctgtag cctggtatca acagaaacca 120

ggacaatctc ctaaactcct gatttactcg gcatcctacc gatacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaggacctgg cagtttatta ctgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggctccaacc tggaaatcaa ac 322

<210>90

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>90

gctgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct acagtggtag cactagctac 180

aacccatctc tcaaaagtcg aatctctatc acgcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagacgaggg 300

gtgggaagag gtatggacta ctggggtcaa ggaacctcag tcaccgtctc ctcag 355

<210>91

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>91

gacattgtga tgacccagtc tcacaaattc atgtccacat tagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatatattt actactgtag cctggtatca acagaaacca 120

ggacaatctc ctaaactact gatttattcg gcatcctacc ggtacactgg agtccctgat 180

cgtttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ttgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggcaccaagc tggaaatcaa ac 322

<210>92

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>92

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct acagtggtag cactagctac 180

aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagaagggaa 300

gtgggacagg ggtttgctca ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>93

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>93

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatctgagt actgctgtag cctggtatca acagaaacca 120

ggacaatctc ctaaagtact gatttactcg gcatcctacc ggtacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggcaccaagc tggaaatcaa ac 322

<210>94

<211>350

<212>DNA

<213> heavy chain variable region coding sequence

<400>94

gcttcaggag tcgggacctg gcctggtgaa accttctcag tctctgtccc tcacctgcac 60

tgtcactggc tcctcaatca ccagtgatta tgcctggaac tggatccggc agtttccagg 120

aaacaaactg gagtggatgg gctacataac ttacagtggt cgcactacct acaacccatc 180

tctcaaaagt cgaatctcta tcactcgaga cacatccaag aaccagttct tcctgcagtt 240

gaattctgtg actactgagg acacagccac atattactgt gcaagatggg ggttactacg 300

tcgatacttt gactactggg gccaaggcac cactctcaca gtctcctcag 350

<210>95

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>95

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgttttt actgctgtgg cctggtatca acagaaacca 120

ggacaatctc ctaaactact gatttactcg gcatccttcc ggtacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagta ctccgctcac gttcggtgct 300

gggaccaagc tggagctgaa ac 322

<210>96

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>96

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctttgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tactaagtac 180

aatgagaagt tcaaaggcaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgcggac tctgcggtct attactgtgc aagaaggctg 300

tctgttagct cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>97

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>97

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acagaaacca 120

gggcaatctc ctaaactcct gatttactcg gcatcctacc gatacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggctccaagc tggaaatcaa ac 322

<210>98

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>98

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc aatgattatg cctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct acagtggtag cactagctac 180

aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaagaa ccaattcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagacgaggg 300

gtgggaagaa ctatggacta ctggggtcaa ggaacctcag tcaccgtctc ctcag 355

<210>99

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>99

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgtgagt actgttgtag gctggtatca acagaaacca 120

ggacaatctc caaaatcact gatttattcg gcatcctacc ggtacactgg agtccctgat 180

cgtttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggcaccaagc tggaaatcaa ac 322

<210>100

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>100

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctgggacttc agtgaagatg 60

tcctgcaagg cttctggcta cacattcact agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tgctaagtac 180

aatgagaagt tcaaaggcaa ggtcacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>101

<211>360

<212>DNA

<213> heavy chain variable region coding sequence

<400>101

gtgcagctga aggagtcagg acctggcctg gtggcaccct cacagagcct gtccatcaca 60

tgcactgtct ctgggttctc attatccaca tatagtgtac actggattcg ccagcctcca 120

ggagagggtc tggagtggct gggaatgatc tggggtcgtg gaagcacaga ctataattca 180

gctctcagat ccagactgag catcagcaag gacaactcca agagtcaagt tttcttaaaa 240

atgaacagtc tgcaaactga tgacacagcc atgtactact gtgccagaaa taggggtaat 300

ttgggttacg gaggctggtt tgctaactgg ggccaaggga ctctggtcac tgtctctgca 360

<210>102

<211>355

<212>DNA

<213> light chain variable region coding sequence

<400>102

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tactaagtac 180

aatgagaagt tcaaagggaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>103

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>103

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tactaagtac 180

aatgagaagt tcaaagggaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>104

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>104

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgtgagt aatgctgtag cctggtatca acagaaacca 120

