RU2786434C2 - Antibody to tigit and its use - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: present invention relates to biotechnology, in particular to a new antibody and a fragment of this antibody, which specifically bind to TIGIT, as well as to a composition including this antibody or its fragment. In addition, the present invention relates to nucleic acid encoding this antibody or its fragment, a host cell containing this nucleic acid, and their use. In addition, the present invention relates to therapeutic or diagnostic use of this antibody or its fragment.

EFFECT: obtaining an antibody to TIGIT.

21 cl, 9 dwg, 16 tbl, 6 ex

Description

FIELD OF TECHNOLOGY

The present invention relates to a new antibody and a fragment of this antibody that specifically binds to TIGIT, as well as to a composition comprising this antibody or its fragment. In addition, the present invention relates to a nucleic acid encoding this antibody or fragment thereof, a host cell containing this nucleic acid, and its use. In addition, the present invention relates to the therapeutic or diagnostic use of this antibody or its fragment.

BACKGROUND OF THE INVENTION

TIGIT (T-cell immune receptor comprising immunoglobulin (Ig) and ITIM domains, also known as WUCAM, Vstm3 or Vsig9) was first discovered as a member of the CD28 family in a bioinformatics comparison. TIGIT is a co-inhibitory receptor that is expressed on the surface membrane of various immune cells (natural killer (NK) cells, activated T lymphocytes, memory T cells, regulatory T cells, follicular T helpers, etc.) and is member of the immunoglobulin superfamily. It is believed that TIGIT molecules can regulate the immune system through three mechanisms: 1) TIGIT competes with the co-stimulatory CD226 receptor for binding to the common CD155/CD112 ligand expressed on the surface of dendritic or tumor cells and transmits inhibition signals to cells expressing TIGIT, thereby thereby suppressing the activation of these cells; 2) TIGIT interacts directly with the costimulatory CD226 receptor, preventing CD226 homodimerization by blocking its activation signal down the signal transduction chain; 3) after TIGIT binds to CD155, which is expressed by dendritic cells, the intracellular sequence of ITIM transmits inhibitory signals, resulting in the expression of immunosuppressive cytokines that suppress the activation of the immune system. TIGIT may play a role in a variety of cells that make up the tumor microenvironment. These cells may include tumor-infiltrating CD8 ⁺ T lymphocytes, regulatory T cells, and NK cells. In general, from a number of studies it follows that the immunosuppressive effect of TIGIT is mediated by the first of these mechanisms.

It has been shown that the binding of antibodies to the human TIGIT receptor can be used for the treatment of oncological diseases. See, for example, WO 2006/124667. Antibody blockade of PD-L1 and TIGIT can increase the CD8 ⁺ T cell-mediated tumor rejection in a synergistic manner in a mouse model. Grogan et al. (2014) J. Immunol. 192(1) Suppl. 203.15; Johnston et al. (2014) Cancer Cell 26:1-15. Similar results have been obtained in animal models of melanoma. Inozume et al. (2014) J. Invest. Dermatol. 134:S121-Abstract 693.

The role that TIGIT plays in the immune response makes it an attractive target for tumor immunotherapy. Accordingly, there is a need to develop new anti-TIGIT antibodies, in particular TIGIT-binding antibodies capable of blocking CD155/CD112, and more specifically, human anti-TIGIT antibodies, as well as combination preparations based on them, for the treatment of diseases, in particular for the treatment oncological diseases.

SUMMARY OF THE INVENTION

The subject of the present invention is an antibody to TIGIT, encoding this antibody gene, and their use. Using genetic engineering and yeast display techniques, the inventor screened a fully human anti-human TIGIT receptor antibody from a human antibody library present on the surface of yeast cells, and then obtained a high affinity anti-human TIGIT receptor antibody using affinity maturation. The fully humanized antibody molecule of the present invention is able to effectively block the binding of TIGIT to its CD155 ligand, attenuate or eliminate inhibition signals transmitted to cells, increase IL-2 production, and also suppress tumor growth when applied in vivo , and the effect of tumor suppression is particularly pronounced. when this antibody is used in combination with an anti-PD-1 antibody. Thus, the antibody of the present invention can be used for a variety of purposes, including, but not limited to, enhancing the immune response, suppressing tumor growth, combating infections, and detecting TIGIT proteins.

The subject of the present invention is a new fully humanized antibody that binds to the human TIGIT receptor and its antigen-binding fragment.

In some embodiments, an anti-TIGIT antibody of the present invention has one or more of the following properties:

(i) the ability to bind to human TIGIT with high affinity;

(ii) the ability to cross-react immunologically with monkey and/or mouse TIGIT;

(iii) efficient binding to TIGIT on the cell surface;

(iv) blocking the binding of TIGIT to its CD155 ligand;

(v) reducing the inhibitory effect of CD155 binding to TIGIT on IL-2 dependent downstream signaling;

(vi) increased production of IL-2 by T-lymphocytes;

(vii) antitumor activity, eg inhibition of tumor growth;

(viii) the ability to exert a more pronounced tumor inhibitory effect in combination with an anti-PD-1 antibody, eg enhanced tumor growth inhibition.

In some embodiments, the present invention provides an antibody or fragment thereof that binds to TIGIT, including: the sequences HCDR1, HCDR2, and HCDR3 of one of the heavy chain variable regions shown in SEQ ID NOs: 84-103, and/or light chain variable region LCDR1, LCDR2 and LCDR3 sequences set forth in SEQ ID NOs: 104-110, or a combination of the CDR sequences described above.

In some embodiments, the present invention also includes an anti-TIGIT antibody or antigen-binding fragment thereof. An anti-TIGIT antibody and an exemplary (type) antibody (e.g., an antibody combining a combination of the VH and VL antibody sequences listed in Table B) of the present invention bind to the same epitope or overlapping epitopes and/or compete for binding with TIGIT, and/or an anti-TIGIT antibody inhibits (eg, competitively inhibits) an exemplary antibody of the present invention.

In some embodiments of the invention, the subject of the invention is a nucleic acid encoding an antibody or antigen-binding fragment disclosed below, a vector containing the nucleic acid, and a host cell containing the vector.

In some embodiments of the invention, the subject of the present invention is a method for producing an antibody or antigen-binding fragment, disclosed below.

In some embodiments of the invention, the subject of the present invention is an immunoconjugate, a pharmaceutical composition and a combined preparation comprising an antibody of the present invention.

The subject of the present invention is also a method of blocking the binding of TIGIT to CD155 (eg CD155 expressed on the surface of a dendritic cell or tumor cell) in a subject using an antibody of the present invention. In some embodiments, TIGIT-mediated inhibition signals are attenuated or eliminated in cells (particularly T-lymphocytes and NK cells) expressing TIGIT in order to cause activation of T-lymphocytes and NK cells. In some embodiments of the invention due to the proposed method increases the production of IL-2 T-lymphocytes. In some embodiments, the method reduces CD155-mediated inhibition signaling in cells (eg, dendritic cells) expressing CD155, resulting in decreased expression of immunosuppressive cytokines. In some embodiments of the invention due to the proposed method is the activation of the immune system. Accordingly, the subject of the present invention is also a method for the prevention or treatment of neoplastic diseases or infection using the antibody of the present invention.

The present invention also relates to a method for detecting TIGIT in a sample.

The present invention is further illustrated by the following figures and specific examples. However, these drawings and specific examples of the invention should not be taken as limiting the scope of the invention, and the scope of protection defined by the claims will also include any modifications easily implemented by experts in the art and consistent with the essence of the present invention.

BRIEF DESCRIPTION OF THE FIGURES

RICE. 1A-F illustrate the binding of yeast-expressed antibodies to human TIGIT expressed on the surface of CHO cells before and after affinity maturation.

RICE. 2 illustrates the binding of 7 candidate antibodies expressed by CHO cells to human TIGIT expressed on the surface of CHO cells.

RICE. 3A-F illustrate the ability of yeast-expressed antibodies to block binding to CD155 before and after affinity maturation.

RICE. 4 illustrates the ability of 7 candidate antibodies expressed by CHO cells to block binding to CD155.

RICE. 5A-E illustrate the results of the analysis of the biological activity underlying the mechanism of action (MD), for antibodies expressed by yeast, before and after affinity maturation.

RICE. 6 illustrates the results of the analysis of the biological activity underlying MD for 7 candidate antibodies expressed by CHO cells.

RICE. 7 illustrates the results of pharmacodynamic studies of candidate molecules ADI-30293 and ADI-30278, which were administered separately (at a dose of 10 mg/kg) to MC38 mice with an inserted (knock-in) human TIGIT receptor gene.

RICE. 8A-B illustrate the results of pharmacodynamic studies of candidate molecules ADI-30293 and ADI-30278, which were administered (at a dose of 10+1 mg/kg) in combination with an anti-PD1 antibody (antibody C, WO 2017133540A1) to MC38 mice with an inserted (knock -in) human TIGIT receptor genome.

RICE. 9 illustrates an exemplary human TIGIT receptor sequence used in the examples of the present invention.

DETAILED DESCRIPTION

Definitions

Unless otherwise indicated, all technical and scientific terms used in this document have the conventional meaning given to them by those skilled in the art. For the purposes of the present invention, the following terms are defined below.

The term "about", used in combination with a numerical value, is intended to cover a range of numerical values from a lower limit that is 5% less than the specified numerical value to an upper limit that is 5% greater than the specified numerical value.

The term "and/or" should be understood to refer to any of the options, or to any two or more options.

As used in this document, the terms "comprise" or "include" should be understood to include elements, integers, or steps, but not as the exclusion of any other elements, integers, or steps. Used in this document, the terms "comprise" or "include", unless otherwise indicated, also refer to the situation where the whole consists of the described elements, integers or steps. For example, if it is said that an antibody variable region with a specific sequence is "included", it is also understood that an antibody variable region consisting of a specific sequence is included.

As used herein, the term "antibody" refers to a polypeptide comprising at least one immunoglobulin light chain variable region or immunoglobulin heavy chain variable region that specifically recognizes and binds to an antigen. The term "antibody" encompasses various antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, single or multi-chain antibodies, monospecific or polyspecific antibodies (e.g., bispecific antibodies), fully human or chimeric or humanized antibodies, as well as full length antibodies and fragments of antibodies, provided that they have the desired antigen-binding activity.

Those skilled in the art will appreciate that a "whole antibody" (in this document the term is used interchangeably with the terms "full antibody", "complete antibody", and "intact antibody") comprises at least two heavy chains (H) and two light chains (L). Each heavy chain consists of a heavy chain variable region (abbreviated as VH in this document) and a heavy chain constant region. The heavy chain constant region consists of 3 domains: CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated as VL in this document) and a light chain constant region. The light chain constant region consists of the CL domain. Variable regions are the heavy or light chain domains of antibodies that are involved in the binding of antibodies to their antigens. Constant regions are not directly involved in the binding of antibodies to antigens, but they perform a number of effector functions. Antibody light chains can be classified into one of two types, kappa (κ) and lambda (λ), depending on the amino acid sequence of their constant domain. Depending on the amino acid sequence of the heavy chain constant region, an antibody heavy chain can be classified into one of 5 major types, namely IgA, IgD, IgE, IgG, and IgM; some types of heavy chains can be further divided into subtypes, such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant regions corresponding to the 5 different types of antibodies are called α, δ, ε, γ, and μ, respectively. The term "isotype" refers to the type of antibody, which is determined by the constant region of the heavy chain of the antibody. See, for example, Fundamental Immunology , Ch.7 (Paul, w. Eds., 2nd ^edition , Raven Press, NY (1989)), which is incorporated herein by reference in its entirety and without reservation.

The term "antigen-binding portion" (in this document, the term is used interchangeably with the terms "antibody fragment" and "antigen-binding fragment") of an antibody refers to an incomplete antibody molecule that includes the part of the intact antibody for binding to the antigen to which the intact antibody binds. antibody. Those skilled in the art will appreciate that the antigen-binding portion of an antibody typically includes amino acid residues from a "complementarity determining region" or "CDR". The antigen-binding fragment can be obtained using recombinant DNA technology or by enzymatic or chemical cleavage of an intact antibody. Antigen binding fragments include, but are not limited to, Fab, scFab, Fab', F(ab') ₂ , Fab'-SH, Fv, single chain Fv, diabody, triatelo, tetrabody, minibody, and single domain antibody (sdAb). For more detailed descriptions of antibody fragments, see Fundamental Immunology , WE Paul eds., Raven Press, NY (1993); Shao Rongguang et al. (eds.), Antibody Drug Research and Application , People's Medical Publishing House (2013); Hollinger et al. , PNAS USA 90: 6444-6448 (1993); Hudson et al. , Nat. Med. 9:129-134 (2003).

The terms "human antibody" and "fully humanized antibody" are used interchangeably herein and refer to an antibody comprising variable regions in which the framework regions and CDR regions are derived from human immunoglobulin sequences in the germline configuration. In addition, if the antibody includes constant regions, then the constant regions are also derived from human immunoglobulin sequences in the germline configuration. A human antibody disclosed herein may include amino acid sequences (e.g., mutations introduced by in vitro random or site-directed mutagenesis or in vivo somatic mutagenesis) that are not present in germline-configured human immunoglobulin sequences, such as CDRs, and, in particular, in CDR3. However, the term "human antibody" as used herein does not include antibodies in which the CDR sequences are from the germline of other mammalian species (eg, mice) transplanted into human framework sequences.

As used herein, the term "recombinant human antibody" includes any human antibodies that are produced, expressed, produced, or isolated using recombinant DNA technology, such as: (a) antibodies isolated from transgenic or transchromosomal animals (e.g., mice) using human genes immunoglobulins or from hybridomas derived from human immunoglobulin genes; (b) antibodies isolated from host cells (eg, transfectome) transformed to express human antibodies; (c) antibodies isolated from recombinant or combinatorial human antibody libraries (eg yeast display libraries); (d) antibodies that are obtained, expressed, produced or isolated by any other means, including splicing of human immunoglobulin genes to obtain other DNA sequences. These recombinant human antibodies have variable regions in which the framework regions and CDR regions are derived from human immunoglobulin sequences in the germline configuration. However, in certain embodiments of the invention, recombinant human antibodies can be subjected to in vitro mutagenesis (or somatic in vivo mutagenesis in the case of transgenic animals using a human Ig sequence), and the amino acid sequences of the VH and VL regions of the recombinant antibodies thus obtained, although they are originate from human germline VH and VL sequences, do not occur in a human antibody library based on natural germline sequences.

The term "chimeric antibody" refers to an antibody in which the variable region sequences are from one species and the constant region sequences are from another species, such as an antibody in which the variable region sequences are from a mouse antibody and the constant region sequences are from a human antibody.

The term "humanized antibody" refers to an antibody in which CDR sequences derived from a mammal of another species, such as a mouse, are fused to human framework sequences. Additional modifications may be made to human framework sequences.

An "isolated" antibody is an antibody that has been separated from components of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, as determined, for example, by electrophoresis (e.g., DNS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for determining antibody purity, see, for example, Flatman, S. et al. , J.Chrom.B 848 (2007) 79-87.

The term "epitope" refers to the region of an antigen to which an antibody binds. Epitopes can be formed by both amino acids constituting a continuous sequence, and amino acids that do not constitute a continuous sequence, but are brought together in space when the protein is folded into a tertiary structure.

In this document, TIGIT refers to "T-cell immune receptor comprising the Ig and ITIM domains". The term also includes variants, isotypes, homologues, and species homologues of TIGIT. The term “human TIGIT” refers to a human TIGIT sequence. One specific sequence of human TIGIT is shown in SEQ ID NO: 208. In some embodiments, the amino acid sequence of human TIGIT is at least 95%, and even at least 96%, 97%, 98%, or 99% identical to the amino acid sequence Human TIGIT as set forth in SEQ ID NO: 208. TIGIT proteins may also include fragments of TIGIT, such as fragments containing extracellular domains, such as fragments retaining the ability to bind to any of the antibodies disclosed herein. In this document, CD155, also known as PVR (poliovirus receptor), PVS, HVED, CD155, NECL5, TAGE4 and Necl-5, interacts with TIGIT to induce immunosuppression signals.

The term "specifically binds to" means that the antibody selectively or preferentially binds to the antigen. If the antibody binds to human TIGIT with a K _D of about 5×10 ^-7 M or less, about 1×10 ^-7 M or less, about 5×10 ^-8 M or less, about 1×10 ^-8 M or less, about 5×10 ^-9 M or less, as determined by optical interferometry, then this antibody is an antibody that "specifically binds to human TIGIT". However, an antibody that specifically binds to human TIGIT may be cross-reactive with TIGIT proteins from other species. For example, in some embodiments, an antibody that specifically binds to human TIGIT may cross-react with TIGIT proteins from non-human species. In other embodiments, an antibody that specifically binds to human TIGIT may be completely specific for human TIGIT and not cross-reactive with other species, or only cross-reactive with TIGIT from certain species.

As used herein, the term "cross-reactivity" refers to the ability of an antibody to bind to TIGIT from different species. For example, an antibody described herein that binds to human TIGIT may also bind to TIGIT from other species (eg, monkey and/or mouse TIGIT). The method for determining cross-reactivity includes the method described in the examples, as well as standard methods of analysis known to those skilled in the art, such as biological optical interferometry or flow cytometry.

The terms "affinity" or "binding affinity" refer to the binding affinity inherent in a given pair of molecules, which reflects the characteristics of the interaction between two molecules that bind to each other. The affinity of the X molecule for partner Y can generally be characterized by the equilibrium dissociation constant (K _D ), which is the ratio of the dissociation rate constant (k _dis ) and the association rate constant (k _on ). Affinity can be measured by conventional methods known to those skilled in the art. One method for measuring affinity is the ForteBio binding kinetic assay described in this document.

In the case of an IgG antibody, the term "high affinity" means that the antibody binds to the target antigen with a K _D of 1×10 ^-7 M or less, preferably 5×10 ^-8 M or less, more preferably about 1×10 ^{- 8} M or less, most preferably about 5×10 ^-9 M or less. However, "high affinity" binding can be understood differently depending on the isotype of the antibody. For example, for an IgM isotype antibody, "high affinity" means that the antibody has a K _D of 1×10 ^-6 M or less, preferably 1×10 ^-7 M or less, more preferably about 1×10 ^-8 M or less.

The term "antibody competing for binding" to an antigen (eg, TIGIT) with a reference antibody means that the antibody blocks binding of the reference antibody to the antigen (eg, TIGIT) by 50% or more in a competitive binding assay and, conversely, the reference antibody blocks binding of a given antibody with an antigen (eg, TIGIT) by 50% or more in a competitive binding assay. Examples of competitive binding assays are described in "Antibodies", Harbor and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY). An antibody competing for binding and a reference antibody can bind to the same epitope region, eg, the same epitope, adjacent epitopes, or overlapping epitopes.

An antibody that inhibits (e.g., competitively inhibits) the binding of a reference antibody to its antigen is understood to mean an antibody that inhibits the binding of a reference antibody to its antigen by 50%, 60%, 70%, 80%, 90%, 95% or more. Conversely, a reference antibody inhibits the binding of that antibody to its antigen by 50%, 60%, 70%, 80%, 90%, 95% or more. The binding of an antibody to an antigen can be characterized by affinity (eg, equilibrium dissociation constant). Methods for determining affinity are known to those skilled in the art.

By an antibody exhibiting the same or similar binding affinity and/or specificity as a reference antibody is meant an antibody capable of exhibiting a binding affinity and/or specificity of at least 50%, 60%, 70%, 80%, 90%, 95% or more of those of the reference antibody. This can be determined by any method used to determine binding affinity and/or specificity and known to those skilled in the art.

The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain, including at least a portion of the constant region. The term includes a native sequence Fc region and a variant Fc region. In one embodiment, the Fc region of a human immunoglobulin heavy chain starts at a Cys226 or Pro230 residue of the heavy chain and ends at the carboxyl terminus. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise indicated, throughout this document, the numbering of amino acid residues in an Fc region or constant region is based on the EU numbering system, also known as the EU index, as described in Kabat, EA, et al. , Sequences of Proteins of Immunological Interest , 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD (1991), NIH Publication ^91-3242 .

The term "variant" in relation to an antibody, as used herein, refers to an antibody comprising a target region of the antibody in which the amino acids are changed from the reference antibody by at least one modification, e.g., 1-30, 1-20, or 1-10 modifications. , for example, by substitutions, deletions, and/or insertions of 1, 2, 3, 4, or 5 amino acids, and wherein the variant substantially retains at least one biological property (e.g., the ability to bind to an antigen) of the antibody molecule prior to the change. The target region of an antibody can be a full-length antibody, or a heavy chain variable region, or a light chain variable region, or a combination thereof, or one or more heavy chain CDRs, or one or more light chain CDRs, or a combination thereof. Herein, a region of an antibody in which the amino acids are changed from that of the reference antibody is also referred to as a "variant" region of the reference antibody.

As used herein, the term "sequence identity" refers to the degree of nucleotide or amino acid sequence identity within a comparison window. "Percent sequence identity" can be calculated using the following steps: comparing two optimally matched sequences in a comparison window; determination of the number of positions in which identical nucleotide residues ( e.g. , A, T, C, G, and I) or amino acids ( e.g. , Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp , Lys, Arg, His, Asp, Glu, Asn, Gln, Cys, and Met) occur in both sequences, resulting in the number of corresponding (correctly paired) positions; dividing the number of correctly paired positions by the total number of positions in the comparison window ( i.e., window size); and multiplying the result by 100 to obtain percent sequence identity. Optimal alignment for determining percent sequence identity can be achieved by various methods known to those skilled in the art, for example using publicly available computer programs such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR). Those of skill in the art can determine the appropriate sequence matching parameters, including the algorithm needed to optimally match compared sequences, whether full length or targeted.

As applied to antibody sequences, the percent amino acid sequence identity herein is determined by optimal matching of the candidate antibody sequence with the reference antibody sequence, and, in a preferred embodiment of the invention, the optimal matching is performed according to the Kabat numbering scheme. Herein, if no comparison window (ie, the target region of the antibody to be compared) is specified, then the matching is performed against the full sequence of the reference antibody. In some embodiments of the invention, as far as antibodies are concerned, sequence identity may be provided for the heavy chain variable region and/or light chain variable region, or the percent sequence identity may refer only to the framework regions, while the sequences of the corresponding CDRs plots remain 100% identical.

Similarly, with respect to antibody sequences, a candidate antibody with a target region in which the amino acids are changed compared to the reference antibody can be determined based on the alignment.

As used herein, the term "conservative substitution" refers to an amino acid change that replaces an amino acid with a chemically similar amino acid. Amino acid modifications, such as substitutions, can be introduced into the antibody of the present invention by standard methods known to those skilled in the art, such as site-directed mutagenesis or PCR-mediated mutagenesis.

Conservative substitution tables that present functionally similar amino acids are well known to those skilled in the art. In a preferred aspect, conservatively substituted residues are selected from Table A of conservative substitutions below, and preferably are the preferred conservatively substituted residues shown in Table A.

Table A

Elementary remainder Replacement example Preferred conservative substitution Ala (A) Val; Leu; ile Val Arg(R) Lys; gln; Asn Lys Asn(N) gln; His; asp; Lys; Arg Gln Asp(D) Glu; Asn Glu Cys(C) Ser; Ala Ser Gln(Q) asn; Glu Asn Glu(E) asp; Gln asp Gly (G) Ala Ala His(H) asn; gln; Lys; Arg Arg Ile (I) Leu; Val; met; Ala; Ph; Nle Leu Leu(L) Nle; Ile; Val; met; Ala; Phe ile Lys (K) Arg; gln; Asn Arg Met(M) Leu; Ph; ile Leu Phe(F) trp; Leu; Val; Ile; Ala; Tyr Tyr Pro (P) Ala Ala Ser(S) Thr Thr Thr(T) Val; Ser Ser TRP(W) Tyr; Phe Tyr Tyr (Y) trp; Ph; Thr; Ser Phe Val(V) Ile; Leu; met; Ph; Ala; Nle Leu

All aspects of the present invention are discussed in detail in the following sections.

I. TIGIT Antibody of the Invention

One aspect of the present invention is an antibody or antigen-binding fragment thereof, in particular a fully humanized antibody or antigen-binding fragment thereof, that specifically binds to TIGIT, preferably a human TIGIT protein ( e.g. , the human TIGIT sequence shown in SEQ ID NO : 208). In some embodiments, the antigen-binding fragment of an antibody disclosed herein is an antibody fragment selected from the following list: Fab, Fab', Fab'-SH, Fv, single chain antibodies such as scFv, (Fab')2 α-fragments, single domain antibodies, diabodies (dAbs), or linear antibodies.

