Monoclonal antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor beta chain, and uses thereof

Abstract

This invention relates to the fields of biotechnology and medicine, particularly to monoclonal antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 region of the variable domain of the human T-cell receptor β chain. The invention further relates to nucleic acids encoding said antibodies, expression vectors, host cells, and methods for producing said antibodies; methods for producing antibodies according to the invention; pharmaceutical compositions comprising antibodies according to the invention; pharmaceutical compositions comprising antibodies according to the invention and other therapeutically active compounds; methods for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain; the use of antibodies or pharmaceutical compositions thereof for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain; and the use of antibodies and other therapeutically active compounds according to the invention for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain.

Description

Invention Field

[0001] This invention relates to the fields of biotechnology and medicine, particularly to monoclonal antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 region of the variable domain of the human T-cell receptor β chain. The invention further relates to nucleic acids encoding said antibodies, expression vectors, host cells, and methods for producing said antibodies; methods for producing antibodies according to the invention; pharmaceutical compositions comprising antibodies according to the invention; pharmaceutical compositions comprising antibodies according to the invention and other therapeutically active compounds; methods for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain; the use of antibodies or pharmaceutical compositions thereof for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain; and the use of antibodies and other therapeutically active compounds according to the invention for treating or preventing diseases or conditions mediated by the TRBV5-1 region of the variable domain of the human T-cell receptor β chain. Background of the Invention Monoclonal antibodies in chimeric, humanized, or fully human molecular forms have proven to be effective drugs for the treatment of a variety of conditions and diseases.

[0003] Autoimmune diseases are partly caused by autoreactive T lymphocytes (Haroon N et al., ArthritisRheum. Oct 2013; 65(10):2645-54., Duarte J et al., PloS One May 10, 2010; 5(5):e10558; Konig M et al., Front Immunol Jan 25, 2016; 7:11). The major role in the emergence of autoreactive T lymphocyte clones is exerted through the interaction of the antigen-recognizing T cell receptor (TCR) with proteins of the major histocompatibility complex (MHC, HLA), which presents processed peptides of intracellular proteins or those of pathogenic organisms on its surface.

[0004] Existing technologies provide that T cell receptor sequences are markers that allow for the identification of T lymphocyte clones involved in the pathogenesis of autoimmune diseases. T cell receptor subunits structurally belong to the immunoglobulin superfamily and are formed by several gene segments. The variable region of the TCR forms the antigen-binding center of the TCR. This means they are clonal specific, i.e., they differ in these T lymphocytes that respond to different antigens.

[0005] T lymphocytes (T cells) are stimulated when an antigen binds to its T cell receptor. The TCR, the defining structure of a T cell, is a transmembrane heterodimer composed of α and β chains or δ and γ chains linked by disulfide bonds. Within these chains are complementarity-determining regions (CDRs) that determine the antigens the TCR will bind to. TCR development occurs through a lymphocyte-specific process of genetic recombination, in which numerous potential segments assemble into the final sequence. This genetic recombination of TCR gene segments in somatic T cells occurs during the early stages of development within the thymus. The TCRα gene locus contains variable (V) and linked (J) gene segments (Vα and Jα), while the TCRβ locus contains D gene segments in addition to the Vβ and Jβ segments. Thus, the α chain is generated by VJ recombination, and the β chain involves VDJ recombination.

[0006] The TCR α-chain gene locus consists of 46 variable regions (TRAV), 8 linker regions (TRAJ), and constant regions. The TCR β-chain gene locus consists of 48 variable regions (TRBV), followed by two diversity regions (TRBD), 12 linker regions (TRBJ), and two constant regions. (Bio-Rad. Mini-review | An overview of T cell receptors [Electronic resource] / / Bio-Rad. URL: https: / / www.bio-rad-antibodies.com / t-cell-receptor-minireview.html).

[0007] The T-cell receptor interacts with an antigen that binds non-covalently to the MHC molecule via six complementarity-determining regions (CDRs): three α-chain regions and three β-chain regions. These CDRs represent variable regions, specifically the variable domains of the T-cell receptor, loop-Vα (TRAV) and Vβ (TRBV).