ggacaatctc ctaaactact gatttactcg gcatcctacc ggtacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttttta ctgtcagcac cattatagta ctcctcccac gttcggtgct 300

gggaccaagc tggagctgaa ac 322

<210>105

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>105

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggctt ctcaatcacc agtgattacg tctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct atagtgatac cactatgtac 180

aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat atttctgtgc aagaagggac 300

tgggccgcct ggtttgttta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>106

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>106

gacattgtga tgacccagtc tcacaaattc atgtccacat cagttggaga cagggtcact 60

atcacctgca aggccagtca ggatgtgagt attgctgtag cctggtatca acagaaacca 120

ggacaatctc ctaaactact agtttactcg gcatccttcc ggtacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcgggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatggta ctccgtggac gttcggtgga 300

ggcaccaagc tggaaatcaa ac 322

<210>107

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>107

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata catattcact agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tactaagtac 180

tatgagaagt tcaaaggcaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>108

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>108

gaggtccaac tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcagt agctatgtta tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tagtaagtac 180

aatgagaagt tcaaaggcaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>109

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>109

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccgacag 120

tttccaggaa acaagctgga gtggatgggc tacataagct acagtggtag cactaactac 180

aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagaagggac 300

tgggccgcct ggtttgctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>110

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>110

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acagaaacca 120

ggacaatctc ctaaactact gatttactcg gcatcctacc ggtacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcagtttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa caatatagta ctccgtggac gttcggtgga 300

ggcaccaagc tggaaatcaa ac 322

<210>111

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>111

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct acagtggtag cactagctac 180

aacccatctc tcaaaagtcg aatctctatc agtcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgt aagaagggac 300

tgggccgcct ggtttgctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>112

<211>323

<212>DNA

<213> light chain variable region coding sequence

<400>112

ctcggtaccc ggggatcctc tagagattgt gtctggtgct catggggaac gggtcaccat 60

gacctgcagt gccagctcaa gtgtgagttc cagctacttt tactggtacc agcagaagtc 120

aggatcctcc ccaaaactct ggatttatag catatccaac ctggcttctg gagtcccagc 180

tcgcttcagt ggcagtgggt ctgggacctc ttactctctc aaaatcaaca gcatggaggc 240

tgaagatgct gccacttatt actgccagca gtggagtagt aacccaccga cgttcggtgg 300

aggcaccaag ctggaaatca aac 323

<210>113

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>113

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctttgttg tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatat attaatcctt acaatgatgg tactaagtac 180

aatgagaagt tcaaaggcaa ggccacactg acttcagaca ggtcctccag cacagcctac 240

atggagctca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>114

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>114

gaggtccagc tgcagcagtc tggacctgag ctggtaaagc ctggggcttc agtgaagatg 60

tcctgcaagg cttctggata cacattcact agctatgttt tgcactgggt gaagcagaag 120

cctgggcagg gccttgagtg gattggatac attaatccta acaatgatgt tagtaagtac 180

aatgagaagt tcaaaggcaa ggccacactg acttcagaca aatcctccag cacagcctac 240

atggaactca gcagcctgac ctctgaggac tctgcggtct attactgtgc aagaaggctg 300

tctgttaact cgtttcctta ctggggccaa gggactctgg tcactgtctc ttcag 355

<210>115

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>115

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtcggaga cagggtcagc 60

atcacctgca aggccagtca ggatgtgagt actgctgtag cctggtatca acagaaacca 120

gggcaatctc ctaaactcct gatttactcg gcatcctacc gatacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagtg ctccgtggac gttcggtgga 300