Biological properties of antibodies that provide their benefits

In some embodiments, an anti-TIGIT antibody or antigen-binding fragment thereof disclosed herein binds to human TIGIT with high affinity, such as an equilibrium dissociation constant (K _D ) of less than 100×10 ^-9 M, less than or equal to about 50 ×10 ^-9 M, preferably less than or equal to about 1-30x10 ^-9 M, and more preferably about ^{1x10 -9} M, even more preferably about 1-10x10 ^-10 M. in a preferred embodiment, K _D is determined by biological optical interferometry (eg, by Fortebio affinity analysis). In some embodiments, K _D is determined by measuring the monovalent binding affinity of a Fab fragment (eg, a yeast expressed Fab fragment) of an antibody to human TIGIT. In a preferred embodiment, K _D for monovalent interaction is 1-100×10 ^-10 M, more preferably 1-50×10 ^-10 M, even more preferably 1-10×10 ^-10 M or 2-5×10 ^-10 M. In other embodiments, K _D is determined by measuring the monovalent binding affinity of an intact antibody (eg, an intact antibody expressed by CHO cells) to human TIGIT. In a preferred embodiment, K _D for monovalent interaction is 1-50×10 ^-10 M, more preferably 1-30×10 ^-10 M or 1-10×10 ^-10 M.

In some embodiments, an anti-TIGIT antibody or antigen-binding fragment thereof disclosed herein is cross-reactive with the monkey TIGIT protein. In some embodiments, the antibody binds to monkey TIGIT with high affinity, wherein the K _D (e.g., determined by measuring the monovalent binding affinity of an intact antibody to monkey TIGIT) is about 0.1-100×10 ^-9 M, more preferably 0 , 1-50×10 ^-9 M or 1-30×10 ^-10 M. In some embodiments, the antibody has cross-reactivity with the mouse TIGIT protein, with K _D (for example, determined by measuring the monovalent binding affinity of an intact antibody to mouse TIGIT ) is about 1-100×10 ^-9 M, such as 1-10×10 ^-7 M or 1-10×10 ^-8 M, or 1-10×10 ^-9 M. In other embodiments, the anti-TIGIT antibody of of the present invention does not cross-react with mouse TIGIT.

In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein binds to TIGIT expressed on the surface of cells with high affinity. In one embodiment, the cells expressing human TIGIT on their surface are CHO cells. In a preferred embodiment, the EC ₅₀ value, indicative of antibody binding to cells expressing human TIGIT, is measured by flow cytometry (eg, FACS). In some embodiments, the antibody is an intact yeast-expressed antibody with an EC _{50 of} less than about 10 nM, such as 0.1-1 nM, preferably less than or equal to about 1 nM, and more preferably about 0.2- 0.9 nM, such as 0.9 nM, 0.6 nM, or 0.4 nM. In some embodiments, the antibody is an intact CHO cell-expressed antibody with an EC _{50 of} less than about 10 nM, such as 0.1-1 nM, preferably about 0.1-0.3 nM, such as about 0.3 nM, 0.2 nM, or 0.1 nM.

In some embodiments, the antibody or antigen-binding fragment disclosed herein inhibits the corresponding activity of TIGIT. In some embodiments of this invention, the antibody or antigen-binding fragment disclosed herein blocks the binding of TIGIT to its CD155 ligand. In a preferred embodiment, the ability of an antibody to block the binding of human TIGIT (TIGIT expressed on cells) to human CD155 (eg, IC ₅₀ ) is measured by flow cytometry (eg, FACS). In some embodiments, the antibody is an intact yeast-expressed antibody with an IC _{50 of} less than about 10 nM, such as 0.1-2 nM, preferably about 0.1-1.0 nM, such as 0.8 nM, 0.6 nM, 0.4 nM or 0.2 nM. In other embodiments, the antibody is an intact antibody expressed by CHO cells with an EC _{50 of} less than about 1 nM, such as 0.1-0.5 nM, preferably about 0.3 nM, 0.2 nM, or 0.1 nM.

In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein attenuates or eliminates the inhibition signals caused by TIGIT binding to CD155. In some embodiments, an antibody or antigen-binding fragment thereof disclosed herein attenuates or eliminates TIGIT-mediated inhibition signaling in a cell (particularly a T-lymphocyte) expressing TIGIT. In some embodiments, an antibody or antigen-binding fragment thereof disclosed herein induces T-lymphocyte expression of genes downstream of IL-2 promoters, and in some embodiments, increases IL-2 production in T-lymphocytes. In some embodiments, the ability of an antibody to attenuate or eliminate inhibitory signals induced by TIGIT binding to CD155 (e.g., EC ₅₀ ) is determined by a fluorescent reporter protein assay (e.g., the MD-based assay of Example 5). In some embodiments, the antibody is an intact yeast-expressed antibody with an EC ₅₀ preferably less than about 10 nM, such as 0.1-5 nM, and more preferably about 0.1-3.0 nM, such as about 2.0 nM, 1.5 nM, 1.0 nM or 0.5 nM. In other embodiments, the antibody is an intact CHO cell-expressed antibody with an EC ₅₀ preferably less than 5 nM, such as about 0.1-3.0 nM, such as about 1.6 nM, 1.2 nM, 1 .0 nM or about 0.18-0.5 nM.

In some embodiments, an antibody or antigen-binding fragment thereof disclosed herein inhibits the growth of a tumor including infiltrating lymphocytes that express human TIGIT. In one embodiment, the tumor is a tumor of the gastrointestinal tract, preferably colorectal cancer. For example, in an in vivo transplanted tumor model, for example in MC38 mice, the growth of colon cancer is inhibited. In some embodiments, combination therapy with an antibody of the present invention and an anti-PD-1 antibody provides a much greater antitumor effect than either antibody alone.

In a preferred embodiment, an antibody or antigen-binding fragment disclosed herein has at least one, preferably at least two, more preferably at least three, four or five, and even more preferably all of the above. properties.

CDR regions of antibodies

"complementarity determining region", "CDR region" or "CDR" (these terms are used interchangeably with the term "hypervariable region" or "HVR" in this document) is an amino acid-derived region in the variable region of an antibody that is primarily responsible for binding to an antigen epitope. The heavy and light chain CDRs are generally referred to as CDR1, CDR2, and CDR3, with the numbering increasing from the N-terminus. The CDRs located in the heavy chain variable region of an antibody are referred to as HCDR1, HCDR2 and HCDR3, while the CDRs located in the antibody light chain variable region are referred to as LCDR1, LCDR2 and LCDR3.

VH and VL sequence combinations of some exemplary antibodies disclosed herein are listed in Table B below:

Antibody VH comprising or consisting of the amino acid sequence of the following SEQ ID NO VL comprising or consisting of the amino acid sequence with the following SEQ ID NO one SEQ ID NO:84 SEQ ID NO:104 2 SEQ ID NO:85 SEQ ID NO:104 3 SEQ ID NO:86 SEQ ID NO:104 4 SEQ ID NO:87 SEQ ID NO:104 five SEQ ID NO:88 SEQ ID NO:105 6 SEQ ID NO:89 SEQ ID NO:105 7 SEQ ID NO:90 SEQ ID NO:106 eight SEQ ID NO:91 SEQ ID NO:107 nine SEQ ID NO:92 SEQ ID NO:107 10 SEQ ID NO:93 SEQ ID NO:107 eleven SEQ ID NO:94 SEQ ID NO:108 12 SEQ ID NO:95 SEQ ID NO:108 13 SEQ ID NO:96 SEQ ID NO:108 fourteen SEQ ID NO:97 SEQ ID NO:108 fifteen SEQ ID NO:98 SEQ ID NO:109 16 SEQ ID NO:99 SEQ ID NO:109 17 SEQ ID NO:100 SEQ ID NO:109 eighteen SEQ ID NO:101 SEQ ID NO:110 nineteen SEQ ID NO:102 SEQ ID NO:110 twenty SEQ ID NO:103 SEQ ID NO:110

Those skilled in the art are aware of various schemes (methods) for identifying a CDR sequence for a given VH or VL amino acid sequence. For example, the Kabat method based on sequence variability is most commonly used to identify complementarity determining regions (CDRs) (Kabat et al. , Sequences of Proteins of Immunological Interest , 5th ^Ed . Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The Chothia method is based on the location of structural loops (Chothia and Lesk, J. mol. biol. 196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software. The definition of "contact" HVR-regions is based on the analysis of the existing complex crystal structures. According to various CDR definition schemes, the residues included in each HVR/CDR for these HVR regions are defined as follows.

CDR Kabat method AbM method Chothia Method Contact method LCDR1 L24-L34 L24-L34 L26-L32 L30-L36 LCDR2 L50-L56 L50-L56 L50-L52 L46-L55 LCDR3 L89-L97 L89-L97 L91-L96 L89-L96 HCDR1 H31-H35B H26-H35B H26-H32 H30-H35B (Kabat numbering system) HCDR1 H31-H35 H26-H35 H26-H32 H30-H35 (Chothia numbering system) HCDR2 H50-H65 H50-H58 H53-H55 H47-H58 HDDR3 H95-H102 H95-H102 H96-H101 H93-H101 (Kabat numbering system)

HVR regions can also be HVR sequences defined by the following residue positions according to the Kabat numbering system:

positions 24-36 or 24-34 (LCDR1), positions 46-56 or 50-56 (LCDR2), and positions 89-97 or 89-96 (LCDR3) in VL; regulations 26-35 or 27-35B (HCDR1), regulations 50-65 or 49-65 (HCDR2), and regulations 93-102, 94-102 or 95-102 (HCDR3) in VH.

In one embodiment, the HVR regions of an antibody disclosed herein are HVR sequences defined by the following residue positions according to the Kabat numbering system:

positions 24-34 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3) in VL, and positions 27-35B (HCDR1), 50-65 (HCDR2) and 93-102 (HCDR3) in VH.

In one embodiment, the HVR regions of an antibody disclosed herein are HVR sequences defined by the following residue positions according to the Kabat numbering system:

positions 24-34 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3) in VL, and positions 26-35B (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3) in VH.

HVR regions can also be defined based on residue positions according to the Kabat numbering system corresponding to a CDR reference sequence (eg, any of the exemplary CDRs disclosed herein).

Unless otherwise indicated, the terms "CDR", "CDR sequence", "HVR", or "HVR sequence" as used herein include CDR or HVR sequences defined by any of the methods described above.

Unless otherwise indicated, herein, positions of antibody variable region residues (including heavy chain variable region residues and light chain variable region residues) in this invention are numbered according to the Kabat numbering system (Kabat et al. , Sequences of Proteins of Immunological Interest , 5th ^Ed , Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

In one preferred embodiment of the invention, the CDR sequences disclosed herein are shown in Table 2, where HCDR1 is the CDR sequence determined by the AbM method and the remaining CDRs are the CDR sequences determined by the Kabat method.

In another preferred embodiment of the invention, the CDR sequences disclosed herein are shown in Table 1.

Combinations of some illustrative CDR sequences disclosed herein are listed in Table C below:

Combinations HCDR1 comprising or consisting of the amino acid sequence of the following SEQ ID NO HCDR2 comprising or consisting of the amino acid sequence of the following SEQ ID NO HCDR3 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR1 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR2 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR3 comprising or consisting of the amino acid sequence of the following SEQ ID NO one SEQID NO:1 SEQ ID NO:6 SEQ ID NO:11 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 2 SEQ ID NO:2 SEQ ID NO:7 SEQ ID NO:12 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 3 SEQ ID NO:3 SEQ ID NO:8 SEQ ID NO:13 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 4 SEQ ID NO:4 SEQ ID NO:9 SEQ ID NO:14 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 five SEQ ID NO:5 SEQ ID NO:10 SEQ ID NO:15 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 6 SEQ ID NO:19 SEQ ID NO:22 SEQ ID NO:25 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:28 7 SEQ ID NO:20 SEQ ID NO:23 SEQ ID NO:25 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:28 eight SEQ ID NO:19 SEQ ID NO:22 SEQ ID NO:25 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:29 nine SEQ ID NO:21 SEQ ID NO:24 SEQ ID NO:25 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:30 10 SEQ ID NO:31 SEQ ID NO:34 SEQ ID NO:38 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 eleven SEQ ID NO:31 SEQ ID NO:35 SEQ ID NO:38 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 12 SEQ ID NO:32 SEQ ID NO:36 SEQ ID NO:39 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 13 SEQ ID NO:33 SEQ ID NO:37 SEQ ID NO:40 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 fourteen SEQ ID NO:44 SEQ ID NO:48 SEQ ID NO:53 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 fifteen SEQ ID NO:45 SEQ ID NO:49 SEQ ID NO:53 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 16 SEQ ID NO:46 SEQ ID NO:50 SEQ ID NO:54 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 17 SEQ ID NO:46 SEQ ID NO:51 SEQ ID NO:55 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 eighteen SEQ ID NO:47 SEQ ID NO:52 SEQ ID NO:56 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 nineteen SEQ ID NO:60 SEQ ID NO:6 SEQ ID NO:66 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 twenty SEQ ID NO:61 SEQ ID NO:63 SEQ ID NO:66 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 21 SEQ ID NO:61 SEQ ID NO:64 SEQ ID NO:67 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 22 SEQ ID NO:62 SEQ ID NO:65 SEQ ID NO:68 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 23 SEQ ID NO:71 SEQ ID NO:22 SEQ ID NO:78 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83 24 SEQ ID NO:72 SEQ ID NO:75 SEQ ID NO:78 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83 25 SEQ ID NO:73 SEQ ID NO:76 SEQ ID NO:79 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83 26 SEQ ID NO:74 SEQ ID NO:77 SEQ ID NO:80 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83

Combinations of some illustrative CDR sequences disclosed herein are listed in Table D below:

Combinations HCDR1 comprising or consisting of the amino acid sequence of the following SEQ ID NO HCDR2 comprising or consisting of the amino acid sequence of the following SEQ ID NO HCDR3 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR1 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR2 comprising or consisting of the amino acid sequence of the following SEQ ID NO An LCDR3 comprising or consisting of the amino acid sequence of the following SEQ ID NO one SEQ ID NO:178 SEQ ID NO:6 SEQ ID NO:182 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 2 SEQ ID NO:179 SEQ ID NO:7 SEQ ID NO:183 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 3 SEQ ID NO:180 SEQ ID NO:8 SEQ ID NO:184 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 4 SEQ ID NO:181 SEQ ID NO:9 SEQ ID NO:185 SEQ ID NO:16 SEQ ID NO:17 SEQ ID NO:18 five SEQ ID NO:186 SEQ ID NO:22 SEQ ID NO:188 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:28 6 SEQ ID NO:187 SEQ ID NO:23 SEQ ID NO:188 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:28 7 SEQ ID NO:186 SEQ ID NO:22 SEQ ID NO:188 SEQ ID NO:26 SEQ ID NO:27 SEQ ID NO:29 eight SEQ ID NO:189 SEQ ID NO:34 SEQ ID NO:191 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 nine SEQ ID NO:189 SEQ ID NO:35 SEQ ID NO:191 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 10 SEQ ID NO:190 SEQ ID NO:36 SEQ ID NO:192 SEQ ID NO:41 SEQ ID NO:42 SEQ ID NO:43 eleven SEQ ID NO:193 SEQ ID NO:48 SEQ ID NO:196 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 12 SEQ ID NO:194 SEQ ID NO:49 SEQ ID NO:196 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 13 SEQ ID NO:195 SEQ ID NO:50 SEQ ID NO:197 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 fourteen SEQ ID NO:195 SEQ ID NO:51 SEQ ID NO:198 SEQ ID NO:57 SEQ ID NO:58 SEQ ID NO:59 fifteen SEQ ID NO:199 SEQ ID NO:6 SEQ ID NO:201 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 16 SEQ ID NO:200 SEQ ID NO:63 SEQ ID NO:201 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 17 SEQ ID NO:200 SEQ ID NO:64 SEQ ID NO:202 SEQ ID NO:69 SEQ ID NO:17 SEQ ID NO:70 eighteen SEQ ID NO:203 SEQ ID NO:22 SEQ ID NO:206 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83 nineteen SEQ ID NO:204 SEQ ID NO:75 SEQ ID NO:206 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83 twenty SEQ ID NO:205 SEQ ID NO:76 SEQ ID NO:207 SEQ ID NO:81 SEQ ID NO:82 SEQ ID NO:83

Antibodies that differ in specificity (i.e., have different binding sites for different antigens) also differ in their CDRs. However, although different antibodies differ in their CDRs, only a limited number of amino acid residues within the CDR are directly involved in antigen binding. Using at least two of the methods described (Kabat, Chothia, AbM, and the contact method), the smallest overlapping region can be determined, which allows a "minimum binding unit" for antigen binding to be determined. The minimum linking unit may be a CDR subfragment. Those skilled in the art will recognize that residues included in the remaining CDR sequences can be determined based on knowledge of antibody structure and protein folding. Therefore, any CDR options given in this document are also considered in this invention. For example, in one CDR embodiment, the amino acid residues in the minimum binding unit may remain unchanged, while other CDR residues defined by Kabat or Chothia may be replaced by conservative amino acid residues.

In some embodiments, an antibody of the present invention contains at least one, two, three, four, five, or six CDRs that are identical to, or are variants of, the corresponding CDRs within the variable region sequences of any of the antibodies listed in Table B. In some embodiments, an antibody of the present invention contains at least one, two, or three HCDRs that are identical to, or are variants of, the corresponding heavy chain CDRs within the variable region sequences of any of the antibodies listed in Table B. In some embodiments, the antibody of the present invention of the invention contains at least one, two, or three LCDRs that are identical to, or are variants of, corresponding to the light chain CDRs within the variable region sequences of any of the antibodies listed in Table B. As used herein, the term "corresponding CDRs" refers to CDRs that are located are located within the amino acid sequences of the variable region of the candidate antibody in such a way that their location most closely resembles that of the CDRs in the reference antibody after optimal matching. As used herein, a CDR variant is a CDR that has been modified by substitution, deletion, and/or insertion of at least one amino acid, such as one or two or three amino acids, and wherein the antigen-binding molecule containing the CDR variant is substantially retains the biological properties of the antigen-binding molecule containing the unmodified CDRs, eg retains at least 60%, 70%, 80%, 90%, or 100% biological activity (eg, antigen-binding capacity). It should be understood that each CDR may be modified independently or in combination. In a preferred embodiment, the amino acid modification is an amino acid substitution, in particular a conservative amino acid substitution, such as a substitution of one of the conservative amino acids listed in Table A. In some embodiments, amino acid substitutions are preferably made at amino acid positions corresponding to residues X of the CDR consensus sequences (e.g. shown in SEQ ID NOs: 5, 10,15, 21, 24, 30, 33, 37, 40, 47, 52, 56, 62, 65, 68, 74, 77 and 80) listed in this document.

In addition, those skilled in the art will recognize that a CDR3 region that is independent of CDR1 and/or CDR2 regions may itself determine the binding specificity of an antibody to its antigen. Also, many other antibodies with the same binding specificity can be generated based on the CDR3 consensus sequence. See, for example, US Pat. Nos. 6,951,646; 6,914,128; 6,090,382; 6,818,216; 6,156,313; 6,827,925; 5,833,943; 5,762,905 and 5,760,185. All of these patents are incorporated herein by reference.

Thus, in one embodiment, an antibody of the present invention comprises a CDR3 sequence from the heavy and/or light chain variable region of any of the antibodies listed in Table B, wherein the antibody is capable of specifically binding to human TIGIT. In yet another embodiment, the antibody may further comprise a CDR2 from the heavy and/or light chain variable region of the same antibody, or a CDR2 from the heavy and/or light chain variable region of another anti-TIGIT antibody. In yet another embodiment, the antibody may further comprise a CDR1 from the heavy and/or light chain variable region of the same antibody, or a CDR1 from the heavy and/or light chain variable region of another anti-TIGIT antibody. The activity of these antibodies, including the ability to bind to human TIGIT, the ability to block the binding of TIGIT to CD155 molecules, and/or the ability to suppress tumor growth, can be characterized by the analytical methods described herein.

In another aspect, given that antigen-binding specificity depends primarily on the CDR1, CDR2, and CDR3 regions, in some embodiments, the VH CDR sequences 1, 2, and 3, and the VL CDR sequences 1, 2, and 3 can be “combined and combined paired" (e.g., CDRs from different antibodies that bind to the same TIGIT antigen can be combined and paired, and each antibody preferably includes VH CDRs 1, 2, and 3, and VL CDRs 1, 2, and 3) to obtain other molecules of the present invention that bind to TIGIT. The binding of such "combined and paired" antibodies to TIGIT can be studied by binding assays known to those skilled in the art (eg, ELISA, SET, and Biacore) as well as other assays described in the examples. When VH CDR sequences are combined and paired, the CDR1, CDR2 and/or CDR3 sequences from a particular VH sequence are preferably replaced by structurally similar CDR sequences. Similarly, when VL CDR sequences are combined and paired, the CDR1, CDR2 and/or CDR3 sequences from a particular VL sequence are preferably replaced with structurally similar CDR sequences. The CDRs can be "combined and paired" for the antibodies of the present invention shown in Table 3. In addition, those skilled in the art will appreciate that the antibodies of the present invention may also be generated by substituting the structurally similar CDR sequences disclosed in this document to one or more VH CDR and/or VL CDR sequences from other different antibodies.

Thus, in some embodiments, an antibody or antigen-binding fragment thereof disclosed herein comprises a heavy chain variable region comprising a heavy chain complementarity determining region 3 (HCDR3), wherein HCDR3:

(i) identical to the heavy chain variable region HCDR3 of any of the antibodies listed in Table B;

(ii) identical to any of the HCDR3 sequences listed in Table C or D; or

(iii) includes at least 1 (preferably 1-2, and more preferably 1) amino acid modification (preferably a substitution, and more preferably a conservative substitution) compared to HCDR3 in item (i) or item (ii).

In some embodiments, an antibody or antigen-binding fragment disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the complementarity determining regions 3 of the heavy chain (HCDR3) and the complementarity determining regions 3 of the light chain (LCDR3) of the antibody :

(i) identical to the heavy and light chain variable region HCDR3 and LCDR3 sequences of any of the antibodies listed in Table B;

(ii) identical to the HCDR3 and LCDR3 sequences listed in Table C or D, in any combination; or

(iii) include a total of at least 1 (preferably 1-2, and more preferably 1) amino acid modification (preferably a substitution, and more preferably a conservative substitution) compared to HCDR3 and LCDR3 in item (i) or item (ii).

In one embodiment, an antibody or antigen-binding fragment thereof disclosed herein comprises a heavy chain variable region (VH), wherein the VH includes:

(i) the HCDR1, HCDR2 and HCDR3 sequences contained in the VH sequence of any of the antibodies listed in Table B;

(ii) the HCDR1, HCDR2 and HCDR3 sequences listed in Table C or D, in any combination; or

(iii) sequences having a total of at least one and no more than 5, 4, 3, 2 or 1 amino acid modifications (preferably amino acid substitutions, preferably conservative substitutions) in three CDRs compared to the sequences of point (i) or point (ii) ).

In another embodiment, an antibody or antigen-binding fragment thereof disclosed herein comprises a light chain variable region (VL), wherein VL includes:

(i) the LCDR1, LCDR2 and LCDR3 sequences contained in the VL sequence of any of the antibodies listed in Table B;

(ii) the LCDR1, LCDR2 and LCDR3 sequences listed in Table C or D, in any combination; or

(iii) sequences having a total of at least one and no more than 5, 4, 3, 2 or 1 amino acid modifications (preferably amino acid substitutions, preferably conservative substitutions) in three CDRs compared to the sequences of point (i) or point (ii) ).

In another embodiment, an antibody or antigen-binding fragment thereof disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the antibody comprises:

(i) six CDR sequences contained in the VH and VL sequences of any of the antibodies listed in Table B;

(ii) the six CDR sequences listed in Table C or D, in any combination; or

(iii) sequences having a total of at least one and no more than 10, 5, 4, 3, 2 or 1 amino acid modifications (preferably amino acid substitutions, preferably conservative substitutions) in six CDRs compared to the sequences of item (i) or item (ii).

In one embodiment, an antibody or antigen-binding fragment thereof disclosed herein includes:

(i) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 84, 85, 86 or 87 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 104;

(ii) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 88, 89 or 90 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 105 or 106;

(iii) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 91, 92 or 93 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 107;

(iv) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 94, 95, 96 or 97 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 108;

(v) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO: 98, 99 or 100 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 set forth in SEQ ID NO: 109; or

(vi) the heavy chain variable region sequences HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO: 101, 102 or 103 and the light chain variable region sequences LCDR1, LCDR2 and LCDR3 set forth in SEQ ID NO: 110.