[0008] The relationship between TCR CD4+ T cells and the development of type 1 diabetes is known according to existing technology (Mai T. Tran ET ALL., T cell receptor recognition of hybrid insulin peptides bound to HLA-DQ8, NATURE COMMUNICATIONS, 2021, 12:5110, https: / / doi.org / 10.1038 / s41467-021-25404-x, Maki Nakayama ET ALL., Using the T Cell Receptor as a Biomarker in Type 1 Diabetes, Front. Immunol., 17 November 2021, Sec. Immunological Tolerance and Regulation, https: / / doi.org / 10.3389 / fimmu.2021.777788).

[0009] Patent document WO2020010250 provides an antibody that specifically binds to the variable region of the T cell receptor β chain (TCRβV).

[0010] Patent documents WO2019132738 and WO2020139171 provide antibodies that specifically bind to the β-chain variable domain region of human T-cell receptors belonging to the TRBV9 family.

[0011] Therefore, there is a real need to generate antibodies that specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0012] Summary of the Invention (Detailed Description of the Invention) The inventors of this invention have developed an antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain and exhibits a high binding affinity parameter for the TRBV5-1 segment of the human T cell receptor β chain. Furthermore, the antibody according to the invention is humanized, stable and thermostable in human serum, and also possesses antibody-dependent cell-mediated cytotoxicity (ADCC).

[0013] Definition and general method Unless otherwise expressly defined herein, all technical and scientific terms used in conjunction with this invention have the same meaning as commonly understood by those skilled in the art.

[0014] Furthermore, unless the context otherwise requires, singular terms should include plural terms, and plural terms should include singular terms. Generally, the cell culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, organic synthetic chemistry, medicinal chemistry, and protein and nucleic acid hybridization and chemistry methods described herein are well-known and widely used by those skilled in the art. Enzymatic reactions and purification methods are performed according to the manufacturer's guidelines, as are those commonly found in the art or as described herein.

[0015] The term “KD” in this specification refers to the affinity constant (or equilibrium constant), which is calculated from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M).

[0016] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise stated, "binding affinity" refers to the inherent (characteristic, true) binding affinity, which reflects a 1:1 interaction between the members of the binding pair (e.g., antibody and antigen). The affinity of molecule X for its binding partner Y can generally be expressed by an affinity constant (KD). Preferred Kd values ​​are about 200 nM, 150 nM, 100 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 8 nM, 6 nM, 4 nM, 2 nM, 1 nM, or lower. Affinity can be measured by common methods known in the art, including those described in this specification. Low-affinity antibodies generally bind antigens slowly and tend to dissociate easily, while high-affinity antibodies generally bind antigens more rapidly and tend to remain bound for longer. Various methods for measuring binding affinity are known in the art, and any one of these methods can be used for the purposes of this invention.

[0017] The terms “Kd,” “koff,” or “kdis” refer to the dissociation rate constant of a specific interaction between a binding molecule and an antigen. The dissociation rate constant koff can be measured using biomembrane interferometry techniques, such as the Octet™ system.

[0018] The terms “Ka,” “kon,” or “on-rate” refer to the association rate constant.

[0019] Term "R" 2 "Refers to the coefficient of determination."

[0020] The term "response" refers to the antibody-antigen binding signal.

[0021] The term "in vitro" refers to a biological entity, biological process, or biological reaction occurring outside an organism under artificial conditions. For example, cells grown in vitro should be understood as cells grown in an environment outside the organism, such as in test tubes, culture flasks, or microtiter plates.

[0022] The term "ED" 50 (EC) 50 (50% effective dose / concentration, half maximum effective concentration) refers to the concentration of a formulation that produces 50% of the biological effect (which may include cytotoxicity).

[0023] Human T-cell receptors are heterodimeric protein complexes located on the surface of T cells. These receptors are found only on T cells. The primary function of T-cell receptors is to specifically recognize processed antigens that bind to molecules of the major histocompatibility complex (MHC).

[0024] The terms “anti-TRBV5-1 antibody,” “antibody against TRBV5-1,” “antibody specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain,” and “antibody specifically binding to the TRBV5-1 segment” are used interchangeably in the context of this application and refer to an antibody specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0025] ADCC—Antibody-dependent cell-mediated cytotoxicity.

[0026] ADCP—Antibody-dependent phagocytosis.

[0027] NFAT—Activated T cell nuclear factor.

[0028] Luc—luciferase.

[0029] FBS—Fetal bovine serum.