ggctccaagc tggaaatcaa ac 322

<210>116

<211>355

<212>DNA

<213> heavy chain variable region coding sequence

<400>116

gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60

acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120

tttccaggaa acaaactgga gtggatgggc tacataagct acagtggtag cactagctac 180

aacccatctc tcaaaagtcg aatttctatc actcgagaca catccaagaa ccagttcttc 240

ctgcagttga attctgtgac tactgaagac acagccacat attactgtgc aagaagggac 300

tgggccgcct ggtttgctca ctggggccaa gggactctgg tcactgtctc tgcag 355

<210>117

<211>364

<212>DNA

<213> heavy chain variable region coding sequence

<400>117

caggtgcagc tgaaggagtc aggacctggc ctggtggcac cctcacagag cctgtccatc 60

acatgcactg tctctgggtt ctcattatcc gtatatagta tacactggat tcgccagcct 120

ccaggaaagg gtctggagtg gctgggaatg atctggggtg gtggaatcac agactataat 180

tcacctctca aatccagact gagcatcagc aaggacaact ccaagagcca agttttctta 240

gaaatgaacg gtctgcaaac tgatgacaca gccatctact actgtgccag aaataggggt 300

aatttgggtt acggaggctg gtttgctgac tggggccaag ggactctggt cactgtctct 360

gcag 364

<210>118

<211>340

<212>DNA

<213> light chain variable region coding sequence

<400>118

gacattgtga tgacacagtc tccatcctcc ctggctctgt cagtaggaca gaaggtcact 60

atgacctgca agtccagtca gagcctttta aatggtagca atcaagtgaa ctatttggcc 120

tggtaccaac agaaaccagg acagtctcct aaacttctgg tgtactttgc gtccactagg 180

gaatctgggg tccctgatcg cttcataggc agtggatctg ggacagattt cactcttacc 240

atcagcagtg tgcaggctga agacctggca gattacttct gtcagcaaca ttatagcact 300

ccgtggacgt tcggtggagg caccaagctg gaaatcagac 340

<210>119

<211>322

<212>DNA

<213> light chain variable region coding sequence

<400>119

gacattgtga tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 60

atcacctgca aggccagtca ggatgttctt acatctgtag cctggtatca acagaaacca 120

gggcaatctc ctaaactcct gatttactcg gcatcctacc gatacactgg agtccctgat 180

cgcttcactg gcagtggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 240

gaagacctgg cagtttatta ctgtcagcaa cattatagta ctccgtggac gttcggtgga 300

ggctccaagc tggaaatcaa ac 322

<210>120

<211>354

<212>DNA

<213> heavy chain variable region coding sequence

<400>120

atgtgcagct tcaggagtcg ggacctggcc tggtgaaacc ttctcagtct ctgtccctca 60

cctgcactgt cactggctac tcaatcacca gtgattatgc ctggaactgg atccggcagt 120

ttccaggaaa caaactggag tggatgggct acataagcta cagtggtagc actaggtaca 180

acccatctct caaaagtcga atctctatca ctcgagacac atccaagaac cagttcttcc 240

tgcagttgaa ttctgtgact actgaggaca cagccacata ttactgtgca agaagggaac 300

tgggacaggg gtttccttac tggggccaag ggactctggt cactgtctct gcag 354

<210>121

<211>340

<212>PRT

<213> recombinant TIGIT extracellular region Fc fusion protein

<400>121

Met Met Thr Gly Thr Ile Glu Thr Thr Gly Asn Ile Ser Ala Glu Lys

1 5 10 15

Gly Gly Ser Ile Ile Leu Gln Cys His Leu Ser Ser Thr Thr Ala Gln

20 25 30

Val Thr Gln Val Asn Trp Glu Gln Gln Asp Gln Leu Leu Ala Ile Cys

35 40 45

Asn Ala Asp Leu Gly Trp His Ile Ser Pro Ser Phe Lys Asp Arg Val

50 55 60

Ala Pro Gly Pro Gly Leu GlyLeu Thr Leu Gln Ser Leu Thr Val Asn

65 70 75 80

Asp Thr Gly Glu Tyr Phe Cys Ile Tyr His Thr Tyr Pro Asp Gly Thr

85 90 95

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

100 105 110

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

115 120 125

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

130 135 140

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

145 150 155 160