In a preferred embodiment, the antibody or antigen-binding fragment thereof disclosed herein comprises three complementarity determining regions from the heavy chain variable region (HCDR) and three complementarity determining regions from the variable light chain region (LCDR), where

(i) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 1-5 or 178-181, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 6-10 , HCDR3 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 11-15 or 182-185, LCDR1 contains or consists of an amino acid sequence shown in SEQ ID NO: 16, LCDR2 contains or consists of an amino acid sequence presented in SEQ ID NO: 17, and LCDR3 contains or consists of the amino acid sequence presented in SEQ ID NO: 18;

(ii) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 19-21 or 186-187, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 22-24 , HCDR3 contains or consists of the amino acid sequence shown in SEQ ID NO: 25 and 188, LCDR1 contains or consists of the amino acid sequence shown in SEQ ID NO: 26, LCDR2 contains or consists of the amino acid sequence shown in SEQ ID NO: 27 , and LCDR3 contains or consists of an amino acid sequence selected from the sequences presented in SEQ ID NO: 28-30;

(iii) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 31-33 or 189-190, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 34-37 , HCDR3 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 38-40 or 191-192, LCDR1 contains or consists of an amino acid sequence shown in SEQ ID NO: 41, LCDR2 contains or consists of an amino acid sequence presented in SEQ ID NO: 42, and LCDR3 contains or consists of the amino acid sequence presented in SEQ ID NO: 43;

(iv) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 44-47 or 193-195, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NOs: 48-52 , HCDR3 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 53-56 or 196-198, LCDR1 contains or consists of the amino acid sequence shown in SEQ ID NO: 57, LCDR2 contains or consists of the amino acid sequence presented in SEQ ID NO: 58, and LCDR3 contains or consists of the amino acid sequence presented in SEQ ID NO: 59;

(v) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 60-62 or 199-200, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 6 or 63 -65, HCDR3 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 66-68 or 201-202, LCDR1 contains or consists of the amino acid sequence shown in SEQ ID NO: 69, LCDR2 contains or consists of the amino acid sequence shown in SEQ ID NO: 17, and LCDR3 contains or consists of the amino acid sequence shown in SEQ ID NO: 70; or

(vi) HCDR1 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 71-74 or 203-205, HCDR2 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 22 or 75 -77, HCDR3 contains or consists of an amino acid sequence selected from the sequences shown in SEQ ID NO: 78-80 or 206-207, LCDR1 contains or consists of the amino acid sequence shown in SEQ ID NO: 81, LCDR2 contains or consists of the amino acid sequence shown in SEQ ID NO: 82, and LCDR3 contains or consists of the amino acid sequence shown in SEQ ID NO: 83.

In one preferred embodiment, the antibody or antigen-binding fragment thereof disclosed herein comprises 6 CDR sequences in one of the combinations listed in Table C.

In another preferred embodiment, the antibody or antigen-binding fragment disclosed herein comprises 6 CDR sequences in one of the combinations listed in Table D.

Variable regions of antibodies

A "variable region" or "variable region" is the heavy or light chain domain of an antibody that is involved in the binding of an antibody to its antigen. In the heavy chain variable region (VH) and in the light chain variable region (VL), hypervariable regions (HVRs, also known as complementarity determining regions (CDRs)) with more conserved regions (framework regions (FR)) located between them. Each VH or VL domain consists of three CDRs and four FRs, which are arranged in the following order from the N-terminus to the C-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some cases, antigen-binding specificity may be provided by a single VH or VL domain. Additionally, antibodies that bind to specific antigens can be isolated by screening libraries of complementarity determining VL or VH domains due to the ability of antibody VL or VH domains to bind to antigens (see, e.g., Portolano, S. et al . , J. Immunol 150 (1993) 880-887; Clackson, T. et al. , Nature 352 (1991) 624-628).

Those skilled in the art will recognize that one or more residues in one or both of the variable regions (VH and/or VL) may be modified, e.g., residues in one or more CDRs and/or one or more framework regions, in particular by replacing conserved residues, and the antibody thus modified retains substantially at least one biological property (eg, the ability to bind to an antigen) of the antibody molecule prior to modification. For example, residues in the CDR regions can be mutated to improve one or more characteristics of an antibody related to its antigen binding (eg, affinity). The antigen-binding characteristics or other functional characteristics of the mutated antibody can be studied by in vitro or in vivo assay methods. Preferably, conservative substitutions are made. Preferably, no more than 1, 2, 3, 4, or 5 residue modifications are introduced in the CDR region. In addition, residues in the framework regions can be mutated, for example, to improve antibody performance. For example, one or more residues in the framework regions can be replaced with the corresponding residues of the "germ" sequence through "back mutations".

Another method of modifying the variable region of an antibody known to those skilled in the art is "CDR grafting". Because CDR sequences are responsible for most antigen-antibody interactions, a recombinant antibody variant can be designed to mimic the properties of known antibodies. In such an embodiment, CDR sequence antibodies from known antibodies are "grafted" into the framework regions of other antibodies with differing properties. Accordingly, in one embodiment, the present invention provides an anti-TIGIT antibody or antigen-binding fragment thereof, wherein the antibody or fragment thereof comprises CDR sequences from the heavy and light chain variable regions of one of the antibodies listed in Table B and other framework sequences. . The framework region sequence for replacement can be obtained from public DNA databases containing germline antibody gene sequences, or from published anti-TIGIT antibody sequences. For example, germline DNA encoding heavy and light chain variable region genes can be obtained from the GenBank database. Antibody protein sequences can be matched to protein sequences in a database using sequence similarity analysis tools such as Gapped BLAST. In a preferred embodiment, the framework sequence to be replaced has structural similarity to the framework sequence of the antibody of the present invention selected to be modified, e.g., a framework sequence having a sequence identity of at least 80%, 85%, 90%, 95%, 96%, 97 %, 98%, 99% or more.

In yet another embodiment, the VH and VL sequences from an exemplary antibody of the present invention (one of the antibodies shown in Table B) and other non-TIGIT antibodies (preferably the other antibody shown in Table B) can be "combined and combined into couples" to obtain other antibodies of the present invention that bind to TIGIT. In the case of combination and pairing of such strands, it is preferred that the VH sequence from a particular VH/VL pair is replaced with a structurally similar VH sequence. Similarly, a VL sequence from a particular VH/VL pair is preferably replaced with a structurally similar VL sequence. The binding of such "combined and paired" antibodies to TIGIT can be studied by binding assay methods known to those skilled in the art (eg, ELISA, as well as other assay methods described in the examples).

Thus, in one embodiment, an antibody of the present invention comprises or consists of a heavy chain variable region (VH) sequence of any of the antibodies listed in Table B. In another embodiment, an antibody of the present invention comprises a variant VH sequence.

In another embodiment, an antibody of the present invention comprises or consists of a light chain variable region (VL) sequence of any of the antibodies listed in Table B. In another embodiment, an antibody of the present invention comprises a variant VL sequence.

In yet another embodiment, an antibody of the present invention comprises:

(i) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 84, 85, 86 or 87, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 104, or a variant thereof;

(ii) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 88, 89 or 90, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 105 or 106, or a variant thereof ;

(iii) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 91, 92 or 93, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 107, or a variant thereof;

(iv) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 94, 95, 96 or 97, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 108, or a variant thereof;

(v) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 98, 99 or 100, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 109, or a variant thereof; or

(vi) a VH sequence containing the amino acid sequence shown in SEQ ID NO: 101, 102, or 103, or a variant thereof, and/or a VL sequence containing the amino acid sequence shown in SEQ ID NO: 110, or a variant thereof.

In one embodiment of the invention, in terms of amino acid sequence, the VH sequence variant is at least 80%, 85%, 90%, 92 identical to the reference VH sequence (preferably when compared to the full length sequence or to the CDR1, CDR2 and CDR3 regions). %, 95%, 97%, 98%, 99% or more. In one embodiment of the invention, with respect to the amino acid sequence, the VH sequence variant comprises at least one and no more than 30, 10, 5, 4, 3, 2, 1, or 0 amino acid modifications (preferably amino acid substitutions, and more preferably conservative substitutions). ) compared to the VH reference sequence (preferably when compared to the full length sequence or to the CDR1, CDR2 and CDR3 regions). Preferably, sequence differences do not occur in CDRs.

In a preferred embodiment of the invention, in terms of amino acid sequence, the VL sequence variant is at least 80%, 85%, 90%, 92 identical to the VL reference sequence (preferably when compared to the full length sequence or to the CDR1, CDR2 and CDR3 regions). %, 95%, 97%, 98%, 99% or more. In a preferred embodiment of the invention with respect to the amino acid sequence, the VL sequence variant comprises at least one and no more than 30, 10, 5, 4, 3, 2, 1, or 0 amino acid modifications (preferably amino acid substitutions, and more preferably conservative substitutions). ) compared to the VH reference sequence (preferably when compared to the full length sequence or to the CDR1, CDR2 and CDR3 regions). Preferably, sequence differences do not occur in CDRs.

In a preferred embodiment, an antibody of the present invention comprises or consists of a VH/VL sequence pair from the heavy and light chain variable regions of any of the antibodies listed in Table B. The subject of the present invention is also a variant of an antibody, e.g., a variant having the identity not less than 95-99% or containing no more than 10 amino acid modifications when compared to VH, VL or VH and VL.

In any of the above embodiments, preferably with respect to one or more CDRs (preferably all three CDRs), the heavy chain variable region of the antibody variant comprises no more than 10, preferably no more than 5 (e.g., 3, 2, 1, or 0 ) amino acid modifications (preferably amino acid substitutions, and more preferably conservative substitutions) compared to the reference antibody.

In any of the above embodiments, preferably with respect to one or more CDRs (preferably all three CDRs), the light chain variable region (VL) of an antibody variant comprises no more than 10, preferably no more than 5 (e.g., 3, 2, 1 or 0) amino acid modifications (preferably amino acid substitutions, and more preferably conservative substitutions) compared to the reference antibody.

Heavy and light chains of antibodies

In some embodiments, an antibody of the present invention comprises a heavy chain Fc region, such as an Fc region of immunoglobulins with IgG1, IgG2, or IgG4 isotypes. In one embodiment, an antibody of the present invention comprises an IgG4 Fc region that has a serine-to-proline (S228P) mutation at amino acid position 228 (EU numbering). In another preferred embodiment of the invention, the antibody of the present invention includes the Fc region of IgG4-PAA. The Fc region of IgG4-PAA has a serine-to-proline mutation (S228P) at position 228, a phenylalanine-to-alanine mutation at position 234 (EU numbering), and a leucine-to-alanine mutation at position 235 (EU numbering). Mutation S228P is a mutation in the hinge region of the "tumor" constant region, which is able to reduce or eliminate the heterogeneity of the disulfide bridge between heavy chains. F234A and L235A mutations can further impair the effector function of human IgG4 isotype immunoglobulin (whose effector function is already reduced). In some embodiments, an antibody of the present invention comprises a heavy chain IgG4-PAA Fc region with lysine removed from the C-terminus (des-Lys). In some embodiments, an antibody of the present invention comprises a kappa (κ) light chain constant region, such as a human κ light chain constant region.

In yet another preferred embodiment of the invention, the Fc region comprises an amino acid sequence shown in SEQ ID NO: 177 or an amino acid sequence comprising at least one, two or three, but not more than 20, 10 or 5 amino acid modifications compared to the amino acid sequence, presented in SEQ ID NO: 177, or the sequence is identical to the amino acid sequence presented in SEQ ID NO: 177 at least 95-99%.

In a preferred embodiment, an antibody of the present invention comprises a light chain constant region. In a preferred embodiment, the light chain constant region is a human κ light chain constant region. In yet another preferred embodiment of the invention, the light chain constant region comprises an amino acid sequence as set forth in SEQ ID NO: 209 or an amino acid sequence having at least one, two, or three, but no more than 20, 10, or 5 amino acid modifications compared to the amino acid sequence presented in SEQ ID NO: 209, or the amino acid sequence is identical to the amino acid sequence presented in SEQ ID NO: 209 at least 95-99%.

In some preferred embodiments of the invention, the antibody of the present invention comprises a heavy chain containing an amino acid sequence selected from SEQ ID NOs: 111-130, or an amino acid sequence containing at least one, two or three, but not more than 20, 10 or 5 amino acids modifications compared to the specified sequences, or the amino acid sequence is identical to the specified sequences at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more. Preferably the amino acid modifications do not occur in the CDRs, and more preferably the amino acid modifications do not occur in the variable regions.

In some preferred embodiments, an antibody of the present invention comprises a light chain comprising an amino acid sequence selected from SEQ ID NOs: 137-143, or an amino acid sequence containing at least one, two, or three, but not more than 20, 10, or 5 amino acids. modifications compared to the specified sequences, or the amino acid sequence is identical to the specified sequences at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more. Preferably the amino acid modifications do not occur in the CDRs, and more preferably the amino acid modifications do not occur in the variable regions.

In a preferred embodiment, an antibody of the present invention comprises a heavy chain sequence and/or a light chain sequence selected from:

(a) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 111-114, or a variant thereof, and/or a light chain sequence comprising the amino acid sequence shown in SEQ ID NO: 137, or a variant thereof;

(b) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 115-117, or a variant thereof, and/or a light chain sequence comprising an amino acid sequence set forth in SEQ ID NO: 138 or 139, or a variant thereof;

(c) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 118-120 or a variant thereof, and/or a light chain sequence comprising the amino acid sequence shown in SEQ ID NO: 140 or a variant thereof;

(d) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 121-124, or a variant thereof, and/or a light chain sequence comprising the amino acid sequence shown in SEQ ID NO: 141, or a variant thereof;

(e) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 125-127, or a variant thereof, and/or a light chain sequence comprising the amino acid sequence shown in SEQ ID NO: 142, or a variant thereof; and

(f) a heavy chain sequence comprising an amino acid sequence selected from SEQ ID NO: 128-130 or a variant thereof, and/or a light chain sequence comprising an amino acid sequence shown in SEQ ID NO: 143 or a variant thereof,

where the variant includes an amino acid sequence containing at least one, two or three, but not more than 20, 10 or 5 amino acid modifications or having an identity of at least 80%, 85%, 90%, 92%, 95%, 97%, 98% , 99% or more percent when compared with the corresponding reference sequence. Preferably the amino acid modifications do not occur in the CDRs, and more preferably the amino acid modifications do not occur in the variable regions.

In one embodiment of the invention, residue modifications are made in the constant region of the antibody, for example, to change properties of the antibody, such as effector function.

In some embodiments, the heavy and/or light chain of an anti-TIGIT antibody or fragment disclosed herein further comprises a signal peptide sequence, such as METDTLLLWVLLLWVPGSTG.

Exemplary Antibody Sequences

The subject of the present invention is a fully humanized antibody that specifically binds to TIGIT (eg, human TIGIT), isolated and characterized as described in the examples. The VH and VL sequences from the variable regions of the exemplary antibodies disclosed in this document are listed in Table 3 below. Exemplary antibody CDR sequences are listed in Tables 1 and 2 below. The sequence listing shows the amino acid sequences of the heavy and light chains of the exemplary antibodies disclosed herein and the coding nucleotide sequences of the variable regions (VH and VL) of the exemplary antibodies disclosed herein.

Antibody variants

One aspect of the present invention is any of the antibodies disclosed herein, in particular variants of the exemplary antibodies listed in Table B. In one embodiment, the antibody variant retains at least 60%, 70%, 80%, 90%, or 100% the biological activity (eg, antigen-binding activity) of the antibody prior to modification. In some embodiments of the invention, the modification does not result in the loss of the ability of the antibody variant to bind to the antigen, but may optionally affect its properties, for example, increase binding affinity or other effector functions.

It should be understood that the variable regions of the heavy or light chains or all of the CDRs of an antibody can be modified independently or in combination. In some embodiments, no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications are made to one or more or all three heavy chain CDRs. Preferably the amino acid modification refers to an amino acid substitution, preferably a conservative substitution. In some embodiments, no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications are made to one or more, or all three light chain CDRs. In some embodiments, no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications are made to one or more or all six CDR regions. Preferably the amino acid modification refers to an amino acid substitution, preferably a conservative substitution. In some embodiments, the antibody variant has an amino acid sequence that is at least 80%, 85%, 90%, 95%, 99%, or more percent identical to the target region of the reference antibody sequence. For example, in one embodiment, an antibody of the present invention is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more percent identical to a reference antibody. (eg, one of the antibodies listed in Table 3) for three heavy chain CDRs. In one embodiment, an antibody of the present invention is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more percent identical to a reference antibody (e.g. , one of the antibodies listed in Table 3) for three light chain CDRs. In another embodiment, an antibody of the present invention is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more percent identical to a reference antibody (e.g. , one of the antibodies listed in Table 3) for six CDRs. In another embodiment, an antibody of the present invention is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than percent identical to the reference antibody (for example, one of the antibodies listed in Table 3) in relation to the variable region of the heavy chain. In another embodiment, an antibody of the present invention is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than percent identical to the reference antibody (for example, one of the antibodies listed in Table 3) in relation to the variable region of the light chain. In another embodiment, an antibody of the present invention is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than percent identical to the reference antibody (for example, one of the antibodies listed in Table 3) in relation to the variable regions of the heavy and (or) light chains.

In addition, modifications can be made to the Fc region of an antibody. Fc region modifications may be made independently or in combination with the framework and/or CDR modifications described above. The Fc region can be modified, for example, to alter one or more functions of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen-dependent cytotoxicity. In addition, the antibody of the present invention may be chemically modified (eg, linked to PEG) or its glycosylation profile may be altered.

In certain embodiments, the Fc region may include an Fc region having one or more amino acid substitutions that increase antibody-dependent cellular cytotoxicity (ADCC), such as substitutions at positions 298, 333, and/or 334 (EU residue numbering) of the Fc region. In some embodiments, the Fc region can also be modified to alter (i.e. increase or decrease) C1q binding and/or complement dependent cytotoxicity (CDC) ( see e.g. US 6,194,551, WO 99/51642 and Idusogie, EE et al. , J Immunol 164 (2000) 4178-4184).

In other embodiments, the Fc region may be modified to increase or decrease the extent of its glycosylation and/or change its glycosylation profile. A convenient way to add or remove glycosylation sites in the Fc region is to add or remove one or more glycosylation sites by changing the amino acid sequence. For example, one or more amino acids can be substituted at those sites to remove one or more glycosylation sites and thereby eliminate glycosylation at those sites. Antibodies with an altered glycosylation profile can be generated, such as low-fucosylated or non-fucosylated antibodies with reduced fucosyl residues, or antibodies with increased structures with bisected GlcNac. It has been shown that such altered glycosylation profiles can increase the ADCC of antibodies. This document also contemplates antibody variants having at least one galactose residue in an Fc-linked oligosaccharide. In antibody variants, CSC function can be enhanced.

In certain embodiments, the present invention also provides antibody variants having some but not all effector functions, making them desirable candidates for some applications in which the in vivo half-life of antibodies is important, while other effector functions (e.g., complement binding ability or ADCC) are unnecessary or harmful. For example, the Fc region may contain a mutation that eliminates or reduces effector functions, in particular, the Fc region of human IgG1 may have mutations P329G and (or) L234A and L235A, or the Fc region of human IgG4 may have mutations P329G and (or) S228P and L235E.

In certain embodiments of the invention, the present invention may relate to the production of antibodies modified by cysteine engineering, such as "sulfo-MAb", in which one or more antibody residues are replaced by cysteine residues. For example, the number of cysteine residues in the hinge region of an antibody can be altered, for example to facilitate assembly of light and heavy chains, or to increase or decrease the stability of the antibody. See, for example, US Pat. No. 5,677,425.

In certain embodiments, the antibodies provided herein may be further modified to include other non-protein elements. Suitable elements for modifying antibodies include, but are not limited to, water-soluble polymers. An example of a water-soluble polymer is, inter alia, polyethylene glycol (PEG), which is administered, for example, to increase the half-life of an antibody (eg, from blood serum). Methods for PEGylating proteins are known to those skilled in the art and can be applied to the antibodies of the present invention. See, for example, EP 0154316 and EP 0401384.

II. Polynucleotides, vectors and host organisms

The subject of the present invention is a nucleic acid encoding any of the above antibodies to TIGIT or fragments thereof, as well as a vector containing such a nucleic acid. In one embodiment of the invention, the vector is an expression vector. In addition, the subject of the invention is a host cell containing such nucleic acid or vector. In one embodiment of the invention, the host cell is eukaryotic. In another embodiment, the host cell is selected from a yeast cell and a mammalian cell (eg, 293 cells or CHO cells). In yet another embodiment of the invention, the host cell is prokaryotic.

One aspect of the present invention is a nucleic acid encoding any of the above antibodies to TIGIT or fragments thereof. The nucleic acid may include a nucleic acid encoding the amino acid sequences of the light chain variable regions and/or the heavy chain variable regions of antibodies, or a nucleic acid encoding the amino acid sequences of the light chains and/or heavy chains of antibodies. Exemplary nucleic acid sequences encoding heavy chain variable regions include nucleic acid sequences that are at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% , 98% or 99% identical to the nucleic acid sequences selected from SEQ ID NO: 150-169, or include nucleic acid sequences selected from SEQ ID NO: 150-169. Exemplary nucleic acid sequences encoding light chain variable regions include nucleic acid sequences that are at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% , 98% or 99% identical to the nucleic acid sequences selected from SEQ ID NO: 170-176, or include nucleic acid sequences selected from SEQ ID NO: 170-176. The polypeptides encoded by the polynucleotides may exhibit the ability to bind to an antigen (bind to TIGIT) when expressed in a suitable expression vector.

The present invention also provides a polynucleotide encoding at least one CDR, and typically all three CDRs, from the heavy chain (VH) or light chain (VL) sequence of the TIGIT-binding antibodies described above. In some other embodiments, the polynucleotide encodes the entire or substantially the entire variable region sequence of the heavy chains and/or light chains of the TIGIT-binding antibodies described above.

Those skilled in the art will appreciate that the amino acid sequence of each antibody or polypeptide may be encoded by different nucleotide sequences due to the degeneracy of the codon.

In a preferred embodiment, the nucleic acid encoding an antibody of the present invention further comprises a nucleotide sequence encoding a heavy chain Fc region, such as the Fc region sequence shown in SEQ ID NO: 177 or a sequence substantially identical thereto.

In a preferred embodiment of the invention, the nucleic acid encoding the antibody of the present invention further comprises a nucleotide sequence encoding a light chain constant region sequence, such as the sequence shown in SEQ ID NO: 209 or a sequence substantially identical thereto.

Using methods known to those skilled in the art, polynucleotide sequences can be generated by de novo solid phase DNA synthesis or by PCR mutagenesis of existing sequences (e.g., the VH coding DNA sequence shown in SEQ ID NO: 150-169 or VL- the DNA coding sequence shown in SEQ ID NO: 170-176) encoding TIGIT-binding antibodies or antigen-binding fragments thereof.

In one embodiment of the invention, there are one or more vectors containing the nucleic acid of the present invention. In one embodiment of the invention, the vector is an expression vector, such as a eukaryotic expression vector. The vector includes, but is not limited to, a virus, plasmid, cosmid, lambda phage, or yeast artificial chromosome (YAC). In a preferred embodiment of the invention, the expression vector of the present invention is the pTT5 expression vector.

In one embodiment of the invention, there is a host cell containing a vector. A suitable host cell for cloning or expression of an antibody-encoding vector includes a prokaryotic or eukaryotic cell as described herein. For example, antibodies can be produced in bacteria, particularly when glycosylation or Fc-mediated effector functions are not required. Expression of antibody fragments and polypeptides in bacteria is described, for example, in US Pat. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes the expression of antibody fragments in E. coli . Once expressed, the antibodies in the soluble fraction can be isolated from the bacterial cell mass and further purified.