[0030] CD3—Differentiation cluster 3.

[0031] CD16—Differentiation Cluster 16.

[0032] CD20—Differentiation Cluster 20.

[0033] CD64—Differentiation cluster 64.

[0034] PBMCs—peripheral blood mononuclear cells.

[0035] PE—phycoerythrin.

[0036] As used in this specification and the following claims, unless the context otherwise requires, the words “include” and “comprise”, or variations thereof such as “includes”, “including”, “comprises”, or “comprising”, shall be construed as implying inclusion of the said integer or group of integers, but not excluding any other integer or group of integers.

[0037] Antibody This invention relates to monoclonal antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0038] The term "monoclonal antibody" or "mAb" refers to an antibody synthesized and isolated from various clonal populations of cells.

[0039] The antibody of this invention is a recombinant antibody.

[0040] The term "recombinant antibody" refers to an antibody expressed in a cell or cell line containing a nucleotide sequence encoding an antibody that is not associated with that cell in nature.

[0041] In one aspect, the present invention relates to a monoclonal antibody or antigen-binding fragment thereof that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, comprising: (a) Contains the following light chain variable structural domains: (i) having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) CDR2 having the amino acid sequence EIX1KLX2S, wherein X1 = T or S; X2 = A, F, or M; and (iii) CDR3 having the amino acid sequence QQWNYPХ3LX4, in which X3 = L, R, or Y; X4 = S or T; as well as (b) Contains the following heavy-chain variable structural domains: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) CDR2 having the amino acid sequence YINPX5X6GRTGYNQKX7QX8, in which X5 = Y or W; X6 = Q or N; X7 = F or L; X8 = A or G; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0042] In one embodiment of the invention, the antibody according to the invention is an isolated antibody.

[0043] The term "isolated" used to describe various antibodies according to this specification refers to antibodies that have been identified, isolated, and / or regenerated from cells or cell cultures in which the antibody is expressed. Impurities (contaminating components) from the natural environment are materials that typically interfere with the diagnostic or therapeutic use of peptides and may include enzymes, hormones, and other protein or non-protein solutes. Isolated peptides are typically prepared through at least one purification step.

[0044] In one embodiment of the present invention, the antibody according to the present invention is a humanized antibody.

[0045] Humanized antibodies are defined as antibodies whose amino acid sequence is at least 95% homologous to the reference sequence of human immunoglobulins (according to the IMGT database). The use of humanized antibodies reduces the risk of adverse immune responses in the human body when they are used.

[0046] As used herein, the terms "antibody" or "immunoglobulin" (Ig) include complete antibodies. The term "antibody" refers to a glycoprotein comprising at least two heavy chains (H) and two light chains (L) linked together by disulfide bonds. Each heavy chain comprises a heavy chain variable region (hereinafter referred to as VH) and a heavy chain constant region. Each light chain consists of a light chain variable region (hereinafter referred to as VL) and a light chain constant region. The light chain constant domain may be CK (κ light chain constant domain) or CL (λ light chain constant domain). Preferably, the light chain is a kappa (κ) light chain, and the light chain constant domain is preferably CK.

[0047] The antibodies according to the invention can be any class (e.g., IgA, IgD, IgE, IgG and IgM, preferably IgG) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG1).

[0048] The VL and VH regions can be further subdivided into highly variable regions called complementarity-determining regions (CDRs), which are interspersed among more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen.

[0049] The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0050] As used in this specification, the term “antigen-binding portion” or “antigen-binding fragment” of an antibody refers to one or more antibody fragments that retain the ability to bind specifically to an antigen. The antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments included within the term “antigen-binding portion” of an antibody include: (i) Fab fragments, monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments connected by a disulfide bridge at the hinge region; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of VL and VH domains of a single arm of the antibody; and (v) dAb fragments consisting of VH / VHH domains (Ward et al., (1989) Nature 341:544-546). Additionally, the two regions of the Fv fragment, VL and VH, are encoded by different genes and can be linked using a synthetic adapter via recombination methods. This adapter allows them to act as a single protein linker, where the VL and VH regions pair to form a monovalent molecule (called a single-chain Fv (scFv); see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). It is assumed that such single-chain molecules are also included within the term "antigen-binding portion" of an antibody. Such antibody fragments are produced using conventional techniques known to those skilled in the art, and these fragments are screened in the same manner as intact antibodies.