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

165 170 175

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

180 185 190

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

195 200 205

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

210 215 220

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

225 230 235 240

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

245 250 255

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

260 265 270

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

275 280 285

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

290 295 300

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

305 310 315 320

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

325 330 335

Ser Pro Gly Lys

340

<210>122

<211>119

<212>PRT

<213> humanized 4A5 antibody heavy chain variable region sequence

<400>122

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

1 5 10 15

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

20 25 30

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

35 40 45

Met Gly Tyr Ile Thr Tyr Ser Gly Arg Thr Thr Tyr Asn Pro Ser Leu

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Trp Gly Leu Leu Arg Arg Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Leu Thr Val Ser Ser

115

<210>123

<211>107

<212>PRT

<213> humanized 4A5 antibody light chain variable region sequence

<400>123

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

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

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210>124

<211>327

<212>PRT

<213> human IgG4 heavy chain constant region sequence

<400>124

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

5055 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

210215 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>125

<211>107

<212>PRT

<213> human IgG4 kappa chain constant region sequence

<400>125

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>126

<211>446

<212>PRT

<213> heavy chain sequence of 4A5 chimeric antibody

<400>126

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

1 5 10 15

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

20 25 30

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

35 40 45

Met Gly Tyr Ile Thr Tyr Ser Gly Arg Thr Thr Tyr Asn Pro Ser Leu

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Trp Gly Leu Leu Arg Arg Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

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

115 120 125

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

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

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

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

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

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

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

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210>127

<211>214

<212>PRT

<213> light chain sequence of 4A5 chimeric antibody

<400>127

Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Phe Thr Ala

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala

65 70 75 80

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

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 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>128