In one embodiment of the invention, the host cell is eukaryotic. In another embodiment of the invention, the host cell is selected from a yeast cell, a mammalian cell, and other cells suitable for producing an antibody or antigen-binding fragment thereof. For example, suitable hosts for cloning or expression of vectors encoding antibodies are eukaryotic microorganisms such as filamentous fungi or yeasts. For example, fungal or yeast strains in which the glycosylation pathway is "humanized" allow the production of antibodies with a partially or fully human glycosylation profile. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004), as well as Li et al. , Nat. Biotech. 24:210-215 (2006). Host cells suitable for the expression of glycosylated antibodies can also be derived from multicellular organisms (invertebrates and vertebrates). Cells of vertebrate organisms can also be used as hosts. For example, a mammalian cell line modified to grow in suspension can be used. Examples of useful mammalian host cell lines are monkey kidney CV1 cell line transformed with SV40 virus (COS-7), human embryonic kidney cell line (HEK 293 or 293 cells as described, for example, in Graham et al ., J. Gen. Virol 36:59 (1977)) and the like. Other useful mammalian host cell lines are Chinese hamster ovary (CHO) cells, including DHFR ^- CHO cells (Urlaub et al ., Proc. Natl. Acad. Sci. USA 77:216 (1980)), and myeloma cell lines such like Y0, NS0 and Sp2/0. Reviews of certain mammalian host cell lines suitable for antibody production are, for example, Yazaki and Wu, Methods in Molecular Biology , Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

III. Obtaining antibodies

In one embodiment of the invention, there is a method for producing an anti-TIGIT antibody, wherein the method comprises culturing host cells containing the nucleic acid encoding the antibody under conditions suitable for expressing the antibody as described above, and optionally isolating the antibody from the host cells (or host cell culture medium). To generate an anti-TIGIT antibody by recombinant DNA, a nucleic acid encoding an antibody (eg, the antibody described above) is isolated and inserted into one or more vectors for further cloning and/or expression in host cells. Such nucleic acid can be easily isolated and sequenced using conventional procedures (eg, using oligonucleotide probes capable of specifically binding to genes encoding antibody heavy and light chains).

IV. Analysis

Anti-TIGIT antibodies of the present invention can be identified, screened, or characterized in terms of their physical/chemical properties and/or biological activity using a variety of assays known to those skilled in the art.

In one aspect of the invention, the antibodies of the present invention are analyzed for antigen-binding activity. For example, the ability to bind to human TIGIT can be determined by methods known to those skilled in the art such as ELISA, Western blotting, and the like, or by the illustrative methods disclosed in the examples provided herein. For example, a flow cytometric assay can be performed where antibodies react with a cell line expressing human TIGIT, such as CHO cells transfected to express human TIGIT on the cell surface. The flow cytometry method can be used for other cells, including T cells expressing native TIGIT. Alternatively, biological optical interferometry using recombinant TIGIT proteins can be used to determine antibody binding, including binding kinetics (eg, K _D ). For example, in some embodiments of the invention, an adapted Fortebio affinity assay method is used.

In another aspect of the invention, a competition assay may be used to identify an antibody that competes for TIGIT binding to any of the antibodies disclosed herein. In certain embodiments, such a competing antibody binds to the same or overlapping epitope (eg, a linear or conformational epitope) as any of the anti-TIGIT antibodies disclosed herein. An exemplary method for determining the position of an epitope to which an antibody binds is described in detail in Morris (1996) "Epitope Mapping Protocols", Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

The subject of the present invention is also a method for the determination of antibodies to TIGIT with biological activity. Biological activity may include, for example, binding to TIGIT (e.g., human TIGIT), blocking TIGIT (e.g., human TIGIT) binding to CD155 molecules, blocking TIGIT-mediated inhibition signaling, increasing IL-2 production in T cells, and/or suppression of tumor growth. For example, the ability of antibodies to suppress tumor growth is tested in an in vivo tumor suppression model (see, for example, Example 6). Antibodies having such in vivo and/or in vitro biological activity are also described in this document.

Those of skill in the art will appreciate that any of the assays mentioned above can be carried out using the immunoconjugates or multispecific antibodies of the present invention in place of and in addition to anti-TIGIT antibodies.

V. Polyspecific antibodies

Another aspect of the present invention is a polyspecific (including bispecific) antibody molecule that specifically binds to TIGIT, preferably human TIGIT. In one embodiment, the antibodies (or antigen-binding fragments thereof) of the present invention, when referred to as multispecific antibodies, specifically bind to TIGIT as the first antigen. In yet another embodiment, the polyspecific antibodies further have the ability to specifically bind to a second antigen, or, in yet another embodiment of the invention, have the ability to specifically bind to a second and third antigen. In yet another embodiment of the invention, the polyspecific antibodies are bispecific antibodies.

In one embodiment, the binding specificity depends on the "binding site" or "antigen-binding site" of the antibody (the region of the antibody molecule that actually binds to the antigen). In a preferred embodiment, the antigen-binding site is formed by a VH/VL pair consisting of a light chain variable region (VL) and a heavy chain variable region (VH) of an antibody. Thus, in one embodiment of the invention, "multispecific" antibodies contain at least two antigen-binding sites, each of which can bind to different epitopes of the same antigen or to different epitopes of different antigens.

Polyspecific antibodies and methods for their preparation are described, for example, in patents WO 2009/080251, WO 2009/080252, WO 2009/080253, WO 2009/080254, WO 2010/112193, WO 2010/115589, WO 2010/136172/WO 2010 145792 and WO 2010/145793.

VI. Immunoconjugates

Another aspect of the present invention is an immunoconjugate obtained by conjugating the antibodies of the present invention with a heterologous molecule. In one embodiment of the invention, an immunoconjugate is prepared by conjugating the antibodies (or antigen-binding fragments thereof) of the present invention to a therapeutic or diagnostic agent. In some embodiments, the antibodies of the present invention may be conjugated to a heterologous molecule in the form of full length antibodies or antibody fragments. For example, antibodies are conjugated as Fab fragments, Fab' fragments, F(ab)' ₂ fragments, single chain scFab antibodies, single chain scFvs, or other fragments.

In other embodiments, the antibodies of the present invention are conjugated to a therapeutic molecule. Linkers can be used to covalently link antibodies to a therapeutic molecule. Suitable linkers include chemical or peptide linkers.

In other embodiments, the antibodies of the present invention may be conjugated to a diagnostic or detectable agent. Such conjugates may be used as part of a clinical trial method (eg, to determine the efficacy of a particular therapy), to monitor or predict the onset, development, progression, and/or severity of a disease or disorder. Such diagnosis or detection can be accomplished by binding the antibodies to the agent to be detected.

VII. Pharmaceutical compositions and pharmaceutical preparations

The subject of the present invention is also a composition (including a pharmaceutical composition or a pharmaceutical preparation) comprising anti-TIGIT antibodies or immunoconjugates thereof, or polyspecific antibodies, as well as a composition comprising a polynucleotide encoding anti-TIGIT antibodies or immunoconjugates thereof, or polyspecific antibodies. Such compositions may optionally further include suitable pharmaceutical excipients, such as a pharmaceutical carrier, pharmaceutical excipient, and the like, known to those skilled in the art, including buffers.

In one embodiment of the invention, the composition further comprises a second therapeutic agent. The second therapeutic agent may be selected from the group consisting of, but not limited to, anti-PD-1 antibodies and anti-PD-L1 antibodies. In a preferred embodiment, the second therapeutic agent is a PD-1 antagonist, in particular an anti-PD-1 antibody.

The pharmaceutical carrier applicable to the present invention may be a sterile liquid such as water or oil, including liquids derived from petroleum, animals, plants, or synthetically derived such as peanut oil, soybean oil, liquid paraffin, sesame oil, etc. P. In case the pharmaceutical composition is administered intravenously, the preferred carrier is water. Saline solutions, aqueous dextrose and glycerol solutions can also be used as liquid carriers, in particular for injection solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, rye malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene, glycol, water, ethanol and etc. The uses and uses of excipients are described in Handbook of Pharmaceutical Excipients , 5th ed., RC Rowe, PJ Seskey and SC Owen, Pharmaceutical Press, London, Chicago. The composition may further include a small amount of wetting agents or emulsifiers, or, if desired, a buffer to maintain the pH. The composition may be in the form of a solution, suspension, emulsion, tablet, pill, capsule, powder, sustained release formulation, and the like. Oral preparations may include standard carriers such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate and saccharin.

The pharmaceutical preparation of the present invention may be prepared by mixing the anti-TIGIT antibodies, immunoconjugates or polyspecific antibodies of the present invention of the desired purity with one or more optional pharmaceutical excipients, preferably in the form of a lyophilized preparation or an aqueous solution ( Remington 's Pharmaceutical Sciences , 16- e ed., Osol, A. eds. (1980)).

An exemplary lyophilized antibody preparation is described in US Pat. No. 6,267,958. The aqueous preparation of the antibodies includes those described in US Pat.

The pharmaceutical composition or formulation of the present invention may further comprise one or more other active ingredients required for the particular indication to be treated, and preferably the active ingredients have complementary properties without adversely affecting each other. For example, it may be desirable to further include other active ingredients with antitumor activity, such as PD-1 binding antagonists or PD-L1 binding antagonists, for example an anti-PD-1 antibody or an anti-PD-L1 antibody. The active ingredients are appropriately combined in amounts that are effective for the intended purpose.

Can be obtained drug with prolonged release. An example of a sustained release formulation is a semi-permeable solid hydrophobic polymer matrix containing an antibody. The matrix is made in the form of a profiled product, such as a film or a microcapsule.

Other components of the pharmaceutical formulation are described in WO 2015/153513.

VIII. Combined drugs

One aspect of the present invention is also a combination preparation comprising antibodies or antigen-binding fragments thereof, bispecific antibodies or immunoconjugates of the present invention and one or more therapeutic agents (for example, a chemotherapeutic agent, other antibodies, a cytotoxic agent, a vaccine, an anti-infective active agent) . The combination preparation of the present invention can be used in the therapeutic method disclosed in this document.

In some embodiments, there is a combination preparation wherein the other therapeutic agents refer to, for example, a therapeutic agent such as an antibody that has the ability to stimulate an immune response and thereby further enhance, stimulate, or increase the immune response in a subject. In some embodiments, the other antibodies refer to, for example, an anti-PD-1 antibody or an anti-PD-L1 antibody.

In some embodiments, the combined preparation is used to prevent or treat a neoplastic disease. In some embodiments, the tumor is a cancer, such as a gastrointestinal cancer (eg, stomach cancer, rectal cancer, colon cancer, and colorectal cancer) or a skin cancer (eg, malignant melanoma). In some embodiments, the combination preparation is used to prevent or treat an infection, such as a bacterial infection, a viral infection, a fungal infection, a protozoan infection, and the like.

IX. Method of therapy and use of antibodies

In this document, the terms "individual" and "subject" can be used interchangeably and refer to a mammal. Mammals include, but are not limited to, domestic animals (eg, cows, sheep, cats, dogs, and horses), primates (eg, primates and non-human primates such as monkeys), rabbits, and rodents (eg, mice and rats). In particular, the subject is a person.

As used herein, the term "treatment" refers to a clinical intervention to alter the natural course of a disease in a subject being treated. Desirable therapeutic effects include, but are not limited to, preventing the onset or recurrence of diseases, alleviating symptoms, improving any direct or indirect disease outcomes, preventing metastasis, slowing disease progression, improving or alleviating the condition, and improving prognosis.

In one aspect, the present invention relates to a method for enhancing the immune response of a subject's body, which method includes administering to the subject an effective amount of any of the anti-TIGIT antibodies or fragments thereof described herein, an immunoconjugate or polyspecific antibody comprising antibodies or fragments thereof, or a pharmaceutical compositions. In some embodiments, the anti-TIGIT antibodies or antigen-binding fragments thereof of the present invention are administered to a subject with a tumor to stimulate an anti-tumor immune response. In other embodiments, the antibodies or antigen-binding fragments thereof of the present invention are administered to a subject with an infection to stimulate an immune response against the infection.

In another aspect, the present invention relates to a method of treating a neoplastic disease, such as malignancy, in a subject, where this method includes administering to the subject an effective amount of any of the antibodies to TIGIT or fragments thereof described in this document, an immunoconjugate or a multispecific antibody, including antibodies or their fragments, or pharmaceutical composition. The malignancy may be early, intermediate or late stage, or metastatic malignancy.

It has been shown that TIGIT is highly expressed on the surface of human tumor-infiltrating CD8 ⁺ T cells. In some embodiments, the method of the present invention is used to treat a cancer, in particular a solid tumor infiltrated by TIGIT-expressing tumor-infiltrating lymphocytes.

In one embodiment of the invention, the oncological disease is a malignant neoplasm of the gastrointestinal tract, such as colon cancer.

In some embodiments, the tumor or tumor cell may be selected from colorectal neoplasm, ovarian neoplasm, pancreatic neoplasm, lung neoplasm, liver neoplasm, breast neoplasm, kidney neoplasm, prostate neoplasm, gastrointestinal neoplasm, melanoma, cervical neoplasm , neoplasms of the bladder, glioblastomas, as well as neoplasms of the head and neck. In some embodiments, the cancer may be selected from colorectal cancer, ovarian cancer, pancreatic cancer, lung cancer, liver cancer, breast cancer, kidney cancer, prostate cancer, gastrointestinal cancer, melanoma, cervical cancer, bladder cancer, glioblastoma, and malignant neoplasms of the head and neck.

In some embodiments of the invention, there is a method of treating a neoplastic disease (eg, malignancy), where this method includes the introduction of the subject of antibodies to TIGIT of the present invention and antagonistic antibodies to PD-1. In some embodiments, the method is used to treat a neoplastic disease in which the tumor coexpresses TIGIT and PD-1, such as TIGIT and PD-1 coexpressing melanoma (Chauvin et al . (2015) J. Clin . Invest. 125:2046), non-small cell lung cancer (NSCLC) and renal cell carcinoma (RCC).

In another aspect, the present invention provides a method of treating an infectious disease, such as a chronic infection, in a subject, wherein the method comprises administering to the subject an effective amount of any of the anti-TIGIT antibodies or fragments thereof described herein, an immunoconjugate, or a multispecific antibody comprising the antibodies or antibody thereof. fragments, or pharmaceutical composition. In one embodiment of the invention, the infection is a viral infection.

In one embodiment of the invention, the infectious disease results from a viral infection. Some examples of pathogenic viruses are hepatitis viruses (A, B and C), influenza viruses (A, B and C), HIV, herpes viruses (for example, VZV, HSV-1, HAV-6, HSV-II, CMV and Epstein virus -Barr), adenoviruses, flaviviruses, echoviruses, rhinoviruses, Coxsackieviruses, coronaviruses, respiratory syncytial viruses, mumps viruses, rotaviruses, measles viruses, rubella viruses, parvoviruses, vaccinia viruses, human T-cell leukemia viruses (HTLV), viruses dengue, papillary carcinomas, molluscum viruses, polioviruses, rabies viruses, JC viruses and arthropod-borne encephalitis viruses.

In some embodiments of the invention, the method described herein further includes applying to the subject one or more therapeutic interventions in combination with each other (for example, a therapeutic method and (or) other therapeutic agents). In some embodiments, the therapeutic method includes surgery and/or radiation therapy.

In some embodiments, the T-cell response can be stimulated by the combination of the anti-TIGIT antibodies of the present invention and one or more therapeutic agents. In some embodiments of the invention, in addition to administering the antibodies of the present invention, the method of the present invention further comprises administering at least one additional antibody with immunostimulatory activity, e.g. humanized, chimeric or humanized.

In other embodiments, other therapeutic agents that can be used in combination with the antibodies of the present invention are PD-1 binding antagonists or PD-L1 binding antagonists. Alternate names for "PD-1" include CD279 and SLEB2. Alternate names for "PD-L1" include B7-H1, B7-4, CD274, and B7-H. In some embodiments, PD-1 and PD-L1 are human PD-1 and human PD-L1, respectively. In some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its ligand/binding partner. In one specific aspect, the PD-1 ligand/binding partner is PD-L1. In another embodiment, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In one specific aspect, the PD-L1 binding partner is PD-1. The antagonist may be an antibody, antigen binding fragment, immunoadhesin, fusion protein, or oligopeptide. In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody (eg, a human antibody, a humanized antibody, or a chimeric antibody). In some embodiments, the anti-PD-1 antibody is selected from the following list: IBI308 (sintilumab monoclonal antibody, WO2017/025016A1), MDX-1106 (nivolumab, OPDIVO), Merck 3475 (MK-3475, pembrolizumab, KEYTRUDA), and CT-011 (pidilizumab). In some embodiments, the anti-PD-1 antibody is MDX-1106. In some embodiments, the anti-PD-1 antibody is nivolumab (CAS Registry Number: 946414-94-4). In a preferred embodiment, the anti-PD-1 antibody is "Antibody C" as described herein.

In some other embodiments, anti-TIGIT antibodies or fragments thereof, alone or in combination with a PD-1/PD-L1 binding antagonist, may be used in combination with one or more therapeutic modalities, such as therapeutic modalities and/or other therapeutic agents. In some embodiments of the invention, therapeutic methods include surgery (for example, tumor resection), radiation therapy (for example, external beam radiation therapy, including three-dimensional conformal radiation therapy, in which the radiation zone is specially defined), partial radiation (for example, radiation aimed at preselected target or organ), focused irradiation, and the like.

In some embodiments, the anti-TIGIT antibodies or antigen-binding fragments thereof of the present invention may be used in combination with chemotherapy or a chemotherapeutic agent. In some embodiments, the anti-TIGIT antibodies or antigen-binding fragments thereof of the present invention may be used in combination with radiation therapy or a radiotherapeutic agent. In some embodiments, the anti-TIGIT antibodies or antigen-binding fragments thereof of the present invention may be used in combination with a targeted therapy or a targeted therapeutic agent. In some embodiments, the anti-TIGIT antibodies or antigen-binding fragments thereof of the present invention may be used in combination with immunotherapy or an immunotherapeutic agent, such as a monoclonal antibody.

An antibody of the present invention (pharmaceutical composition or immunoconjugate comprising the same or any other therapeutic agents) may be administered by any suitable route, eg, parenteral, intrapulmonary, intranasal, and intralesional if topical treatment is required. Parenteral administration includes intramuscular, intravenous, intra-arterial, intraperitoneal or subcutaneous administration. Drugs may be administered by any suitable route, in particular by injection, such as intravenous or subcutaneous injection, depending to some extent on whether the treatment is short-term or long-term. The invention covers various administration regimens, including, but not limited to, single administration or multiple administrations, bolus injections, and fractional infusions at different time points.

In the case of use for the prevention and treatment of diseases, the appropriate dose of antibodies of the present invention (when used alone or in combination with one or more therapeutic agents) will depend on the type of disease being treated, the type of antibodies, the severity and progression of the disease, the purpose of application ( prophylactic or therapeutic), previous therapy, the patient's clinical history, antibody response, and the opinion of the attending physician. Acceptable use of the antibody in a patient is a single injection or a series of multiple injections.

In the methods described above, the composition, multispecific antibody or immunoconjugate of the present invention can be used instead of the antibody or antigen-binding fragment described in this document. Alternatively, in these methods, the composition, polyspecific antibody or immunoconjugate of the present invention may also be used after the application of the antibody or antigen-binding fragment described herein.

Another aspect of the present invention is the use of an anti-TIGIT antibody, a composition, an immunoconjugate, and a multispecific antibody of the present invention for the preparation of medicaments useful in the methods described above (eg, for treatment).

X. Methods and compositions for diagnosis and detection (detection)

Another aspect of the present invention relates to a method and kit for detecting TIGIT in a sample, where this method includes the following steps: (a) bringing the sample into contact with the antibody or antigen-binding fragment or immunoconjugate disclosed in this document; and (b) detecting a complex formed between the antibody or antigen-binding fragment or immunoconjugate and the TIGIT protein. In some embodiments, the sample is from a cancer patient, such as a skin cancer patient. The detection may be carried out in vitro or in vivo .

The term "detection (detection)" as used herein includes quantitative or qualitative detection, and exemplary detection options may include detection by immunohistochemistry, immunocytochemistry, flow cytometry (e.g., FACS), magnetic beads associated with antibody molecules, solid-phase immunoassay (ELISA); and PCR methods (eg, PCR-RT). In certain embodiments of the invention, the biological sample is blood, serum, or other liquid sample of biological origin. In certain embodiments of the invention, the biological sample includes cells or tissues. In some embodiments, the biological sample originates from a proliferative or malignant lesion. In certain embodiments of the invention, the TIGIT to be detected is a human TIGIT.

In one embodiment, an anti-TIGIT antibody is used to select a subject suitable for treatment with an anti-TIGIT antibody, eg TIGIT is a biomarker for selecting a subject. In one embodiment, an antibody of the present invention can be used for the diagnosis of malignancies and tumors, for example, to evaluate (e.g., monitor) treatment or progression, diagnose and/or staging a disease (e.g., a hyperproliferative or malignant disease) described in this document, at the subject.

In certain embodiments of the invention, there is a labeled anti-TIGIT antibody. The label includes, but is not limited to, a label or group (e.g., a fluorescent label, a chromophore label, an electron-dense label, a chemiluminescent label, and a radioactive label) that can be detected directly, as well as a group that can be detected indirectly, such as an enzyme or ligand that can be detected, for example, by enzymatic reaction or molecular interaction. Examples of labels include but are not limited to ³² P, ¹⁴ C, ¹²⁵ I, ³ H and ¹³¹ I radioisotopes, fluorophores (e.g. rare earth chelates or fluorescein) and their derivatives, rhodamine and its derivatives, dansyl, umbelliferone, luciferase (e.g. luciferase firefly and bacterial luciferase (U.S. Patent No. 4,737,456)), fluorescein, 2,3-dihydrophthalazinedione, horseradish peroxidase (HRP), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, carbohydrate oxidase (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase ), heterocyclic oxidase (for example, uricase and xanthine oxidase), enzymes that oxidize dye precursors in the presence of hydrogen peroxide (for example, HRP, lactoperoxidase, or microperoxidase), biotin/avidin, spin labels, phage labels, stable free radicals, etc.

In order to facilitate understanding of the present invention, the following examples are given below. These examples are not intended to diminish the scope of the present invention and should not be interpreted as such.