[0051] As used in this application, "Kabat numbering scheme" or "numbering according to Kabat" refers to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the variable regions of the heavy and light chains of the antibody (Kabat et al. Ann. NY Acad. Sci., 190:382-93 (1971); Kabat et al. Sequences of Proteins of Immunological Interest, 5th edition, US Department of Health and Human Services, NIH Publication No. 91-3242 (1991)).

[0052] The antibodies of the present invention that "specifically bind" to a target antigen refer to antibodies that bind to the antigen with sufficient affinity, such that the antibody can be used as a diagnostic and / or therapeutic agent targeting proteins, cells, or tissues expressing the antigen, and that have slight cross-reactivity with other proteins.

[0053] The term “specific binding” to an epitope on a specific polypeptide or a specific target polypeptide can be described for example for a molecule that has the following Kd: at least about 200 nM, or at least about 150 nM, or at least about 100 nM, or at least about 60 nM, or at least about 50 nM, or at least about 40 nM, or at least about 30 nM, or at least about 20 nM, or at least about 10 nM, or at least about 8 nM, or at least about 6 nM, or at least about 4 nM, or at least about 2 nM, or at least about 1 nM or less.

[0054] In one implementation, the term "specific binding" refers to a binding in which a molecule binds to a specific polypeptide or an epitope on a specific polypeptide, without fundamental binding to any other polypeptide or an epitope on a polypeptide.

[0055] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a light chain variable domain comprising a CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7.

[0056] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a light chain variable domain comprising a CDR3 having an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11.

[0057] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a light chain variable domain comprising the following: (i) having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7; as well as (iii) Having a CDR3 with an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11.

[0058] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (i) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (ii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (iii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (iv) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (v) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (vi) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (vii) contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 10; or (viii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; or (ix) contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 10; or (x) contains the following light chain variable structure domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 9.

[0059] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a heavy chain variable domain comprising CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17.

[0060] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a heavy chain variable domain comprising the following: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0061] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (i) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ii) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iii) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iv) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (v) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0062] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (a) Contains the following light chain variable structural domains: (i) having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7; as well as (iii) A CDR3 having an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0063] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (i) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (v) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (vi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (vii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (viii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ix) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (x) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xiii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 10; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xiv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xvi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xvii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xviii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ixx) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 10; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xx) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0064] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a light chain variable domain comprising an amino acid sequence selected from the following: SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27 or SEQ ID NO: 28.

[0065] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment includes a heavy chain variable domain comprising an amino acid sequence selected from the following: SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO: 34.

[0066] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (a) A light chain variable domain comprising an amino acid sequence selected from the following: SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27 or SEQ ID NO: 28; and (b) A heavy chain variable domain comprising an amino acid sequence selected from the following: SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO: 34.

[0067] In some embodiments of the present invention, the monoclonal antibody or its antigen-binding fragment comprises: (i) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 29; or (ii) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 29; or (iii) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 21, and (b) A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30; or (iv) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22, and (b) A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30; or (v) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 23, and (b) A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 30; or (vi) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 23, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 31; or (vii) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32; or (viii) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32; or (ix) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32; or (x) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 26, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32; or (xi) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 21, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xii) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xiii) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 25, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34; or (xiv) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 20, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xv) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 22, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xvi) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xvii) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (xviii) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 23, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33; or (ixx) (a) A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 27, and (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34; or (xx) (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 28, and (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34.

[0068] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is a full-length IgG antibody.

[0069] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is a full-length IgG antibody having human IgG1, IgG2, IgG3 or IgG4 isotypes.

[0070] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is a full-length IgG antibody having the human IgG1 isotype.

[0071] In some embodiments of the invention, the monoclonal antibody contains, in the Fc fragment, mutations S239D and I332E in the CH2 region according to the EU numbering scheme of the antibody amino acids (Edelman GM et al., Proc. Natl. Acad. Sci. USA 63 (1969) pp. 78-85; Kabat, EA, et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, (1991).

[0072] In some embodiments of the invention, the monoclonal antibody contains G236A, S239D, and I332E mutations in the CH2 region according to the EU numbering scheme of the antibody amino acids.

[0073] In some embodiments of the invention, the monoclonal antibody contains deletions of 446G and 447K in the CH3 region according to the EU numbering scheme of the antibody amino acids.