<211>735

<212>DNA

<213> TIGIT protein coding sequence

<400>128

atgcgctggt gtctcctcct gatctgggcc caggggctga ggcaggctcc cctcgcctca 60

ggaatgatga caggcacaat agaaacaacg gggaacattt ctgcagagaa aggtggctct 120

atcatcttac aatgtcacct ctcctccacc acggcacaag tgacccaggt caactgggag 180

cagcaggacc agcttctggc catttgtaat gctgacttgg ggtggcacat ctccccatcc 240

ttcaaggatc gagtggcccc aggtcccggc ctgggcctca ccctccagtc gctgaccgtg 300

aacgatacag gggagtactt ctgcatctat cacacctacc ctgatgggac gtacactggg 360

agaatcttcc tggaggtcct agaaagctca gtggctgagc acggtgccag gttccagatt 420

ccattgcttg gagccatggc cgcgacgctg gtggtcatct gcacagcagt catcgtggtg 480

gtcgcgttga ctagaaagaa gaaagccctc agaatccatt ctgtggaagg tgacctcagg 540

agaaaatcag ctggacagga ggaatggagc cccagtgctc cctcaccccc aggaagctgt 600

gtccaggcag aagctgcacc tgctgggctc tgtggagagc agcggggaga ggactgtgcc 660

gagctgcatg actacttcaa tgtcctgagt tacagaagcc tgggtaactg cagcttcttc 720

acagagactg gttag 735

<210>129

<211>552

<212>PRT

<213> CD155-Fc protein sequence

<400>129

Trp Pro Pro Pro Gly Thr Gly Asp Val Val Val Gln Ala Pro Thr Gln

1 5 10 15

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

20 25 30

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

35 40 45

His Gly Glu Ser Gly Ser Met Ala Val Phe His Gln Thr Gln Gly Pro

50 55 60

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

65 70 75 80

Ala Glu Leu Arg Asn Ala Ser Leu Arg Met Phe Gly Leu Arg Val Glu

85 90 95

Asp Glu Gly Asn Tyr Thr Cys Leu Phe Val Thr Phe Pro Gln Gly Ser

100 105 110

Arg Ser Val Asp Ile Trp Leu Arg Val Leu Ala Lys Pro Gln Asn Thr

115 120 125

Ala Glu Val Gln Lys Val Gln Leu Thr Gly Glu Pro Val Pro Met Ala

130 135 140

Arg Cys Val Ser Thr Gly Gly Arg Pro Pro Ala Gln Ile Thr Trp His

145 150 155 160

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

165 170 175

Ser Gly Thr Val Thr Val Thr Ser Leu Trp Ile Leu Val Pro Ser Ser

180 185 190

Gln Val Asp Gly Lys Asn Val Thr Cys Lys Val Glu His Glu Ser Phe

195 200 205

Glu Lys Pro Gln Leu Leu Thr Val Asn Leu Thr Val Tyr Tyr Pro Pro

210 215 220

Glu Val Ser Ile Ser Gly Tyr Asp Asn Asn Trp Tyr Leu Gly Gln Asn

225 230 235 240

Glu Ala Thr Leu Thr Cys Asp Ala Arg Ser Asn Pro Glu Pro Thr Gly

245 250 255

Tyr Asn Trp Ser Thr Thr Met Gly Pro Leu Pro Pro Phe Ala Val Ala

260 265 270

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

275 280 285

Thr Leu Ile Cys Asn Val Thr Asn Ala Leu Gly Ala Arg Gln Ala Glu

290 295 300

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

305 310 315 320

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

325 330 335

Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

340 345 350

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

355 360 365

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

370 375 380

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

385 390 395 400

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

405 410 415

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

420 425 430

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

435 440 445

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

450 455 460

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

465 470 475 480

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

485 490 495

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

500 505 510

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

515 520 525

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

530 535 540

Ser Leu Ser Leu Ser Pro Gly Lys

545 550

Claims

1. An antibody capable of binding TIGIT, comprising a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2, and a light chain CDR 3;

wherein, the amino acid sequence of the heavy chain CDR1 is shown as SEQ ID NO. 20, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO. 21, and the amino acid sequence of the heavy chain CDR3 is shown as sequence table SEQ ID NO. 22; and the amino acid sequence of the light chain CDR1 is shown as SEQ ID NO. 6, the amino acid sequence of the light chain CDR2 is shown as SEQ ID NO. 7, and the amino acid sequence of the light chain CDR3 is shown as sequence table SEQ ID NO. 24.

2. The antibody of claim 1, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO. 19 and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO. 23.

3. The antibody of claim 1, wherein the antibody is capable of binding to the amino acid sequence set forth in SEQ ID NO. 121.

4. The antibody of any one of claims 1-3, wherein the antibody is a chimeric antibody or a humanized antibody.

5. An immunoconjugate comprising a therapeutic agent and the antibody of any one of claims 1-4 coupled to the therapeutic agent.

6. A composition comprising the antibody of any one of claims 1-4, and/or the immunoconjugate of claim 5, and a pharmaceutically acceptable carrier.

7. A kit for detecting TIGIT in a sample, the kit comprising the antibody of any one of claims 1-4.

8. Use of the antibody of any one of claims 1-4 in the preparation of a reagent for detecting TIGIT in a sample.

9. Use of an antibody according to any one of claims 1 to 4 in the manufacture of a medicament for the prevention and/or treatment of a CD155 expressing cancer.

10. The use of claim 9, wherein the cancer is at least one of lung cancer, liver cancer, ovarian cancer, cervical cancer, skin cancer, bladder cancer, colon cancer, breast cancer, glioma, kidney cancer, stomach cancer, esophageal cancer, oral squamous cell carcinoma, and head and neck cancer.

11. A nucleic acid encoding the antibody of any one of claims 1-4.

12. A recombinant vector or transformant containing the nucleic acid according to claim 11.