Table 1. CDR sequences of exemplary antibodies of the present invention

Antibody HCDR1 SEQ ID
NO: HCDR2 SEQ ID
NO: HDDR3 SEQ ID
NO: LCDR1 SEQ ID
NO: LCDR2 SEQ ID
NO: LCDR3 SEQ ID
NO: Numbering of positions according to Kabat H27-35B H50-65 H93-102 L24-34 L50-56 L89-97 ADI-27238 YTF T SYYM S one II N PSGG S TS Y A Q KFQG 6 ARAR Y PSSWPYG M DV eleven RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30263 YTF L SYYM N 2 II D PSGG R TS Y A Q KFQG 7 ARAR Y PSSWPYG T DV 12 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30267 YTF R SYYM S 3 II D PSGG R TS F A Q KFQG eight ARAR Y PESWPYG M DV 13 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30268 YTF G SYYM G 4 II D PSGG R TS Y A R KFQG nine ARAR T PSSWPYG M DV fourteen RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen Consensus Sequence YTFX ₁ SYYMX ₂
(Where X ₁ is selected from T, L, R, G and residues that provide their conservative replacement, and X ₂ is selected from S, N, G and residues that provide their conservative replacement) five IIX ₁ PSGGX ₂ TSX ₃ AX ₄ KFQG
(where X ₁ is selected from N, D and residues that provide their conservative replacement, X ₂ is selected from S, R and residues that provide their conservative replacement, X ₃ is selected from Y, F and residues that provide their conservative replacement, and X ₄ selected from Q, R and residues providing their conservative substitution) 10 ARARX ₁ PSSWPYGX ₂ DV
(Where X ₁ is selected from Y, T and residues that provide their conservative replacement, and X ₂ is selected from M, T and residues that provide their conservative replacement) fifteen RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-27243 GSI S SS R YYWG nineteen SIY Y SG S TYYNPSLKS 22 ARDGLYHTPEYFQH 25 RASQSVSSYLA 26 DASNRAT 27 QQRFA F PYT 28 ADI-30302 GSI G SS Q YYWG twenty SIY R SG G TYYNPSLKS 23 ARDGLYHTPEYFQH 25 RASQSVSSYLA 26 DASNRAT 27 QQRFA F PYT 28 ADI-30336 GSI S SS R YYWG nineteen SIY Y SG S TYYNPSLKS 22 ARDGLYHTPEYFQH 25 RASQSVSSYLA 26 DASNRAT 27 QQRFA H PYT 29 Consensus Sequence GSIX ₁ SSX ₂ YYWG
(where X ₁ is selected from S, G and residues that provide their conservative replacement, and X ₂ is selected from R, Q and residues that provide their conservative replacement) 21 SIYX ₁ SGX ₂ TYYNPSLKS
(where X ₁ is selected from Y, R and residues that provide their conservative replacement, and X ₂ is selected from S, G and residues that provide their conservative replacement) 24 ARDGLYHTPEYFQH 25 RASQSVSSYLA 26 DASNRAT 27 QQRFAX ₁ PYT
(Where X ₁ is selected from F, H and residues providing their conservative substitution) thirty ADI-27278 FT FSS YSMN 31 YI SG SS S TI Y 34 ARHRI AD SPSRAFDI 38 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-30293 FT FSS YSMN 31 YI -S SS G TI N YADSVKG 35 ARHRI AD SPSRAFDI 38 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-30296 FT IGG YSMN 32 YI -S SS S TI H YADSVKG 36 ARHRI GR SPSRAFDI 39 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 Consensus Sequence FTX ₁ X ₂ X ₃ YSMN
(Where X ₁ is selected from F, I and residues that provide their conservative replacement, X ₂ is selected from S, G and residues that provide their conservative replacement, and X ₃ is selected from S, G and residues that provide their conservative replacement) 33 YIX ₁ X ₂ SSX ₃ TIX ₄ YADSVKG
(Where X ₁ is selected from deletion, S and residues that provide its conservative replacement, X ₂ is selected from G, S and residues that provide their conservative replacement, X ₃ is selected from S, G and residues that provide their conservative replacement, and X ₄ selected from Y, N, H and residues providing their conservative substitution) 37 _ARHRIX 1X2 _SPSRAFDI
(where X ₁ is selected from A, G and residues that provide their conservative replacement, and X ₂ is selected from D, R and residues that provide their conservative replacement) 40 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-27291 FTF SS Y A MS 44 A ISGSG GS T YY ADSVKG 48 AKDPGT DSS GYYYIWRY 53 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30283 FTF SP Y G MS 45 S ISGSG GR T YY ADSVKG 49 AKDPGT DSS GYYYIWRY 53 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30286 FTF GP Y G MS 46 A ISGSG AS T WY ADSVKG fifty AKDPGT HYS GYYYIWRY 54 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30288 FTF GP Y G MS 46 A ISGSG AS T WH ADSVKG 51 AKDPGT DST GYYYIWRY 55 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 Consensus Sequence FTFX ₁ X ₂ YX ₃ MS
(Where X ₁ is selected from S, G and residues that provide their conservative replacement, X ₂ is selected from S, P and residues that provide their conservative replacement, and X ₃ is selected from A, G and residues that provide their conservative replacement) 47 X ₁ ISGSGX ₂ X ₃ TX ₄ X ₅ ADSVKG
(Where X ₁ is selected from A, S and residues that provide their conservative replacement, X ₂ is selected from G, A and residues that provide their conservative replacement, X ₃ is selected from S, R and residues that provide their conservative replacement, X ₄ is selected from Y, W and residues providing their conservative replacement, and X ₅ is selected from Y, H and residues providing their conservative replacement) 52 AKDPGTX ₁ X ₂ X ₃ GYYYIWRY
(Where X ₁ is selected from D, H and residues that provide their conservative replacement, X ₂ is selected from S, Y and residues that provide their conservative replacement, and X ₃ is selected from S, T and residues that provide their conservative replacement) 56 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-27297 YTFT S YYMH 60 II N PS G GST S YAQKFQG 6 ARDHDI AA AGRLADY 66 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-30272 YTFT E YYMH 61 II S PS A GST S YAQKFQG 63 ARDHDI AA AGRLADY 66 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-30278 YTFT E YYMH 61 II S PS A GST K YAQKFQG 64 ARDHDI RL AGRLADY 67 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 Consensus Sequence YTFTX ₁ YYMH
(Where X ₁ is selected from S, E and residues providing their conservative substitution) 62 IIX ₁ PSX ₂ GSTX ₃ YAQKFQG
(Where X ₁ is selected from N, S and residues that provide their conservative replacement, X ₂ is selected from G, A and residues that provide their conservative replacement, and X ₃ is selected from S, K and residues that provide their conservative replacement) 65 _ARDHDIX 1X2 _AGRLADY
(Where X ₁ is selected from A, R and residues that provide their conservative replacement, and X ₂ is selected from A, L and residues that provide their conservative replacement) 68 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-27301 GSI S SS S YYWG 71 SIYYSGST Y YNPS L KS 22 AREAGR GT TGLLFDY 78 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83 ADI-30306 GSI S SS L YYWG 72 SIYYSGST F YNPS F KS 75 AREAGR GT TGLLFDY 78 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83 ADI-30311 GSI A SS V YYWG 73 SIYYSGST W YNPS L KS 76 AREAGR TG TGLLFDY 79 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83 Consensus Sequence GSIX ₁ SSX ₂ YYWG
(Where X ₁ is selected from A, S and residues that provide their conservative replacement, and X ₂ is selected from S, L, V and residues that provide their conservative replacement) 74 SIYYSGTX ₁ YNPSX ₂ KS
(where X ₁ is selected from Y, F, W and residues that provide their conservative replacement, and X ₂ is selected from L, F and residues that provide their conservative replacement) 77 AREAGRX ₁ X ₂ TGLLFDY
(Where X ₁ is selected from G, T and residues that provide their conservative replacement, and X ₂ is selected from T, G and residues that provide their conservative replacement) 80 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83

Table 2. CDR sequences of exemplary antibodies of the present invention

Antibody HCDR1-AbM SEQ HCDR2-Kabat SEQ HCDR3-Kabat SEQ LCDR1-Kabat SEQ LCDR2-Kabat SEQ LCDR3-Kabat SEQ ADI-27238 GYTFTSYYMS 178 IINPSGGSTSYAQKFQG 6 ARYPSSWPYGMDV 182 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30263 GYTFLSYYMN 179 IIDPSGGRTSYAQKFQG 7 ARYPSSWPYGTDV 183 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30267 GYTFRSYYMS 180 IIDPSGGRTSFAQKFQG eight ARYPESWPYGMDV 184 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-30268 GYTFGSYYMG 181 IIDPSGGRTSYARKFQG nine ARTPSSWPYGMDV 185 RASQSINSYLN 16 AASSLQS 17 QQSLTTPFT eighteen ADI-27243 GGSISSSRYYWG 186 SIYYSGSTYYNPSLKS 22 DGLYHTPEYFQH 188 RASQSVSSYLA 26 DASNRAT 27 QQRFAFPYT 28 ADI-30302 GGSIGSSQYYWG 187 SIYRSGGTYYNPSLKS 23 DGLYHTPEYFQH 188 RASQSVSSYLA 26 DASNRAT 27 QQRFAFPYT 28 ADI-30336 GGSISSSRYYWG 186 SIYYSGSTYYNPSLKS 22 DGLYHTPEYFQH 188 RASQSVSSYLA 26 DASNRAT 27 QQRFAHPYT 29 ADI-27278 GFTFSSYSMN 189 YISGSSSTIYYADSVKG 34 HRIADSPSRAFDI 191 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-30293 GFTFSSYSMN 189 YI-SSSGTINYADSVKG 35 HRIADSPSRAFDI 191 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-30296 GFTIGGYSMN 190 YI-SSSSTIHYADSVKG 36 HRIGRSPSRAFDI 192 KSSQSVLFSSNNKNYLA 41 WASTRES 42 QQSYFFPT 43 ADI-27291 GFTFSSYAMS 193 AISGSGGSTYYADSVKG 48 DPGTDSSGYYYIWRY 196 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30283 GFTFSPYGMS 194 SISGSGGRTYYADSVKG 49 DPGTDSSGYYYIWRY 196 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30286 GFTFGPYGMS 195 AISGSGASTWYADSVKG fifty DPGTHYSGYYYIWRY 197 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-30288 GFTFGPYGMS 195 AISGSGASTWHADSVKG 51 DPGTDSTGYYYIWRY 198 RASQSVSSNLA 57 SASTRAT 58 QQYVPHPPFT 59 ADI-27297 GYTFTSYYMH 199 IINPSGGSTSYAQKFQG 6 DHDIAAAGRLADY 201 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-30272 GYTFTEYYMH 200 IISPSAGSTSYAQKFQG 63 DHDIAAAGRLADY 201 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-30278 GYTFTEYYMH 200 IISPSAGSTKYAQKFQG 64 DDHIRLAGRLADY 202 RASQGISSWLA 69 AASSLQS 17 QQAVILPIT 70 ADI-27301 GGSISSSSYWG 203 SIYYSGSTYYNPSLKS 22 EAGRGTTGLLFDY 206 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83 ADI-30306 GGSISSSLYYWG 204 SIYYSGTFYNPSFKS 75 EAGRGTTGLLFDY 206 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83 ADI-30311 GGSIASSVYYWG 205 SIYYSGSTWYNPSLKS 76 EAGRTGTGLLFDY 207 RASQSISSWLA 81 KASSLES 82 QQYGILPRT 83

Table 3. VH and VL sequences of variable regions of exemplary antibodies of the present invention

VH sequences of exemplary antibodies Antibody vh SEQID NO: ADI-27238 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMSWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARARYPSSWPYGMDVWGQGTTVTVSS 84 ADI-30263 QVQLVQSGAEVKKPGASVKVSCKASGYTFLSYYMNWVRQAPGQGLEWMGIIDPSGGRTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARARYPSSWPYGTDVWGQGTTVTVSS 85 ADI-30267 QVQLVQSGAEVKKPGASVKVSCKASGYTFRSYYMSWVRQAPGQGLEWMGIIDPSGGRTSFAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARARYPESWPYGMDVWGQGTTVTVSS 86 ADI-30268 QVQLVQSGAEVKKPGASVKVSCKASGYTFGSYYMGWVRQAPGQGLEWMGIIDPSGGRTSYARKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARARTPSSWPYGMDVWGQGTTVTVSS 87 ADI-27243 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSRYYWGWIRQPPGKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARDGLYHTPEYFQHWGQGTLVTVSS 88 ADI-30302 QLQLQESGPGLVKPSETLSLTCTVSGGSIGSSQYYWGWIRQPPGKGLEWIGSIYRSGGTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARDGLYHTPEYFQHWGQGTLVTVSS 89 ADI-30336 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSRYYWGWIRQPPGKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARDGLYHTPEYFQHWGQGTLVTVSS 90 ADI-27278 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSYISGSSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARHRIADSPSRAFDIWGQGTMVTVSS 91 ADI-30293 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSYISSSGTINYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARHRIADSPSRAFDIWGQGTMVTVSS 92 ADI-30296 EVQLVESGGGLVQPGGSLRLSCAASGFTIGGYSMNWVRQAPGKGLEWVSYISSSSTIHYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARHRIGRSPSRAFDIWGQGTMVTVSS 93 ADI-27291 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPGTDSSGYYYIWRYWGQGTLVTVSS 94 ADI-30283 EVQLLESGGGLVQPGGSLRLSCAASGFTFSPYGMSWVRQAPGKGLEWVSSISGSGGRTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPGTDSSGYYYIWRYWGQGTLVTVSS 95 ADI-30286 EVQLLESGGGLVQPGGSLRLSCAASGFTFGPYGMSWVRQAPGKGLEWVSAISGSGASTWYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPGTHYSGYYYIWRYWGQGTLVTVSS 96 ADI-30288 EVQLLESGGGLVQPGGSLRLSCAASGFTFGPYGMSWVRQAPGKGLEWVSAISGSGASTWHADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPGTDSTGYYYIWRYWGQGTLVTVSS 97 ADI-27297 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDHDIAAAGRLADYWGQGTLVTVSS 98 ADI-30272 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEYYMHWVRQAPGQGLEWMGIISPSAGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDHDIAAAGRLADYWGQGTLVTVSS 99 ADI-30278 QVQLVQSGAEVKKPGASVKVSCKASGYTFTEYYMHWVRQAPGQGLEWMGIISPSAGSTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDHDIRLAGRLADYWGQGTLVTVSS 100 ADI-27301 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPPGKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREAGRGTTGLLFDYWGQGTLVTVSS 101 ADI-30306 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSLYYWGWIRQPPGKGLEWIGSIYYSGSTFYNPSFKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREAGRGTTGLLFDYWGQGTLVTVSS 102 ADI-30311 QLQLQESGPGLVKPSETLSLTCTVSGGSIASSVYYWGWIRQPPGKGLEWIGSIYYSGSTWYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREAGRTGTGLLFDYWGQGTLVTVSS 103 Exemplary Antibody VL Sequences Antibody VL SEQID NO: ADI-27238 DIQMTQSPSSLSASVGDRVTITCRASQSINSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSLLTPFTFGGGTKVEIK 104 ADI-30263 DIQMTQSPSSLSASVGDRVTITCRASQSINSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSLLTPFTFGGGTKVEIK 104 ADI-30267 DIQMTQSPSSLSASVGDRVTITCRASQSINSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSLLTPFTFGGGTKVEIK 104 ADI-30268 DIQMTQSPSSLSASVGDRVTITCRASQSINSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSLLTPFTFGGGTKVEIK 104 ADI-27243 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFAFPYTFGGGTKVEIK 105 ADI-30302 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFAFPYTFGGGTKVEIK 105 ADI-30336 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRFAHPYTFGGGTKVEIK 106 ADI-27278 DIVMTQSPDSLAVSLGERATINCKSSQSVLFSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSYFFPTFGGGTKVEIK 107 ADI-30293 DIVMTQSPDSLAVSLGERATINCKSSQSVLFSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSYFFPTFGGGTKVEIK 107 ADI-30296 DIVMTQSPDSLAVSLGERATINCKSSQSVLFSSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSYFFPTFGGGTKVEIK 107 ADI-27291 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYVPHPPFTFGGGTKVEIK 108 ADI-30283 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYVPHPPFTFGGGTKVEIK 108 ADI-30286 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYVPHPPFTFGGGTKVEIK 108 ADI-30288 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYVPHPPFTFGGGTKVEIK 108 ADI-27297 DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLISAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAVILPITFGGGTKVEIK 109 ADI-30272 DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLISAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAVILPITFGGGTKVEIK 109 ADI-30278 DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLISAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAVILPITFGGGTKVEIK 109 ADI-27301 DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYGILPRTFGGGTKVEIK 110 ADI-30306 DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYGILPRTFGGGTKVEIK 110 ADI-30311 DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYGILPRTFGGGTKVEIK 110

Table 4. SEQ ID numbers of related sequences of all antibodies that are the subject of the present invention

Antibody HCDR1 HCDR2 HDDR3 vh HC VH DNA LCDR1 LCDR2 LCDR3 VL LC VL DNA ADI-27238 one 6 eleven 84 111 150 16 17 eighteen 104 137 170 ADI-30263 2 7 12 85 112 151 16 17 eighteen 104 137 170 ADI-30267 3 eight 13 86 113 152 16 17 eighteen 104 137 170 ADI-30268 4 nine fourteen 87 114 153 16 17 eighteen 104 137 170 Consensus Sequence five 10 fifteen ADI-27243 nineteen 22 25 88 115 154 26 27 28 105 138 171 ADI-30302 twenty 23 25 89 116 155 26 27 28 105 138 171 ADI-30336 nineteen 22 25 90 117 156 26 27 29 106 139 172 Consensus Sequence 21 24 25 thirty ADI-27278 31 34 38 91 118 157 41 42 43 107 140 173 ADI-30293 31 35 38 92 119 158 41 42 43 107 140 173 ADI-30296 32 36 39 93 120 159 41 42 43 107 140 173 Consensus Sequence 33 37 40 ADI-27291 44 48 53 94 121 160 57 58 59 108 141 174 ADI-30283 45 49 53 95 122 161 57 58 59 108 141 174 ADI-30286 46 fifty 54 96 123 162 57 58 59 108 141 174 ADI-30288 46 51 55 97 124 163 57 58 59 108 141 174 Consensus Sequence 47 52 56 ADI-27297 60 6 66 98 125 164 69 17 70 109 142 175 ADI-30272 61 63 66 99 126 165 69 17 70 109 142 175 ADI-30278 61 64 67 100 127 166 69 17 70 109 142 175 Consensus Sequence 62 65 68 ADI-27301 71 22 78 101 128 167 81 82 83 110 143 176 ADI-30306 72 75 78 102 129 168 81 82 83 110 143 176 ADI-30311 73 76 79 103 130 169 81 82 83 110 143 176 Consensus Sequence 74 77 80

Note. HC was formed by fusion of the VH sequence with the IgG4-PAA portion of the sequence shown in SEQ ID NO: 177, and LC was formed by fusion of the VL sequence with the Cκ chain of the sequence shown in SEQ ID NO: 209.

20 exemplary antibodies (ADI-27238, ADI-30263, ADI-30267, ADI-30268, ADI-27243, ADI-30302, ADI-30336, ADI-27278, ADI-30293, ADI-30296, ADI-27291, ADI- 30283, ADI-30286, ADI-30288, ADI-27297, ADI-30272, ADI-30278, ADI-27301, ADI-30306 and ADI-30311) included in the examples of the present invention described below, CDR amino acid sequences, variable the light and heavy chain regions, light and heavy chains of these antibodies, as well as the corresponding nucleotide sequences, are shown in Tables 1-3 and in the sequence listing of the present application. The corresponding sequence numbers are shown in Table 4 .

Examples

Example 1 Obtaining Antibodies

Screening of fully humanized anti-TIGIT antibodies using yeast display

Yeast antibody libraries (Adimab) were amplified by methods known to those skilled in the art (described in WO 2009036379, WO 2010105256 and WO 2012009568), with a diversity of each library of 1×10 ⁹ . Briefly, in the first two rounds of screening, cells were sorted on magnetic beads using a MACS system manufactured by Miltenyi. First, yeast cells (approximately 1x10 ¹⁰ cells/library) from the libraries were incubated in FACS buffer (phosphate buffer containing 0.1% bovine serum albumin and 100 nM human TIGIT antigens labeled with biotin (Acro Biosystems, TIT-H52H3)) in for 15 min at room temperature. Cells were washed once with 50 ml of pre-chilled FACS buffer and resuspended in 40 ml of the same buffer, after which 500 μl of streptavidin microbeads (Miltenyi LS) were added and incubated at 4°C for 15 min. The mixture was centrifuged at 1000 rpm for 5 minutes. After separation of the supernatant, the cells were resuspended in 5 ml of FACS buffer. The resulting cell suspension was applied to a Miltenyi LS column. After loading, the column was washed three times with FACS buffer, each time with 3 ml. The Miltenyi LS column was removed from the magnetic field and eluted with 5 ml of growth medium. The eluted yeast cells were collected and incubated overnight at 37°C.

The next round of sorting was performed using a flow cytometer; while approximately 1×10 ⁸ yeast cells selected using the MACS system were washed three times with FACS buffer and incubated together with TIGIT antigens labeled with low concentration biotin (100 ^-1 nM) at room temperature. The supernatant was discarded. Cells were washed twice with FACS buffer and mixed with LC-FITC (FITC labeled anti-human immunoglobulin kappa light chain antibody, Southern Biotech) (1:100 dilution) and SA-633 (Streptavidin-633, Molecular Probes) (1:500 dilution) or SA-PE (streptavidin-PE, Sigma) (1:50 dilution), after which the mixture was incubated at 4°C for 15 min. Cells were eluted twice with pre-chilled FACS buffer, resuspended in 0.4 ml of buffer, and transferred to a separation tube with a filter. Cells were sorted using a FACS ARIA instrument (BD Biosciences).

Yeast cells expressing anti-TIGIT antibodies obtained from the screening were induced by shaking at 30° C. for 48 hours to express anti-TIGIT antibodies. After induction, yeast cells were separated by centrifugation at 1300 rpm for 10 min and the supernatant was collected. Before harvesting, anti-TIGIT antibodies contained in the supernatant were purified with Protein A and eluted using acetate buffer pH 2.0. The purity of the antibodies was over 95%. Antibodies were digested with papain and purified on KappaSelect resin (GE Healthcare) to obtain the corresponding Fab fragments. This screening resulted in anti-TIGIT antibodies (ADI-27301, ADI-27238, ADI-27278, ADI-27243, ADI-27297 and ADI-27291).

Affinity optimization of antibodies to human TIGIT

To generate higher affinity anti-human TIGIT antibodies (ADI-27301, ADI-27238, ADI-27278, ADI-27243, ADI-27297 and ADI-27291) were optimized by the following methods.

VHmut screening

This method was used to introduce mutations in the region of heavy chains of antibodies using standard mismatched PCR. The probability of base mismatch during PCR was increased to approximately 0.01 bp. by adding 1 μM dPTP and 8-oxo-dGTP base analogs with high mutation potential.

The resulting mismatched PCR products were introduced into a vector containing the heavy chain constant region by homologous recombination. This method generated a secondary library with a capacity of 1×10 ⁷ under stringent screening conditions, including titers to TIGIT antigens, competition with unlabeled antigen, and competition with parental (original) antibodies. Three rounds of screening have been successfully completed using FACS.

Screening for CDRH1/CDRH2

The CDRH3 genes of the VHmut progeny antibodies were used to construct a 1×10 ⁸ CDRH1/CDRH2 gene pool and performed 3 rounds of gene screening. MACS was used in the first round of screening, and FACS was used in the second and third rounds. The antigen-antibody conjugates were subjected to rigorous ("pressure") screening to select antibodies with maximum affinity.

As a result of the above affinity maturation process of 6 parental antibodies, 14 monoclonal antibodies to human TIGIT with improved affinity were obtained: ADI-30263/ADI-30267/ADI-30268 (from ADI-27238), ADI-30302/ADI-30336 (from ADI- 27243), ADI-30293/ADI-30296 (from ADI-27278), ADI-30283/ADI-30286/ADI-30288 (from ADI-27291), ADI-30272/ADI-30278 (from ADI-27297), and ADI -30306/ADI-30311 (from ADI-27301).

Yeast cells expressing anti-TIGIT antibodies obtained from the screening were induced by shaking at 30° C. for 48 hours to express anti-TIGIT antibodies. After induction, yeast cells were separated by centrifugation at 1300 rpm for 10 min and the supernatant was collected. Before harvesting, anti-TIGIT antibodies contained in the supernatant were purified with Protein A and eluted using acetate buffer pH 2.0. The purity of the antibodies was over 95%. Antibodies were digested with papain and purified on KappaSelect resin (GE Healthcare) to obtain the corresponding Fab fragments.

Expression and purification of antibodies

Sequence information and sequence numbers of antibodies to human TIGIT, which are the subject of the present invention, are presented in Tables 1-9; while both parent antibodies and daughter antibodies after affinity maturation were expressed and purified in yeast by the method described above.

The following reference antibodies used in the examples were expressed and purified in HEK293 cells: 22G2, an anti-human TIGIT antibody from BMS, which was transiently expressed in HEK293 cells with identical light and heavy chain variable region sequences to those of the "22G2" antibody described in patent application no. WO 2016/106302A1; 31C6, an anti-human TIGIT antibody from Merck, which was transiently expressed in HEK293 cells with identical light and heavy chain variable region sequences to those of the "31C6" antibody described in Patent Application No. WO 2016/028656A1; 10A7 and 1F4, anti-human TIGIT antibodies from Genentech, which were transiently expressed in HEK293 cells, with the light and heavy chain variable region sequences identical to those of the "10A7" and "1F4" antibodies in Patent Application No. WO2015009856A2, respectively. The constant regions of 4 reference antibodies corresponded to wild-type IgG4 sequences.

The pTT5 vector was used to transiently express the antibody in HEK293 cells. The antibody heavy and light chains were first cloned into separate pTT5 vectors. The pTT5 vectors carrying the heavy and light chains of the antibody molecule were transferred into HEK293 cells by chemical transfection. Cultured HEK293 cells were transit transfected with PEI chemical transfection reagent (purchased from Polysciences) according to the scheme provided by the manufacturer. Plasmid DNA and transfection reagent were prepared in a laminar flow cabinet, after which F17 medium (Gibco) was transferred into two 50 ml centrifuge tubes (volume was 1/5 of the volume required for transfection). Filtered plasmids (130 µg/100 ml) were added to one tube and filtered PEI (1 g/L, Polysciences) (plasmid:PEI mass ratio = 1:3) was added to another tube. The contents of each tube were thoroughly mixed for 5 min, then both mixtures were combined and gently mixed 20 times, after which the resulting mixture was left alone for 15–30 min (no more than 30 min). The DNA/PEI mixture was carefully added to the HEK293 cells and mixed thoroughly. Cells were cultured at 37°C, 8% CO ₂ for 7 days, adding fresh medium every 48 hours. Seven days later, or when the percentage of viable cells in continuous culture reached ≤60%, the mixture was centrifuged at 13,000 rpm for 20 minutes. The supernatant was collected and the antibodies were purified on Protein A, resulting in antibodies with a purity greater than 95%.

In addition, intact antibodies (ADI-30268, ADI-30278, ADI-30286, ADI-30288, ADI-30293, ADI-30306 and ADI-30336) were expressed and purified in CHO cells.

Expression and purification in CHO cells: CHO cell lines expressing antibodies were generated using the HD-BIOP1 line (GS Null CHO-K1) from Horizon according to the manufacturer's instructions. The heavy and light chain DNA sequences of the antibody molecule were first inserted into ATUM pD2531 plasmids. The constructed plasmids were then transferred into CHO cells by electrotransfection, and antibody production was detected by the ForteBio method, which made it possible to determine the efficiency of transfection 24 h after it. The transfected cells were subjected to rigorous ("pressure") screening to obtain a pool of cells with a high level of antibody expression. This cell pool was then amplified to express large amounts of antibodies. The supernatant was collected from the cells and the antibodies were purified using Protein A and gel filtration, resulting in antibodies with a purity of more than 95%.

Example 2 Antibody Affinity Assay

Equilibrium dissociation constants (K _D ) characterizing the binding of 20 exemplary antibodies of the present invention to human TIGIT were measured by biological optical interferometry (ForteBio).

ForteBio affinity analysis was performed as previously described (Estep, P, et al. , High throughput solution based measurement of antibody-antigen affinity and epitope binning. MAbs , 2013.5(2): p. 270-8).

Measurement of monovalent binding affinity of a candidate Fab fragment to TIGIT-FC: the sensor was equilibrated offline in assay buffer for 20 min, followed by online detection for 120 s to establish a baseline. Human TIGIT-FC was applied to the AHQ sensor (ForteBio) for affinity analysis according to the ForteBio method. The probe coated with antigens was placed in a solution containing 100 nM Fab fragment until a plateau was reached, after which it was transferred for at least 2 min into the assay buffer in which dissociation occurred to measure the dissociation rate. Kinetic analysis was performed using a 1:1 binding model.

Measurement of monovalent binding affinity of intact candidate antibodies to human (ACRO, TIT-H52H3), mouse (ACRO, TIT-M52E6) and monkey (ACRO, TIT-C5223) TIGIT-his: the sensor was equilibrated offline in assay buffer for 20 min , followed by online detection for 120 s to establish a baseline. Purified antibodies were applied to the AHQ sensor (ForteBio) to a thickness of 1 nanometer for affinity analysis according to the ForteBio method. The antibody-coated probe was placed in a solution containing 100 nM TIGIT-his antigens until a plateau was reached, after which it was transferred for at least 2 min into assay buffer in which dissociation occurred to measure the dissociation rate. Kinetic analysis was performed using a 1:1 binding model.

In experiments using the assays described above, K _D values were determined for 20 candidate antibodies (Fab) expressed in yeast; these values are shown in Table 5.

Table 5. K _D values for monovalent binding of 20 candidate antibodies (Fab) expressed in yeast to human TIGIT-FC

Parental Subsidiaries ADI-27238 ADI-30263 ADI-30267 ADI-30268 1.38E-08 7.95E-10 1.66E-09 5.71E-10 ADI-27243 ADI-30302 ADI-30336 9.47E-09 3.29E-10 5.66E-10 ADI-27278 ADI-30293 ADI-30296 1.09E-08 3.49E-09 2.43E-09 ADI-27291 ADI-30283 ADI-30286 ADI-30288 6.48E-09 5.53E-10 4.36E-10 5.47E-10 ADI-27297 ADI-30272 ADI-30278 3.21E-08 1.19E-09 2.43E-10 ADI-27301 ADI-30306 ADI-30311 8.01E-09 7.29E-10 8.07E-10

In experiments using the assays described above, K _D values were determined for monovalent binding of 7 intact candidate antibodies expressed by CHO cells to human, mouse, and monkey TIGIT; these values are shown in Table 6.

Table 6. K _D values for monovalent binding of 7 candidate antibodies expressed by CHO cells and reference antibodies to human, mouse and monkey TIGIT

No. K _D (human) K _D (monkey) K _D (mouse) ADI-30268 3.29E-09 6.23E-08 4.64E-08 ADI-30278 1.75E-09 4.53E-09 N, B ADI-30286 1.76E-09 1.06E-08 1.34E-07 ADI-30288 1.31E-09 7.19E-09 5.70E-09 ADI-30293 7.89E-10 3.49E-08 1.30E-08 ADI-30306 2.70E-09 3.44E-09 N, B ADI-30336 1.46E-09 5.56E-10 N, B 22G2 (BMS) 3.50E-09 4.20E-09 N, B 31C6 (MSD) 2.34E-09 4.72E-10 N, B

From the tables above, it can be seen that: (1) after affinity maturation, each daughter antibody has a significantly higher binding affinity for human TIGIT than the parent antibody; (2) after affinity maturation, the binding affinity of the 7 selected antibodies expressed in CHO cells to human TIGIT was about the same or higher than that of the reference antibodies; all 7 candidate antibodies can cross-react with monkey TIGIT; while 4 (ADI-30268/ADI-30286/ADI-30288/ADI-30293) out of 7 candidate antibodies can cross-react with mouse TIGIT.

Example 3 Binding of anti-TIGIT antibodies to human TIGIT expressed on cell surfaces

The ability of the 20 above exemplary antibodies of the present invention to bind to human TIGIT expressed on the surface of CHO cells was measured by flow cytometry. The binding capacity of various antibodies was determined by comparing their binding curves with human TIGIT expressed on the surface of CHO cells.

In particular, yeast cells were used to express parent and daughter antibodies after affinity maturation. After collecting antibodies (with a purity of about 70%), experiments were performed to bind these antibodies to human TIGIT expressed on the surface of CHO cells. Experiments were performed as follows: (1) CHOS cells were transfected with pCHO1.0-engineered human TIGIT-bearing plasmids, and then screened with methotrexate and puromycin to select a pool of stable cells. (2) Candidate antibodies at various concentrations were incubated with a pool of stable cells at 4°C for 30 min. After three washes with phosphate buffered saline (PBS), the cells were incubated with phycoerythrin (PE) labeled mouse second antibodies to human immunoglobulins for 30 min at 4°C. After three additional washes with PBS, the cells were resuspended in 100 µl of PBS. (3) Using flow cytometry, the median fluorescence values for cells in two channels were determined. EC ₅₀ values and maximum values for different curves were compared using graphs constructed in the coordinates "decimal logarithm of antibody concentration" (along the abscissa axis) - "fluorescence median for two channels" (along the ordinate axis). The results are shown in the following table and in FIG. 1 (EC ₅₀ : nM).

Antibody IgG1 10A7 1F4 22G2 31C6 _EC50 ~1.388e+007 0.2690 0.7096 0.5106 0.4358 Antibody ADI-27238 ADI-30267 ADI-30268 ADI-27243 ADI-30302 ADI-30336 _EC50 0.7512 0.7402 0.3293 0.5873 0.3262 0.6291 Antibody ADI-27278 ADI-30293 ADI-30296 ADI-27291 ADI-30286 ADI-30288 _EC50 0.9682 0.9003 0.3714 0.8522 0.4459 0.4755 Antibody ADI-27297 ADI-30272 ADI-30278 ADI-27301 ADI-30306 ADI-30311 _EC50 0.4958 0.6098 0.4086 0.8990 0.7634 0.7566

From the above experimental results, the following conclusions can be drawn: (1) after affinity maturation, each daughter antibody has a higher ability to bind to human TIGIT expressed on the surface of CHO cells than the parent antibody (as indicated by a higher plateau in the binding curve); (2) the ability of progeny antibodies, after affinity maturation, to bind to human TIGIT expressed on the surface of CHO cells is higher than for the two antibodies from Genentech (10A7 and 1F4), and is comparable to that of the 22G2 antibody from BMS and the 31C6 antibody from the company Merck.

In addition, 7 antibodies selected after affinity maturation were expressed and purified using the CHO system to obtain high purity antibodies. An experiment was conducted to bind high purity antibodies to human TIGIT expressed on the surface of CHO cells. In the experiments performed as described above, antibodies ADI-30268, ADI-30336, ADI-30293, ADI-30286, ADI-30288, ADI-30278, and ADI-30306 bound to human TIGIT overexpressed on the surface of CHO cells, with values EC ₅₀ 0.1392 nM, 0.1300 nM, 0.2288 nM, 0.1758 nM, 0.1860 nM, 0.1321 nM, and 0.1999 nM, respectively, with their ability to bind to human TIGIT overexpressed on cell surfaces CHO was higher than that of reference antibodies 22G2 and 1F4 (EC ₅₀ values are 0.7249 nM and 0.6073 nM, respectively) and comparable to that of reference antibody 31C6 (EC ₅₀ value is 0.1846 nM). Although the EC ₅₀ value for the reference antibody 10A7 (0.1949 nM) is comparable to those for the 7 candidate antibodies, it can be seen from the binding curve of this reference antibody that the plateau in this curve is lower than the plateau in the curves of the 7 candidate antibodies. As a rule, this indicates a more rapid dissociation of the antigen-antibody complex (see FIG. 2). Therefore, the binding capacity of 7 candidate antibodies to human TIGIT expressed on the surface of CHO cells was found to be higher than that of the 10A7 antibody.

Example 4 Anti-TIGIT Antibody Blocks TIGIT Binding to Its CD155 Ligand

The ability of candidate antibodies to block ligands was determined by flow cytometry. This determination was carried out as follows: to the resulting pool of CHOS cells stably overexpressing human TIGIT, a concentration of 50 nM human CD155 protein (ACRO, TIT-H5253-1MG) with a mouse IgG2a Fc fragment, as well as antibodies at various concentrations, was added, after which cells were incubated for 30 min at 4°C. After three washes with PBS, the CD155 ligand molecules remaining on the cells were stained with goat anti-murine immunoglobulin Fc (at 1%) and a second antibody labeled with allophycocyanin (APC) (Biolegend, 405308). After three more washes with PBS, the median fluorescence in the corresponding channel (C6, channel 4) was measured using a flow cytometer. For each antibody, the median fluorescence (y-axis) was plotted against the decimal logarithm of antibody concentration in nM (abscissa). The ability of various antibodies to block the binding of TIGIT to CD155 was assessed by IC ₅₀ values, which were determined from the generated graphs.

In experiments to evaluate the ability of various candidate antibodies before and after affinity maturation to block the binding of TIGIT to CD155, IgG1 was used as a negative control, and antibodies 10A7, 22G2 and 31C6 were used as positive controls. RICE. 3 and the table below demonstrate the ability of antibodies expressed in yeast before and after affinity maturation to block TIGIT binding to CD155 (IC ₅₀ : nM).

Antibody IgG1 10A7 22G2 31C6 IC50 ~0.03904 0.7408 0.1786 0.1536 Antibody ADI-27238 ADI-30267 ADI-30268 ADI-27243 ADI-30302 ADI-30336 IC50 0.4181 0.2775 0.2209 0.8188 0.2808 0.4093 Antibody ADI-27278 ADI-30293 ADI-30296 ADI-27291 ADI-30286 ADI-30288 IC50 0.5859 0.8586 0.2781 1.614 0.2965 0.6677 Antibody ADI-27297 ADI-30272 ADI-30278 ADI-27301 ADI-30306 ADI-30311 IC50 0.6029 0.4036 0.2175 0.6411 0.5281 0.5313

From the results of the experiments, it can be concluded that most of the daughter antibodies after affinity maturation had a higher ability to block the binding of TIGIT to CD155 than their parent antibodies, were comparable in blocking ability to the reference antibodies 22G2 and 31C6, and surpassed the reference antibody 10A7.

The ability to block TIGIT binding to CD155 was also determined for 7 high purity antibodies expressed in CHO cells. In the experiments performed as described above, IgG4 was used as a negative control, and antibodies 22G2, 31C6 and 10A7 were used as positive controls. From the results of the experiments, we can conclude that the IC ₅₀ values for antibodies ADI-30268, ADI-30336 and ADI-30278 are 0.1402 nM, 0.1219 nM and 0.1164 nM, respectively, that is, lower than those for the reference antibodies 22G2 (0.6069 nM), 31C6 (0.1659 nM) and 10A7 (0.3252 nM), which means that these three antibodies have a higher ability to block ligand binding (CD155) than the reference antibodies. Four other candidate antibodies (ADI-30293, ADI-30286, ADI-30288, and ADI-30306) have IC ₅₀ values of 0.2510 nM, 0.1893 nM, 0.2238 nM, and 0.2420 nM, respectively; all of these values are lower than the values for the reference antibodies 22G2 and 10A7, but higher than those for the reference antibody 31C6, which implies that these 4 antibodies have a higher ability to block ligand binding (CD155) than the reference antibodies 22G2 and 10A7, but lower, than the reference antibody 31C6. The results are shown in the following table and in FIG. 4 (IC ₅₀ : nM).

Antibody IgG1 22G2 31C6 10A7 _IC50 ~1,152 0.6069 0.1659 0.3252 Antibody ADI-30268 ADI-30336 ADI-30293 ADI-30286 _IC50 0.1402 0.1219 0.2510 0.1893 Antibody ADI-30288 ADI-30278 ADI-30306 _IC50 0.2238 0.1164 0.2420

Example 5. Analysis of the biological activity of candidate antibodies underlying their mechanism of action (MD)

All anti-TIGIT antibodies, by blocking the binding of TIGIT to CD155, are able to attenuate the inhibitory effect on the downstream IL-2-dependent signaling pathway. In this example, the cell line for the analysis of the biological activity underlying MD (J2201) from Promega was used to detect the expression of fluorescent reporter protein genes, reflecting the activation of the IL-2-dependent signaling pathway, and thereby to determine the ability of antibodies to suppress binding TIGIT with CD155. The analysis was carried out according to the manufacturer's instructions, Promega. The analysis procedure was as follows: Jurkat cells stably expressing human TIGIT and luciferase reporter protein genes under the control of IL-2 promoters, CHO-K1 cells stably expressing human CD155 and T-lymphocyte activation elements, and antibodies to human TIGIT at various concentrations together. incubated in a CO ₂ incubator at 37°C for 6 h, after which the fluorescence intensity was measured.

The results of the experiments showed that the daughter antibodies after affinity maturation (directly expressed by yeast, with a purity of approximately 70%) had a significantly higher in vitro biological activity than their parent antibodies, and in most cases reached a level of biological activity comparable to that of the two reference antibodies 22G2 and 31C6. The results are shown in the following table and in FIG. 5 (EC ₅₀ : nM).

Antibody IgG1 10A7 22G2 31C6 _EC50 ~272.8 ~77.84 2.113 0.8383 Antibody ADI-27238 ADI-30267 ADI-30268 _EC50 68.70 2.064 1.224 Antibody ADI-27243 ADI-30302 ADI-30336 _EC50 297.1 0.9831 3.334 Antibody ADI-27278 ADI-30293 ADI-30296 _EC50 ~1.405e+008 1.559 1.088 Antibody ADI-27297 ADI-30272 ADI-30278 _EC50 ~25059 1.489 0.5228 Antibody ADI-27301 ADI-30306 ADI-30311 _EC50 ~1.085e+012 1.645 1.824

Analysis of the biological activity underlying MD was also carried out for 7 highly purified antibodies expressed by CHO cells. In the assay described above, IgG4 was used as a negative control and the 31C6 antibody was used as a positive control. From the results of the experiments, we can conclude that the candidate antibody ADI-30278 is characterized by an EC ₅₀ value of 0.1856 nM, which is lower than the corresponding values for other candidate antibodies and for the reference antibody 31C6 (0.3424 nM), which indicates higher biological activity in vitro ; antibodies ADI-30268, ADI-30286, ADI-30288 and ADI-30306 are characterized by EC ₅₀ values of 0.5065 nM, 0.3963 nM, 0.3844 nM and 0.3984 nM, respectively, that is, they have approximately the same biological activity, as the reference antibody 31C6; antibodies ADI-30336 and ADI-30293 have EC ₅₀ values of 1.274 nM and 1.624 nM, respectively, indicating lower biological activity. The results are shown in the following table and in FIG. 6 (EC ₅₀ : nM).

Antibody IgG4 31C6 ADI-30268 EC50 0.0 0.3424 0.5065 Antibody ADI-30336 ADI-30293 ADI-30286 EC50 1.274 1.624 0.3963 Antibody ADI-30288 ADI-30278 ADI-30306 EC50 0.3844 0.1856 0.3984

Example 6 Efficiency Study in vivo (for animals)

This study investigated the antitumor activity of anti-TIGIT antibodies in an experimental tumor model in MC38 mice with a knock-in human TIGIT gene.

We studied the pharmacodynamics of candidate antibodies ADI-30293 and ADI-30278, which were administered separately (at a dose of 10 mg/kg) to MC38 mice with an introduced human TIGIT receptor gene. Mouse MC38 colon cancer cells (OBiO Technology (Shanghai) Co., Ltd., HYC0116) were transplanted into female TIGIT transgenic mice (Biocytogen Beijing Co., Ltd.). Mice from each group were injected subcutaneously with antibodies on days 6, 10, 13 and 17 after transplantation. On days 6, 10, 13, 17, 21 and 25 after transplantation, the tumor volume was measured, after which the mice were euthanized. The results are shown in FIG. 7.

From the results shown in FIG. 7, it can be seen that the anti-TIGIT monoclonal antibodies ADI-30293 and ADI-30278 of the present invention, when used alone, suppressed tumor growth to a certain extent compared to Ig4, which served as a negative control. In terms of ability to suppress tumor growth, the two candidate antibodies, when used alone, were not statistically significantly different from the 31C6 antibody, which served as a positive control.

Additionally, we studied the pharmacodynamics of candidate antibodies ADI-30293 and ADI-30278, which were administered (at a dose of 10 + 1 mg/kg) in combination with an antibody to PD1 (antibody C, WO2017133540A1) to MC38 mice with an inserted (knock-in) TIGIT gene. person.

Mouse colon cancer cells MC38 (OBiO Technology (Shanghai) Co., Ltd., HYC0116) were cultured in DMEM. 1×10 ⁶ MC38 cells in 0.2 ml of DMEM basic suspension medium were transplanted onto the right flank of female transgenic mice with the introduced TIGIT gene (SHANGHAI MODEL ORGANISMS). Tumor volume and body weight were measured twice a week during the study. On the 8th day after tumor cell transplantation, the length and width of the tumor were measured with a caliper. Tumor volume was calculated using the following formula: width ² × length/2 (mm ³ ). Mice with an average tumor volume of approximately 71 mm ^{3 were} randomly divided into groups (6 mice each) and euthanized after reaching the endpoint for determining tumor volume or when mice lost more than 20% of body weight. On days 8, 12, 15, and 19 post-transplant, mice from each group were subcutaneously injected with human IgG (10 mg/kg), antibody C (1 mg/kg), and anti-TIGIT antibodies or their isotypes (10 mg/kg), respectively. The mean endpoint tumor volume was determined on day 29 post-cell transplantation, after which the mice were euthanized. The results are shown in FIG. eight.

From the results shown in FIG. 8, it can be seen that the anti-TIGIT antibody (ADI-30293) used in combination with the anti-PD1 antibody (antibody C) inhibited tumor growth more strongly than either antibody alone; the inhibitory effect of the combination of ADI-30293 antibody and anti-PD1 antibody is superior to that of the combination of 31C6 antibody and anti-PD1 antibody. Similarly, the effect of the combination of ADI-30278 antibody and anti-PD1 antibody was superior to that of the combination of 31C6 antibody and anti-PD1 antibody (antibody C).

In this study, there was no statistically significant change in the body weight of mice at day 29 after cell transplantation in any of the groups.

--->

SEQUENCE LIST

<110> Innovent Biologics (Suzhou) Co., Ltd.

<120> Antibody to TIGIT and its use

<130> PF190269PCT

<160> 209

<170> Patent revision 3.3

<210> 1

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 1 ADI-27238 HCDR1 KABAT,

numbered positions H27-35B

<400> 1

Tyr Thr Phe Thr Ser Tyr Tyr Met Ser

fifteen

<210> 2

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> ADI-30263 HCDR1 KABAT antibody sequence 2,

numbered positions H27-35B

<400> 2

Tyr Thr Phe Leu Ser Tyr Tyr Met Asn

fifteen

<210> 3

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 3 ADI-30267 HCDR1 KABAT,

numbered positions H27-35B

<400> 3

Tyr Thr Phe Arg Ser Tyr Tyr Met Ser

fifteen

<210> 4

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 4 ADI-30268 HCDR1 KABAT,

numbered positions H27-35B

<400> 4

Tyr Thr Phe Gly Ser Tyr Tyr Met Gly

fifteen

<210> 5

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 5 consensus sequence

<220>

<221> misc_feature

<222> (4)..(4)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from T, L, R, G, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(9)..(9)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, N, G, and its conservatively substituted residue

<400> 5

Tyr Thr Phe Xaa Ser Tyr Tyr Met Xaa

fifteen

<210> 6

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 6 ADI-27238, ADI-27297 HCDR2

KABAT, numbered positions H50-65

<400> 6

Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln

1 5 10 15

gly

<210> 7

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 7 ADI-30263 HCDR2 KABAT, H50-65

<400> 7

Ile Ile Asp Pro Ser Gly Gly Arg Thr Ser Tyr Ala Gln Lys Phe Gln

1 5 10 15

gly

<210> 8

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 8 ADI-30267 HCDR2 KABAT, H50-65

<400> 8

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

1 5 10 15

gly

<210> 9

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 9 ADI-30268 HCDR2 KABAT, H50-65

<400> 9

Ile Ile Asp Pro Ser Gly Gly Arg Thr Ser Tyr Ala Arg Lys Phe Gln

1 5 10 15

gly

<210> 10

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 10 consensus sequence

<220>

<221> misc_feature

<222> (3)..(3)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from N, D, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(8)..(8)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, R, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (11)..(11)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Y, F, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (13)..(13)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Q, R, and its conservatively substituted residue

<400> 10

Ile Ile Xaa Pro Ser Gly Gly Xaa Thr Ser Xaa Ala Xaa Lys Phe Gln

1 5 10 15

gly

<210> 11

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 11 ADI-27238 HCDR3 Kabat, H93-102

<400> 11

Ala Arg Ala Arg Tyr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val

1 5 10 15

<210> 12

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 12 ADI-30263 HCDR3 Kabat, H93-102

<400> 12

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

1 5 10 15

<210> 13

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 13 ADI-30267 HCDR3 Kabat, H93-102

<400> 13

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

1 5 10 15

<210> 14

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 14 ADI-30268 HCDR3 Kabat, H93-102

<400> 14

Ala Arg Ala Arg Thr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val

1 5 10 15

<210> 15

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 15, consensus sequence

<220>

<221> misc_feature

<222> (5)..(5)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Y, T, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (13)..(13)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from M, T, and its conservatively substituted residue

<400> 15

Ala Arg Ala Arg Xaa Pro Ser Ser Trp Pro Tyr Gly Xaa Asp Val

1 5 10 15

<210> 16

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 16 ADI-27238, ADI-30263, ADI-30267,

ADI-30268 LCDR1 Kabat, L24-34

<400> 16

Arg Ala Ser Gln Ser Ile Asn Ser Tyr Leu Asn

1 5 10

<210> 17

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 17 ADI-27238, ADI-30263, ADI-30267,

ADI-30268, ADI-27297, ADI-30272, ADI-30272 LCDR2 Kabat, L50-56

<400> 17

Ala Ala Ser Ser Leu Gln Ser

fifteen

<210> 18

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 18 ADI-27238, ADI-30263, ADI-30267,

ADI-30268 LCDR3 Kabat, L89-97

<400> 18

Gln Gln Ser Leu Leu Thr Pro Phe Thr

fifteen

<210> 19

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 19 ADI-27243, ADI-30336 HCDR1

KABAT, numbered positions H27-35B

<400> 19

Gly Ser Ile Ser Ser Ser Arg Tyr Tyr Trp Gly

1 5 10

<210> 20

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 20 ADI-30302 HCDR1 KABAT,

numbered positions H27-35B

<400> 20

Gly Ser Ile Gly Ser Ser Gln Tyr Tyr Trp Gly

1 5 10

<210> 21

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 21 sequence, consensus sequence

<220>

<221> misc_feature

<222> (4)..(4)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, G, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from R, Q, and its conservatively substituted residue

<400> 21

Gly Ser Ile Xaa Ser Ser Xaa Tyr Tyr Trp Gly

1 5 10

<210> 22

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 22 ADI-27243, ADI-30336, ADI-27301 HCDR2 KABAT, H50-65

<400> 22

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

1 5 10 15

<210> 23

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 23 ADI-30302 HCDR2 KABAT, H50-65

<400> 23

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

1 5 10 15

<210> 24

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 24, consensus sequence

<220>

<221> misc_feature

<222> (4)..(4)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Y, R, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, G, and its conservatively substituted residue

<400> 24

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

1 5 10 15

<210> 25

<211> 14

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 25 ADI-27243, ADI-30302, ADI-30336

HCDR3 Kabat, H93-102

<400> 25

Ala Arg Asp Gly Leu Tyr His Thr Pro Glu Tyr Phe Gln His

1 5 10

<210> 26

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 26 ADI-27243, ADI-30302, ADI-30336

LCDR1 Kabat, L24-34

<400> 26

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 27

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 27 ADI-27243, ADI-30302, ADI-30336

LCDR2 Kabat, L50-56

<400> 27

Asp Ala Ser Asn Arg Ala Thr

fifteen

<210> 28

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 28 ADI-27243, ADI-30302 LCDR3 Kabat,

L89-97

<400> 28

Gln Gln Arg Phe Ala Phe Pro Tyr Thr

fifteen

<210> 29

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 29 ADI-30336 LCDR3 Kabat, L89-97

<400> 29

Gln Gln Arg Phe Ala His Pro Tyr Thr

fifteen

<210> 30

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 30, consensus sequence

<220>

<221> misc_feature

<222>(6)..(6)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from F, H, and its conservatively substituted residue

<400> 30

Gln Gln Arg Phe Ala Xaa Pro Tyr Thr

fifteen

<210> 31

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 31 ADI-27278, ADI-30293 HCDR1

KABAT, numbered positions H27-35B

<400> 31

Phe Thr Phe Ser Ser Tyr Ser Met Asn

fifteen

<210> 32

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 32 ADI-30296 HCDR1 KABAT,

numbered positions H27-35B

<400> 32

Phe Thr Ile Gly Gly Tyr Ser Met Asn

fifteen

<210> 33

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 33, consensus sequence

<220>

<221> misc_feature

<222> (3)..(5)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 3 is selected from F, I and their conservatively substituted residues,

Xaa at position 4 is selected from S, G and their conservatively substituted residues,

and Xaa at position 5 is selected from S, G and their conservatively substituted residues

<400> 33

Phe Thr Xaa Xaa Xaa Tyr Ser Met Asn

fifteen

<210> 34

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 34 ADI-27278 HCDR2 KABAT, H50-65

<400> 34

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

1 5 10 15

gly

<210> 35

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 35 ADI-30293 HCDR2 KABAT, H50-65

<400> 35

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

1 5 10 15

<210> 36

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 36 ADI-30296 HCDR2 KABAT, H50-65

<400> 36

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

1 5 10 15

<210> 37

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 37, consensus sequence

<220>

<221> misc_feature

<222> (3)..(4)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 3 is selected from the deletion, S and its conservatively substituted residue,

and Xaa at position 4 is selected from G, S and their conservatively substituted residues

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, G, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (10)..(10)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Y, N, H, and its conservatively substituted residue

<400> 37

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

1 5 10 15

gly

<210> 38

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 38 ADI-27278, ADI-30293 HCDR3 Kabat, H93-102

<400> 38

Ala Arg His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile

1 5 10 15

<210> 39

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 39 ADI-30296 HCDR3 Kabat, H93-102

<400> 39

Ala Arg His Arg Ile Gly Arg Ser Pro Ser Arg Ala Phe Asp Ile

1 5 10 15

<210> 40

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 40 sequence, consensus sequence

<220>

<221> misc_feature

<222> (6)..(7)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 6 is selected from A, G and their conservatively substituted residues,

and Xaa at position 7 is selected from D, R and their conservatively substituted residues

<400> 40

Ala Arg His Arg Ile Xaa Xaa Ser Pro Ser Arg Ala Phe Asp Ile

1 5 10 15

<210> 41

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 41 ADI-27278, ADI-30293, ADI-30296

LCDR1-Kabat, L24-34

<400> 41

Lys Ser Ser Gln Ser Val Leu Phe Ser Ser Asn Asn Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 42

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 42 ADI-27278, ADI-30293, ADI-30296

LCDR2-Kabat, L50-56

<400> 42

Trp Ala Ser Thr Arg Glu Ser

fifteen

<210> 43

<211> 8

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 43 ADI-27278, ADI-30293, ADI-30296

LCDR3 Kabat, L89-97

<400> 43

Gln Gln Ser Tyr Phe Phe Pro Thr

fifteen

<210> 44

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 44 ADI-27291 HCDR1 KABAT,

numbered positions H27-35B

<400> 44

Phe Thr Phe Ser Ser Tyr Ala Met Ser

fifteen

<210> 45

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 45 ADI-30283 HCDR1 KABAT,

numbered positions H27-35B

<400> 45

Phe Thr Phe Ser Pro Tyr Gly Met Ser

fifteen

<210> 46

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 46 ADI-30286, ADI-30288 HCDR1

KABAT, numbered positions H27-35B

<400> 46

Phe Thr Phe Gly Pro Tyr Gly Met Ser

fifteen

<210> 47

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 47, consensus sequence

<220>

<221> misc_feature

<222> (4)..(5)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 4 is selected from S, G and their conservatively substituted residues,

and Xaa at position 5 is selected from S, P and their conservatively substituted residues

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from A, G, and its conservatively substituted residue

<400> 47

Phe Thr Phe Xaa Xaa Tyr Xaa Met Ser

fifteen

<210> 48

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 48 ADI-27291 HCDR2 KABAT, H50-65

<400> 48

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

1 5 10 15

gly

<210> 49

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 49 ADI-30283 HCDR2 KABAT, H50-65

<400> 49

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

1 5 10 15

gly

<210> 50

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 50 ADI-30286 HCDR2 KABAT, H50-65

<400> 50

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

1 5 10 15

gly

<210> 51

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 51 ADI-30288 HCDR2 KABAT, H50-65

<400> 51

Ala Ile Ser Gly Ser Gly Ala Ser Thr Trp His Ala Asp Ser Val Lys

1 5 10 15

gly

<210> 52

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 52 sequence, consensus sequence

<220>

<221> misc_feature

<222> (1)..(1)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from A, S, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (7)..(8)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 7 is selected from G, A and their conservatively substituted residues,

and Xaa at position 8 is selected from S, R and their conservatively substituted residues

<220>

<221> misc_feature

<222> (10)..(11)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 10 is selected from Y, W and their conservatively substituted residues,

and Xaa at position 11 is selected from Y, H and their conservatively substituted residues

<400> 52

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

1 5 10 15

gly

<210> 53

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 53 ADI-27291, ADI-30283 HCDR3 Kabat, H93-102

<400> 53

Ala Lys Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg

1 5 10 15

Tyr

<210> 54

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 54 ADI-30286 HCDR3 Kabat, H93-102

<400> 54

Ala Lys Asp Pro Gly Thr His Tyr Ser Gly Tyr Tyr Tyr Ile Trp Arg

1 5 10 15

Tyr

<210> 55

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 55 ADI-30288 HCDR3 Kabat, H93-102

<400> 55

Ala Lys Asp Pro Gly Thr Asp Ser Thr Gly Tyr Tyr Tyr Ile Trp Arg

1 5 10 15

Tyr

<210> 56

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 56 sequence, consensus sequence

<220>

<221> misc_feature

<222> (7)..(9)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 7 is selected from D, H and their conservatively substituted residues,

Xaa at position 8 is selected from S, Y and their conservatively substituted residues,

and Xaa at position 9 is selected from S, T and their conservatively substituted residues

<400> 56

Ala Lys Asp Pro Gly Thr Xaa Xaa Xaa Gly Tyr Tyr Tyr Ile Trp Arg

1 5 10 15

Tyr

<210> 57

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 57 ADI-27291, ADI-30296, ADI-30286,

ADI-30288 LCDR1 Kabat, L24-34

<400> 57

Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala

1 5 10

<210> 58

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 58 ADI-27291, ADI-30283, ADI-30286,

ADI-30288 LCDR2 Kabat, L50-56

<400> 58

Ser Ala Ser Thr Arg Ala Thr

fifteen

<210> 59

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 59 ADI-27291, ADI-30283, ADI-30286,

ADI-30288 LCDR3 Kabat, L89-97

<400> 59

Gln Gln Tyr Val Pro His Pro Pro Phe Thr

1 5 10

<210> 60

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 60 ADI-27297 HCDR1 KABAT,

numbered positions H27-35B

<400> 60

Tyr Thr Phe Thr Ser Tyr Tyr Met His

fifteen

<210> 61

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 61 ADI-30272, ADI-30278 HCDR1

KABAT, numbered positions H27-35B

<400> 61

Tyr Thr Phe Thr Glu Tyr Tyr Met His

fifteen

<210> 62

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 62 sequence, consensus sequence

<220>

<221> misc_feature

<222> (5)..(5)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, E, and its conservatively substituted residue

<400> 62

Tyr Thr Phe Thr Xaa Tyr Tyr Met His

fifteen

<210> 63

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 63 ADI-30272 HCDR2 KABAT, H50-65

<400> 63

Ile Ile Ser Pro Ser Ala Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln

1 5 10 15

gly

<210> 64

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 64 ADI-30278 HCDR2 KABAT, H50-65

<400> 64

Ile Ile Ser Pro Ser Ala Gly Ser Thr Lys Tyr Ala Gln Lys Phe Gln

1 5 10 15

gly

<210> 65

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 65 sequence, consensus sequence

<220>

<221> misc_feature

<222> (3)..(3)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from N, S, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(6)..(6)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from G, A, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (10)..(10)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, K, and its conservatively substituted residue

<400> 65

Ile Ile Xaa Pro Ser Xaa Gly Ser Thr Xaa Tyr Ala Gln Lys Phe Gln

1 5 10 15

gly

<210> 66

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 66 ADI-27297, ADI-30272 HCDR3 Kabat, H93-102

<400> 66

Ala Arg Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr

1 5 10 15

<210> 67

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 67 ADI-30278 HCDR3 Kabat, H93-102

<400> 67

Ala Arg Asp His Asp Ile Arg Leu Ala Gly Arg Leu Ala Asp Tyr

1 5 10 15

<210> 68

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 68 sequence, consensus sequence

<220>

<221> misc_feature

<222> (7)..(8)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 7 is selected from A, R and their conservatively substituted residues,

and Xaa at position 8 is selected from A, L and their conservatively substituted residues

<400> 68

Ala Arg Asp His Asp Ile Xaa Xaa Ala Gly Arg Leu Ala Asp Tyr

1 5 10 15

<210> 69

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 69 ADI-27297, ADI-30272, ADI-30278

LCDR1-Kabat, L24-34

<400> 69

Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala

1 5 10

<210> 70

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 70 ADI-27297, ADI-30272, ADI-30278

LCDR3 Kabat, L89-97

<400> 70

Gln Gln Ala Val Ile Leu Pro Ile Thr

fifteen

<210> 71

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 71 ADI-27301 HCDR1 KABAT,

numbered positions H27-35B

<400> 71

Gly Ser Ile Ser Ser Ser Ser Tyr Tyr Trp Gly

1 5 10

<210> 72

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 72 ADI-30306 HCDR1 KABAT,

numbered positions H27-35B

<400> 72

Gly Ser Ile Ser Ser Ser Leu Tyr Tyr Trp Gly

1 5 10

<210> 73

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 73 ADI-30311 HCDR1 KABAT,

numbered positions H27-35B

<400> 73

Gly Ser Ile Ala Ser Ser Val Tyr Tyr Trp Gly

1 5 10

<210> 74

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 74 consensus sequence

<220>

<221> misc_feature

<222> (4)..(4)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from A, S, and its conservatively substituted residue

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from S, L, V, and its conservatively substituted residue

<400> 74

Gly Ser Ile Xaa Ser Ser Xaa Tyr Tyr Trp Gly

1 5 10

<210> 75

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 75 ADI-30306 HCDR2 KABAT, H50-65

<400> 75

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

1 5 10 15

<210> 76

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 76 ADI-30311 HCDR2 KABAT, H50-65

<400> 76

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

1 5 10 15

<210> 77

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 77 consensus sequence

<220>

<221> misc_feature

<222>(9)..(9)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from Y, F, W, and its conservatively substituted residue

<220>

<221> misc_feature

<222> (14)..(14)

<223> Xaa can be any naturally occurring amino acid, preferably

selected from L, F, and its conservatively substituted residue

<400> 77

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

1 5 10 15

<210> 78

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 78 ADI-27301, ADI-30306 HCDR3 Kabat, H93-102

<400> 78

Ala Arg Glu Ala Gly Arg Gly Thr Thr Gly Leu Leu Phe Asp Tyr

1 5 10 15

<210> 79

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 79 ADI-30311 HCDR3 Kabat, H93-102

<400> 79

Ala Arg Glu Ala Gly Arg Thr Gly Thr Gly Leu Leu Phe Asp Tyr

1 5 10 15

<210> 80

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 80 sequence, consensus sequence

<220>

<221> misc_feature

<222> (7)..(8)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa at position 7 is selected from G, T and their conservatively substituted residues,

and Xaa at position 8 is selected from T, G and their conservatively substituted residues

<400> 80

Ala Arg Glu Ala Gly Arg Xaa Xaa Thr Gly Leu Leu Phe Asp Tyr

1 5 10 15

<210> 81

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 81 ADI-27301, ADI-30306, ADI-30311

LCDR1 Kabat, L24-34

<400> 81

Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala

1 5 10

<210> 82

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 82 ADI-27301, ADI-30306, ADI-30311

LCDR2-Kabat, L50-56

<400> 82

Lys Ala Ser Ser Leu Glu Ser

fifteen

<210> 83

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 83 ADI-27301, ADI-30306, ADI-30311

LCDR3 Kabat, L89-97

<400> 83

Gln Gln Tyr Gly Ile Leu Pro Arg Thr

fifteen

<210> 84

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 84 ADI-27238 vh

<400> 84

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asn Pro Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Ala Arg Tyr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 85

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 85 ADI-30263 vh

<400> 85

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asp Pro Ser Gly Gly Arg Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 86

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 86 ADI-30267 vh

<400> 86

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 87

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 87 ADI-30268 vh

<400> 87

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Ala Arg Thr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 88

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 88 ADI-27243 vh

<400> 88

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 89

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 89 ADI-30302 vh

<400> 89

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Arg Ser Gly Gly Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 90

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 90 ADI-30336 vh

<400> 90

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 91

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 91 ADI-27278 vh

<400> 91

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 92

<211> 121

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 92 ADI-30293 vh

<400> 92

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Arg His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 93

<211> 121

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 93 ADI-30296 vh

<400> 93

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Arg His Arg Ile Gly Arg Ser Pro Ser Arg Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 94

<211> 124

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 94 ADI-27291 vh

<400> 94

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120

<210> 95

<211> 124

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 95 ADI-30283 vh

<400> 95

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120

<210> 96

<211> 124

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 96 ADI-30286 vh

<400> 96

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr His Tyr Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120

<210> 97

<211> 124

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 97 ADI-30288 vh

<400> 97

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

1 5 10 15

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

20 25 30

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

35 40 45

Ser Ala Ile Ser Gly Ser Gly Ala Ser Thr Trp His Ala Asp Ser Val

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Thr Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120

<210> 98

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 98 ADI-27297 vh

<400> 98

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asn Pro Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 99

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 99 ADI-30272 vh

<400> 99

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Ser Pro Ser Ala Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 100

<211> 122

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 100 ADI-30278 vh

<400> 100

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Ser Pro Ser Ala Gly Ser Thr Lys Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Arg Leu Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 101

<211> 123

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 101 ADI-27301 vh

<400> 101

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 102

<211> 123

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 102 ADI-30306 vh

<400> 102

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Phe Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 103

<211> 123

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 103 ADI-30311 vh

<400> 103

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Trp Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 104

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 104 ADI-27238, ADI-30263, ADI-30267,

ADI-30268 VL

<400> 104

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 105

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 105 ADI-27243, ADI-30302 VL

<400> 105

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Ala Phe Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 106

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 106 ADI-30336 VL

<400> 106

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Ala His Pro Tyr

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 107

<211> 112

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 107 ADI-27278, ADI-30293, ADI-30296 VL

<400> 107

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

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Phe Ser

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

<210> 108

<211> 108

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 108 ADI-27291, ADI-30283, ADI-30286,

ADI-30288 VL

<400> 108

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Val Pro His Pro Pro

85 90 95

Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 109

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 109 ADI-27297, ADI-30272, ADI-30278 VL

<400> 109

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Val Ile Leu Pro Ile

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 110

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 110 ADI-27301, ADI-30306, ADI-30311 VL

<400> 110

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Ile Leu Pro Arg

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 111

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 111 ADI-27238 HC

<400> 111

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asn Pro Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Ala Arg Tyr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 112

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 112 ADI-30263 HC

<400> 112

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asp Pro Ser Gly Gly Arg Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 113

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 113 ADI-30267 HC

<400> 113

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

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

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 114

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 114 ADI-30268 HC

<400> 114

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Ala Arg Thr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 115

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 115 ADI-27243 HC

<400> 115

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 116

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 116 ADI-30302 HC

<400> 116

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Arg Ser Gly Gly Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 117

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 117 ADI-30336 HC

<400> 117

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 118

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 118 ADI-27278 HC

<400> 118

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile Trp

100 105 110

Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 119

<211> 447

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 119 ADI-30293 HC

<400> 119

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Arg His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

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

435 440 445

<210> 120

<211> 447

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 120 ADI-30296 HC

<400> 120

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Arg His Arg Ile Gly Arg Ser Pro Ser Arg Ala Phe Asp Ile Trp Gly

100 105 110

Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

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

435 440 445

<210> 121

<211> 450

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 121 ADI-27291 HC

<400> 121

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

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

180 185 190

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

195 200 205

Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser

210 215 220

Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly

225 230 235 240

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

245 250 255

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

260 265 270

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Leu Gly

450

<210> 122

<211> 450

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 122 ADI-30283 HC

<400> 122

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

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

180 185 190

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

195 200 205

Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser

210 215 220

Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly

225 230 235 240

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

245 250 255

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

260 265 270

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Leu Gly

450

<210> 123

<211> 450

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 123 ADI-30286 HC

<400> 123

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr His Tyr Ser Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

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

180 185 190

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

195 200 205

Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser

210 215 220

Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly

225 230 235 240

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

245 250 255

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

260 265 270

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Leu Gly

450

<210> 124

<211> 450

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 124 ADI-30288 HC

<400> 124

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

1 5 10 15

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

20 25 30

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

35 40 45

Ser Ala Ile Ser Gly Ser Gly Ala Ser Thr Trp His Ala Asp Ser Val

50 55 60

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

65 70 75 80

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

85 90 95

Ala Lys Asp Pro Gly Thr Asp Ser Thr Gly Tyr Tyr Tyr Ile Trp Arg

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

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

180 185 190

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

195 200 205

Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser

210 215 220

Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly

225 230 235 240

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

245 250 255

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

260 265 270

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Leu Gly

450

<210> 125

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 125 ADI-27297 HC

<400> 125

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Asn Pro Ser Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 126

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 126 ADI-30272 HC

<400> 126

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Ser Pro Ser Ala Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 127

<211> 448

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 127 ADI-30278 HC

<400> 127

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Ile Ile Ser Pro Ser Ala Gly Ser Thr Lys Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

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

85 90 95

Ala Arg Asp His Asp Ile Arg Leu Ala Gly Arg Leu Ala Asp Tyr Trp

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

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

435 440 445

<210> 128

<211> 449

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 128 ADI-27301 HC

<400> 128

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys

210 215 220

Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

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

435 440 445

gly

<210> 129

<211> 449

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 129 ADI-30306 HC

<400> 129

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Phe Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys

210 215 220

Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

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

435 440 445

gly

<210> 130

<211> 449

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 130 ADI-30311 HC

<400> 130

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

1 5 10 15

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

20 25 30

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

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Trp Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys

210 215 220

Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

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

435 440 445

gly

<210> 131

<211> 441

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 131 Antibody 10A7-IgG4 (Genentech) as a positive control HC

<400> 131

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

1 5 10 15

Ser Leu Lys Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser Ser Phe

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Arg Pro Leu Gly His Asn Thr Phe Asp Ser Trp Gly Gln Gly

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 132

<211> 457

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 132 Antibody 22G2-IgG4 (BMS) as a positive control HC

<400> 132

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Val Ser Ser Gly

20 25 30

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

35 40 45

Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser

50 55 60

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

65 70 75 80

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

85 90 95

Cys Ala Arg Asp Tyr Tyr Val Ser Gly Asn Tyr Tyr Asn Val Asp Tyr

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Lys Ser Leu Ser Leu Ser Leu Gly Lys

450 455

<210> 133

<211> 446

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 133 Antibody 31C6--IgG4 (MSD) as a positive control HC

<400> 133

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

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 Asp Pro Tyr Asn Asp Gly Ala 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 Gly Gly Pro Tyr Gly Trp Tyr Phe Asp Val Trp Gly Ala Gly

100 105 110

Thr Thr Val 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> 134

<211> 446

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 134 Antibody 1F4 in the role positive control HC

<400> 134

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

1 5 10 15

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

20 25 30

Leu Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Thr Ser Val 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> 135

<211> 467

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 135 IgG1 in the role negative control HC

<400> 135

Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala

65 70 75 80

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

85 90 95

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

100 105 110

Tyr Tyr Cys Ala Arg Gly Gly Pro Gly Trp Tyr Ala Ala Asp Val Trp

115 120 125

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

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

450 455 460

Ser Pro Gly

465

<210> 136

<211> 465

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 136 IgG4 in the role negative control HC

<400> 136

Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala

65 70 75 80

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

85 90 95

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

100 105 110

Tyr Tyr Cys Ala Arg Gly Gly Pro Gly Trp Tyr Ala Ala Asp Val Trp

115 120 125

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu

435 440 445

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

450 455 460

Lys

465

<210> 137

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 137 ADI-27238, ADI-30263, ADI-30267,

ADI-30268 LC

<400> 137

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 138

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 138 ADI-27243, ADI-30302 LC

<400> 138

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Ala Phe Pro Tyr

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 139

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 139 ADI-30336 LC

<400> 139

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Ala His Pro Tyr

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 140

<211> 219

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 140 ADI-27278, ADI-30293, ADI-30296 LC

<400> 140

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

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Phe Ser

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

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

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 141

<211> 215

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 141 ADI-27291, ADI-30283, ADI-30286,

ADI-30288 LC

<400> 141

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Val Pro His Pro Pro

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 142

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 142 ADI-27297, ADI-30272, ADI-30278 LC

<400> 142

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Val Ile Leu Pro Ile

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 143

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 143 ADI-27301, ADI-30306, ADI-30311 LC

<400> 143

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Ile Leu Pro Arg

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 144

<211> 220

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 144 Antibody 10A7-IgG4 as positive control LC

<400> 144

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Ile Thr Ser Val Gln Ala Glu Asp Met Gly Gln Tyr Phe Cys Gln Gln

85 90 95

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

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

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

180 185 190

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

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 145

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 145 Antibody 22G2-IgG4 (BMS) as a positive control LC

<400> 145

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

1 5 10 15

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

20 25 30

Leu Ala Trp Cys Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe Pro 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> 146

<211> 214

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 146 Antibody 31C6-IgG4 (MSD) as a positive control LC

<400> 146

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

1 5 10 15

Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu His Ile Tyr Ser Tyr

20 25 30

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

35 40 45

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

50 55 60

Ser Gly Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Pro

65 70 75 80

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

85 90 95

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

100 105 110

Pro Ser Val Phe Ile Phe 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> 147

<211> 219

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 147 Antibody 1F4 in the role positive control LC

<400> 147

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

1 5 10 15

Asp Gln Val Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val Asn Ser

20 25 30

Tyr Gly Asn Thr Phe Leu Ser Trp Tyr Leu His Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Phe Gly Ile Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

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

65 70 75 80

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

85 90 95

Thr His Gln Pro Pro Thr Phe Gly Pro Gly Thr Lys Leu Glu Val Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 148

<211> 236

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 148 IgG1 in the role negative control LC

<400> 148

Met Asp Phe Gln Val Gln Ile Ile Ser Phe Leu Leu Ile Ser Ala Ser

1 5 10 15

Val Ile Met Ser Arg Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

20 25 30

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

35 40 45

Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys

50 55 60

Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val

65 70 75 80

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

85 90 95

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

100 105 110

Ala Asp Leu Pro Ala Phe Ala Phe Gly Gly Gly Thr Lys Val Glu Ile

115 120 125

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

130 135 140

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

145 150 155 160

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

165 170 175

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

180 185 190

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

195 200 205

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

210 215 220

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 149

<211> 236

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 149 IgG4 in the role negative control LC

<400> 149

Met Asp Phe Gln Val Gln Ile Ile Ser Phe Leu Leu Ile Ser Ala Ser

1 5 10 15

Val Ile Met Ser Arg Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

20 25 30

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

35 40 45

Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys

50 55 60

Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val

65 70 75 80

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

85 90 95

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

100 105 110

Ala Asp Leu Pro Ala Phe Ala Phe Gly Gly Gly Thr Lys Val Glu Ile

115 120 125

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

130 135 140

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

145 150 155 160

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

165 170 175

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

180 185 190

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

195 200 205

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

210 215 220

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 150

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 150 ADI-27238 DNA VH

<400> 150

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcacc agctactata tgtcatgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcaacccta gtggtggtag cacaagctac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagggctaga 300

tacccatcaa gctggccata cggaatggac gtatggggcc agggaacaac tgtcaccgtc 360

tcctca 366

<210> 151

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 151 ADI-30263 DNA VH

<400> 151

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata cacattcctg agctactata tgaattgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcgatccta gtggtggtag gacaagctac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagggctaga 300

tacccatcaa gctggccata cggaacggat gtatggggcc agggaacaac tgtcaccgtc 360

tcctca 366

<210> 152

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 152 ADI-30267 DNA VH

<400> 152

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttccgg agctactata tgagttgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcgatccta gtggtggtcg tacaagcttc 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagggctaga 300

tacccagagt cttggccata cggaatggac gtatggggcc agggaacaac tgtcaccgtc 360

tcctca 366

<210> 153

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 153 ADI-30268 DNA VH

<400> 153

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcggg agctactata tggggtgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcgacccta gtggtggtag gacaagttac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagggctcgt 300

acgccatcaa gctggccata cggaatggac gtatggggcc agggaacaac tgtcaccgtc 360

tcctca 366

<210> 154

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 154 ADI-27243 DNA VH

<400> 154

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcagc agtagtcgct actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggagtatct attatagtgg gagcacctac 180

tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagat 300

gggttgtacc acaccccaga atacttccaa cactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 155

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 155 ADI-30302 DNA VH

<400> 155

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcggg agtagtcagt actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggagtatct atcgtagtgg ggggacctac 180

tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagat 300

gggttgtacc acaccccaga atacttccaa cactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 156

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 156 ADI-30336 DNA VH

<400> 156

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcagc agtagtcgct actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggagtatct attatagtgg gagcacctac 180

tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagat 300

gggttgtacc acaccccaga atacttccaa cactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 157

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 157 ADI-27278 DNA VH

<400> 157

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtggaa gtagtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagagaca atgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggcggtgt actactgcgc cagacaccgc 300

atagcagact caccaagcag agccttcgat atatggggtc agggtacaat ggtcaccgtc 360

tcctca 366

<210> 158

<211> 363

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 158 ADI-30293 DNA VH

<400> 158

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120

ccagggaagg gactggagtg ggtttcatac attagtagta gtgggaccat aaattacgca 180

gactctgtga agggccgatt caccatctcc agagacaatg ccaagaactc actgtatctg 240

caaatgaaca gcctgagagc cgaggacacg gcggtgtact actgcgccag acaccgcata 300

gcagactcac caagcagagc cttcgatata tggggtcagg gtacaatggt caccgtctcc 360

tca 363

<210> 159

<211> 363

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 159 ADI-30296 DNA VH

<400> 159

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccatcggg ggttatagca tgaactgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtagtaccat acattacgca 180

gactctgtga agggccgatt caccatctcc agagacaatg ccaagaactc actgtatctg 240

caaatgaaca gcctgagagc tgaggacacg gcggtgtact actgcgccag acaccgcata 300

gggcgttcac caagcagagc cttcgatata tggggtcagg gtacaatggt caccgtctcc 360

tca 363

<210> 160

<211> 372

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 160 ADI-27291 DNA VH

<400> 160

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggcggtgt actactgcgc caaggacccc 300

ggaacagaca gcagcggcta ctactacata tggcgatact ggggacaggg tacattggtc 360

accgtctcct ca 372

<210> 161

<211> 372

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 161 ADI-30283 DNA VH

<400> 161

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc ccgtatggga tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcaagt attagtggaa gtggtggtag gacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggcggtgt actactgcgc caaggacccc 300

ggaacagaca gcagcggcta ctactacata tggcgatact ggggacaggg tacattggtc 360

accgtctcct ca 372

<210> 162

<211> 372

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 162 ADI-30286 DNA VH

<400> 162

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttggc ccgtatggta tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggaa gtggtgcgag cacatggtac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca actccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggcggtgt actactgcgc caaggacccc 300

ggaacacatt atagcggcta ctactacata tggcgatact ggggacaggg tacattggtc 360

accgtctcct ca 372

<210> 163

<211> 372

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 163 ADI-30288 DNA VH

<400> 163

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttggc ccgtatggta tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggaa gtggtgcgag cacatggcac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggcggtgt actactgcgc caaggacccc 300

ggaacagaca gcactgggta ctactacata tggcgatact ggggacaggg tacattggtc 360

accgtctcct ca 372

<210> 164

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 164 ADI-27297 DNA VH

<400> 164

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcacc agctactata tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atcaacccta gtggtggtag cacaagctac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagagaccac 300

gacatagcag cagcaggaag attggctgat tactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 165

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 165 ADI-30272 DNA VH

<400> 165

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcacg gagtactata tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atctcgccta gtgcgggtag cacaagctac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagagaccac 300

gacatagcag cagcaggaag attggctgat tactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 166

<211> 366

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 166 ADI-30278 DNA VH

<400> 166

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg catctggata caccttcacg gagtactata tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaata atctcgccta gtgcgggtag cacaaagtac 180

240

atggagctga gcagcctgag atctgaggac acggcggtgt actactgcgc aagagaccac 300

gacatacgtc tggcaggaag attggctgat tactggggac agggtacatt ggtcaccgtc 360

tcctca 366

<210> 167

<211> 369

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 167 ADI-27301 DNA VH

<400> 167

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcagc agtagtagtt actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggagtatct attatagtgg gagcacctac 180

tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagaa 300

gccggacgcg gcaccactgg tctcttgttc gactactggg gacagggtac attggtcacc 360

gtctcctca 369

<210> 168

<211> 369

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 168 ADI-30306 DNA VH

<400> 168

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcagc agtagtcttt actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggtctatct attatagtgg gagcaccttt 180

tacaacccgt ccttcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagaa 300

gccggacgcg gcaccactgg tctcttgttc gactactggg gacagggtac attggtcacc 360

gtctcctca 369

<210> 169

<211> 369

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 169 ADI-30311 DNA VH

<400> 169

cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcgcg agtagtgttt actactgggg ctggatccgc 120

cagcccccag ggaaggggct ggagtggatt gggagtatct attatagtgg gagcacctgg 180

tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa gaaccagttc 240

tccctgaagc tgagttctgt gaccgccgca gacacggcgg tgtactactg cgccagagaa 300

gccggacgca ctgggactgg tctcttgttc gactactggg gacaggggtac attggtcacc 360

gtctcctca 369

<210> 170

<211> 321

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 170 ADI-27238, ADI-30263, ADI-30267,

ADI-30268 DNA VL

<400> 170

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gagcattaac agctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcagcaa agcctcctca ctcctttcac ttttggcgga 300

gggaccaagg ttgagatcaa a 321

<210> 171

<211> 321

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 171 ADI-27243, ADI-30302 DNA VL

<400> 171

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag agattcgcct tcccttacac ttttggcgga 300

gggaccaagg ttgagatcaa a 321

<210> 172

<211> 321

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 172 ADI-30336 DNA VL

<400> 172

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtttatta ctgtcagcag agattcgccc atccttacac ttttggcgga 300

gggaccaagg ttgagatcaa a 321

<210> 173

<211> 336

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 173 ADI-27278, ADI-30293,

ADI-30296 DNA VL

<400> 173

gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 60

atcaactgca agtccagcca gagtgtttta ttcagctcca acaataagaa ctacttagct 120

tggtaccagc agaaaccagg acagcctcct aagctgctca tttactgggc atctacccgg 180

gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 240

atcagcagcc tgcaggctga agatgtggca gtttattact gtcagcagtc ctacttcttc 300

cctacttttg gcggagggac caaggttgag atcaaa 336

<210> 174

<211> 324

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 174 ADI-27291, ADI-30283, ADI-30286,

ADI-30288 DNA VL

<400> 174

gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120

ggccaggctc ccaggctcct catctatagc gcatccacca gggccactgg tatcccagcc 180

aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240

gaagattttg cagtttatta ctgtcagcag tacgtccccc accctccttt cacttttggc 300

ggagggacca aggttgagat caaa 324

<210> 175

<211> 321

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 175 ADI-27297, ADI-30272, ADI-30278 DNA VL

<400> 175

gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 60

atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctccgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtcagcag gcagtcatcc tccctatcac ttttggcgga 300

gggaccaagg ttgagatcaa a 321

<210> 176

<211> 321

<212> DNA

<213> artificial sequence

<220>

<223> Antibody sequence 176 ADI-27301, ADI-30306, ADI-30311 DNA VL

<400> 176

gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctataaa gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttatta ctgccagcag tacggcatcc tccctaggac ttttggcgga 300

gggaccaagg ttgagatcaa a 321

<210> 177

<211> 327

<212> PRT

<213> artificial sequence

<220>

<223> Antibody 177 Heavy chain constant region (IgG4_PAA)

<220>

<221> misc_feature

<222> (327)..(327)

<223> Xaa can be any naturally occurring amino acid, and preferably

Xaa is lysine or deleted

<400> 177

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro 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 Xaa

325

<210> 178

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 178 ADI-27238 HCDR1-AbM

<400> 178

Gly Tyr Thr Phe Thr Ser Tyr Tyr Met Ser

1 5 10

<210> 179

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 179 ADI-30263 HCDR1-AbM

<400> 179

Gly Tyr Thr Phe Leu Ser Tyr Tyr Met Asn

1 5 10

<210> 180

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 180 ADI-30267 HCDR1-AbM

<400> 180

Gly Tyr Thr Phe Arg Ser Tyr Tyr Met Ser

1 5 10

<210> 181

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 181 ADI-30268 HCDR1-AbM

<400> 181

Gly Tyr Thr Phe Gly Ser Tyr Tyr Met Gly

1 5 10

<210> 182

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 182 ADI-27238 HCDR3-Kabat

<400> 182

Ala Arg Tyr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val

1 5 10

<210> 183

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 183 ADI-30263 HCDR3-Kabat

<400> 183

Ala Arg Tyr Pro Ser Ser Trp Pro Tyr Gly Thr Asp Val

1 5 10

<210> 184

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 184 ADI-30267 HCDR3-Kabat

<400> 184

Ala Arg Tyr Pro Glu Ser Trp Pro Tyr Gly Met Asp Val

1 5 10

<210> 185

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 185 ADI-30268 HCDR3-Kabat

<400> 185

Ala Arg Thr Pro Ser Ser Trp Pro Tyr Gly Met Asp Val

1 5 10

<210> 186

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 186 ADI-27243, ADI-30336 HCDR1-AbM

<400> 186

Gly Gly Ser Ile Ser Ser Ser Arg Tyr Tyr Trp Gly

1 5 10

<210> 187

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 187 ADI-30302 HCDR1-AbM

<400> 187

Gly Gly Ser Ile Gly Ser Ser Gln Tyr Tyr Trp Gly

1 5 10

<210> 188

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 188 ADI-27243, ADI-30302, ADI-30336

HCDR3-Kabat

<400> 188

Asp Gly Leu Tyr His Thr Pro Glu Tyr Phe Gln His

1 5 10

<210> 189

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 189 ADI-27278, ADI-30293 HCDR1-AbM

<400> 189

Gly Phe Thr Phe Ser Ser Tyr Ser Met Asn

1 5 10

<210> 190

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 190 ADI-30296 HCDR1-AbM

<400> 190

Gly Phe Thr Ile Gly Gly Tyr Ser Met Asn

1 5 10

<210> 191

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 191 ADI-27278, ADI-30293 HCDR3-Kabat

<400> 191

His Arg Ile Ala Asp Ser Pro Ser Arg Ala Phe Asp Ile

1 5 10

<210> 192

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 192 ADI-30296 HCDR3-Kabat

<400> 192

His Arg Ile Gly Arg Ser Pro Ser Arg Ala Phe Asp Ile

1 5 10

<210> 193

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 193 ADI-27291 HCDR1-AbM

<400> 193

Gly Phe Thr Phe Ser Ser Tyr Ala Met Ser

1 5 10

<210> 194

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 194 ADI-30283 HCDR1-AbM

<400> 194

Gly Phe Thr Phe Ser Pro Tyr Gly Met Ser

1 5 10

<210> 195

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 195 ADI-30286, ADI-30288 HCDR1-AbM

<400> 195

Gly Phe Thr Phe Gly Pro Tyr Gly Met Ser

1 5 10

<210> 196

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 196 ADI-27291, ADI-30283 HCDR3-Kabat

<400> 196

Asp Pro Gly Thr Asp Ser Ser Gly Tyr Tyr Tyr Ile Trp Arg Tyr

1 5 10 15

<210> 197

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 197 ADI-30286 HCDR3-Kabat

<400> 197

Asp Pro Gly Thr His Tyr Ser Gly Tyr Tyr Tyr Ile Trp Arg Tyr

1 5 10 15

<210> 198

<211> 15

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 198 ADI-30288 HCDR3-Kabat

<400> 198

Asp Pro Gly Thr Asp Ser Thr Gly Tyr Tyr Tyr Ile Trp Arg Tyr

1 5 10 15

<210> 199

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 199 ADI-27297 HCDR1-AbM

<400> 199

Gly Tyr Thr Phe Thr Ser Tyr Tyr Met His

1 5 10

<210> 200

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 200 ADI-30272, ADI-30278 HCDR1-AbM

<400> 200

Gly Tyr Thr Phe Thr Glu Tyr Tyr Met His

1 5 10

<210> 201

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 201 ADI-27297, ADI-30272 HCDR3-Kabat

<400> 201

Asp His Asp Ile Ala Ala Ala Gly Arg Leu Ala Asp Tyr

1 5 10

<210> 202

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 202 ADI-30278 HCDR3-Kabat

<400> 202

Asp His Asp Ile Arg Leu Ala Gly Arg Leu Ala Asp Tyr

1 5 10

<210> 203

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 203 ADI-27301 HCDR1-AbM

<400> 203

Gly Gly Ser Ile Ser Ser Ser Ser Tyr Tyr Trp Gly

1 5 10

<210> 204

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 204 ADI-30306 HCDR1-AbM

<400> 204

Gly Gly Ser Ile Ser Ser Ser Leu Tyr Tyr Trp Gly

1 5 10

<210> 205

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> Sequence of antibody 205 ADI-30311 HCDR1-AbM

<400> 205

Gly Gly Ser Ile Ala Ser Ser Val Tyr Tyr Trp Gly

1 5 10

<210> 206

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 206 ADI-27301, ADI-30306 HCDR3-Kabat

<400> 206

Glu Ala Gly Arg Gly Thr Thr Gly Leu Leu Phe Asp Tyr

1 5 10

<210> 207

<211> 13

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 207 ADI-30311 HCDR3-Kabat

<400> 207

Glu Ala Gly Arg Thr Gly Thr Gly Leu Leu Phe Asp Tyr

1 5 10

<210> 208

<211> 244

<212> PRT

<213> artificial sequence

<220>

<223> Sequence 208 human TIGIT sequence

<400> 208

Met Arg Trp Cys Leu Leu Leu Ile Trp Ala Gln Gly Leu Arg Gln Ala

1 5 10 15

Pro Leu Ala Ser Gly Met Met Thr Gly Thr Ile Glu Thr Thr Gly Asn

20 25 30

Ile Ser Ala Glu Lys Gly Gly Ser Ile Ile Leu Gln Cys His Leu Ser

35 40 45

Ser Thr Thr Ala Gln Val Thr Gln Val Asn Trp Glu Gln Gln Asp Gln

50 55 60

Leu Leu Ala Ile Cys Asn Ala Asp Leu Gly Trp His Ile Ser Pro Ser

65 70 75 80

Phe Lys Asp Arg Val Ala Pro Gly Pro Gly Leu Gly Leu Thr Leu Gln

85 90 95

Ser Leu Thr Val Asn Asp Thr Gly Glu Tyr Phe Cys Ile Tyr His Thr

100 105 110

Tyr Pro Asp Gly Thr Tyr Thr Gly Arg Ile Phe Leu Glu Val Leu Glu

115 120 125

Ser Ser Val Ala Glu His Gly Ala Arg Phe Gln Ile Pro Leu Leu Gly

130 135 140

Ala Met Ala Ala Thr Leu Val Val Ile Cys Thr Ala Val Ile Val Val

145 150 155 160

Val Ala Leu Thr Arg Lys Lys Lys Ala Leu Arg Ile His Ser Val Glu

165 170 175

Gly Asp Leu Arg Arg Lys Ser Ala Gly Gln Glu Glu Glu Trp Ser Pro Ser

180 185 190

Ala Pro Ser Pro Pro Gly Ser Cys Val Gln Ala Glu Ala Ala Pro Ala

195 200 205

Gly Leu Cys Gly Glu Gln Arg Gly Glu Asp Cys Ala Glu Leu His Asp

210 215 220

Tyr Phe Asn Val Leu Ser Tyr Arg Ser Leu Gly Asn Cys Ser Phe Phe

225 230 235 240

Thr Glu Thr Gly

<210> 209

<211> 107

<212> PRT

<213> artificial sequence

<220>

<223> Antibody sequence 209 The constant region of the lung chains (? light chain)

<400> 209

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

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Claims (55)

1. An antibody or antigen-binding fragment thereof that binds to TIGIT and contains three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), where: (a) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 1, 2, 3, 4 or 5, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 6, 7, 8, 9 or 10, HCDR3 contains the amino acid sequence, presented in SEQ ID NO: 11, 12, 13, 14 or 15, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 16, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 17, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 18; (b) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 19, 20 or 21, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 22, 23 or 24, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 25, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 26, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 27, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 28, 29 or 30; (c) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 31, 32 or 33, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 34, 35, 36 or 37, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 38, 39 or 40, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 41, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 42, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 43; (d) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 44, 45, 46 or 47, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 48, 49, 50, 51 or 52, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 48, 49, 50, 51 or 52 SEQ ID NO: 53, 54, 55 or 56, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 57, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 58, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 59; or (e) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 60, 61 or 62, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 6, 63, 64 or 65, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 66, 67 or 68, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 69, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 17, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 70; (f) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 71, 72, 73 or 74, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 22, 75, 76 or 77, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 78, 79 or 80, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 81, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 82, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 83; (g) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 178, 179, 180 or 181, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 6, 7, 8 or 9, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 182, 183, 184 or 185, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 16, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 17, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 18; (h) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 186 or 187, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 22 or 23, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 188, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 188, LCDR1 contains the amino acid sequence presented in SEQ ID NO: 26, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 27, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 28 or 29; (i) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 189 or 190, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 34, 35 or 36, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 191 or 192, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 41, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 42, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 43; (j) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 193, 194 or 195, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 48, 49, 50 or 51, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 196, 197 or 198, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 57, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 58, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 59; (k) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 199 or 200, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 6, 63 or 64, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 201 or 202, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 69, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 17, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 70; or (l) HCDR1 contains the amino acid sequence shown in SEQ ID NO: 203, 204 or 205, HCDR2 contains the amino acid sequence shown in SEQ ID NO: 22, 75 or 76, HCDR3 contains the amino acid sequence shown in SEQ ID NO: 206 or 207, LCDR1 contains the amino acid sequence shown in SEQ ID NO: 81, LCDR2 contains the amino acid sequence shown in SEQ ID NO: 82, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 83. 2. An antibody or antigen-binding fragment according to claim 1, containing: (i) a heavy chain variable region comprising an amino acid sequence that is at least 80%, 85%, or 90% identical to the amino acid sequence shown in SEQ ID NO: 84, 85, 86, or 87, and/or a light chain variable region a chain containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 104; (ii) a heavy chain variable region containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 88, 89 or 90, and/or a light chain variable region containing an amino acid sequence that is at least 80%, 85%, or 90% identical to the amino acid sequence shown in SEQ ID NO: 105 or 106; (iii) a heavy chain variable region containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 91, 92 or 93, and/or a light chain variable region containing an amino acid sequence that is at least 80%, 85%, or 90% identical to the amino acid sequence shown in SEQ ID NO: 107; (iv) a heavy chain variable region comprising an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 94, 95, 96 or 97 and/or a light chain variable region containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 108; (v) a heavy chain variable region containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 98, 99 or 100, and/or a light chain variable region containing an amino acid sequence that is at least 80%, 85%, or 90% identical to the amino acid sequence shown in SEQ ID NO: 109; or (vi) a heavy chain variable region containing an amino acid sequence that is at least 80%, 85% or 90% identical to the amino acid sequence shown in SEQ ID NO: 101, 102 or 103 and/or a light chain variable region containing an amino acid sequence that is at least 80%, 85%, or 90% identical to the amino acid sequence shown in SEQ ID NO: 110. 3. An antibody or antigen-binding fragment according to any one of paragraphs. 1-2 containing a heavy chain variable region and a light chain variable region selected from: (i) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 84, 85, 86 or 87 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 104; (ii) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 88, 89 or 90 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 105 or 106; (iii) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 91, 92 or 93 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 107; (iv) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 94, 95, 96, or 97 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 108; (v) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 98, 99 or 100 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 109; or (vi) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 101, 102 or 103 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 110. 4. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody is an IgG1, IgG2 or IgG4 isotype antibody, or an antigen-binding fragment thereof, preferably an IgG4 Fc region containing mutations S228P, F234A and L235A. 5. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody is a fully humanized antibody, a humanized antibody, or a chimeric antibody. 6. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antigen-binding fragment is an antibody fragment selected from: Fab, Fab', Fab'-SH, Fv, single chain antibodies such as scFv, (Fab') ₂ -fragments, single domain antibodies, diabodies (dAb) or linear antibodies. 7. An antibody or antigen-binding fragment thereof according to any one of the preceding claims, wherein the antibody has one or more of the following properties: (i) the ability to bind to human TIGIT with high affinity; (ii) the ability to cross-react immunologically with monkey and/or mouse TIGIT; (iii) efficient binding to TIGIT on the cell surface; (iv) blocking the binding of TIGIT to its CD155 ligand; (v) reducing the inhibitory effect of CD155 binding to TIGIT on IL-2 dependent downstream signaling; (vi) increased production of IL-2 by T-lymphocytes; (vii) antitumor activity, eg tumor growth inhibition; (viii) a more pronounced tumor inhibitory effect in combination with an anti-PD-1 antibody, eg, enhanced tumor growth inhibition. 8. An isolated nucleic acid encoding an anti-TIGIT antibody or antigen-binding fragment thereof according to any of the preceding claims. 9. An expression vector containing the nucleic acid according to claim 8. 10. A host cell for expressing an anti-TIGIT antibody or antigen-binding fragment thereof, containing the nucleic acid of claim 8 or the expression vector of claim 9. 11. A method for producing an anti-TIGIT antibody or antigen-binding fragment thereof, comprising culturing the host cell according to paragraph 10 under conditions suitable for the expression of a nucleic acid encoding an anti-TIGIT antibody or antigen-binding fragment according to any one of paragraphs. 1-7, and optionally isolating the antibody or antigen-binding fragment thereof, wherein optionally said method further comprises isolating the anti-TIGIT antibody or antigen-binding fragment thereof from the host cell. 12. Immunoconjugate to enhance the immune response in a subject containing an antibody or antigen-binding fragment according to any one of paragraphs. 1-7 conjugated with a therapeutic agent. 13. Immunoconjugate for the detection of TIGIT in the sample, containing the antibody or antigen-binding fragment according to any one of paragraphs. 1-7 conjugated with a diagnostic agent. 14. Multispecific antibody to enhance the immune response in a subject, comprising an antibody or antigen-binding fragment according to any one of paragraphs. 1-7, where the polyspecific antibody is preferably a bispecific antibody. 15. Pharmaceutical composition for enhancing the immune response in a subject, containing an antibody or antigen-binding fragment according to any one of paragraphs. 1-7, and a pharmaceutical excipient. 16. The pharmaceutical composition of claim 15, comprising a second therapeutic agent, wherein the second therapeutic agent is preferably selected from an anti-PD-1 antibody or an anti-PD-L1 antibody. 17. A method for preventing or treating a tumor in a subject, comprising administering to the subject an effective amount of an antibody to TIGIT or an antigen-binding fragment according to any one of paragraphs. 1-7 or a pharmaceutical composition according to claim 15 or 16. 18. The method of claim 17, wherein the tumor is a malignant neoplasm of the gastrointestinal tract, such as colon cancer. 19. A method for preventing or treating an infectious disease in a subject, comprising administering to the subject an effective amount of an antibody to TIGIT or an antigen-binding fragment according to any one of paragraphs. 1-7 or a pharmaceutical composition according to claim 15 or 16. 20. A method of blocking the binding of TIGIT to CD155 in order to reduce or eliminate the immunosuppressive effect of TIGIT in a subject, comprising administering to the subject an effective amount of an antibody to TIGIT or an antigen-binding fragment according to any one of paragraphs. 1-7 or a pharmaceutical composition according to claim 15 or 16. 21. A method for detecting TIGIT in a sample, including: (a) bringing the sample into contact with the antibody or antigen-binding fragment according to any one of paragraphs. 1–7; and (b) detection of the complex formed by the antibody or its antigen-binding fragment and TIGIT, where the antibody contains a detectable label.

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