[0074] In some embodiments of the present invention, the monoclonal antibody comprises a light chain containing an amino acid sequence selected from the following: SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 or SEQ ID NO: 44.

[0075] In some embodiments of the present invention, the monoclonal antibody comprises a heavy chain containing an amino acid sequence selected from the following: SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 or SEQ ID NO: 52.

[0076] In some embodiments of the present invention, the monoclonal antibody includes: (i) (a) A light chain comprising an amino acid sequence selected from: SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 or SEQ ID NO: 44, and (b) A heavy chain comprising an amino acid sequence selected from the following: SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 or SEQ ID NO: 52.

[0077] In some embodiments of the present invention, the monoclonal antibody includes: (i) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 45; or (ii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 36, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 46; or (iii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 37, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47; or (iv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47; or (v) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 48; or (vi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 49; or (vii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (viii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (ix) (a) A light chain containing the amino acid sequence of SEQ ID NO: 40, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (x) (a) A light chain containing the amino acid sequence of SEQ ID NO: 42, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (xi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 37, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xiii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 41, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52; or (xiv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 36, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xvi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 40, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xvii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 42, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xviii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (ixx) (a) A light chain containing the amino acid sequence of SEQ ID NO: 43, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52; or (xx) (a) A light chain containing the amino acid sequence of SEQ ID NO: 44, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52.

[0078] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is selected from the following antibodies: 04-002, 04-004, 04-009, 04-013, 04-016, 04-024, 04-042, 04-045, 04-046, 04-047, 03-008, 03-010, 03-015, 03-023, 03-024, 03-026, 03-027, 03-028, 03-036, or 03-038.

[0079] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-002.

[0080] Antibody 04-002 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 35; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 45.

[0081] Antibody 04-002 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 29.

[0082] Antibody 04-002 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 13, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0083] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-004.

[0084] Antibody 04-004 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 36; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 46.

[0085] Antibody 04-004 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 20; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 29.

[0086] Antibody 04-004 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 13, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0087] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-009.

[0088] Antibody 04-009 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 37; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47.

[0089] Antibody 04-009 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 21; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 30.

[0090] Antibody 04-009 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 14, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0091] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-013.

[0092] Antibody 04-013 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 38; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47.

[0093] Antibody 04-013 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 22; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 30.

[0094] Antibody 04-013 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 14, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0095] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-016.

[0096] Antibody 04-016 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 39; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 48.

[0097] Antibody 04-016 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 23; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 30.

[0098] Antibody 04-016 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 14, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0099] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-024.

[0100] Antibody 04-024 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 39; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 49.

[0101] Antibody 04-024 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 23; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 31.

[0102] Antibody 04-024 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 13, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0103] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-042.

[0104] Antibody 04-042 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 35; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50.

[0105] Antibody 04-042 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32.

[0106] Antibody 04-042 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 15, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0107] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-045.

[0108] Antibody 04-045 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 38; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50.

[0109] Antibody 04-045 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 22; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32.

[0110] Antibody 04-045 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 15, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0111] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-046.

[0112] Antibody 04-046 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 40; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50.

[0113] Antibody 04-046 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 24; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32.

[0114] Antibody 04-046 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 15, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0115] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 04-047.

[0116] Antibody 04-047 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 42; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50.

[0117] Antibody 04-047 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 26; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 32.

[0118] Antibody 04-047 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 15, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0119] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-008.

[0120] Antibody 03-008 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 37; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0121] Antibody 03-008 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 21; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0122] Antibody 03-008 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0123] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-010.

[0124] Antibody 03-010 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 35; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0125] Antibody 03-010 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 19; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0126] Antibody 03-010 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0127] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-015.

[0128] Antibody 03-015 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 41; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52.

[0129] Antibody 03-015 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 25; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34.

[0130] Antibody 03-015 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 3, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 7, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 10, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 17, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0131] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-023.

[0132] Antibody 03-023 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 36; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0133] Antibody 03-023 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 20; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0134] Antibody 03-023 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0135] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-024.

[0136] Antibody 03-024 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 38; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0137] Antibody 03-024 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 22; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0138] Antibody 03-024 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0139] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-026.

[0140] Antibody 03-026 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 40; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0141] Antibody 03-026 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 24; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0142] Antibody 03-026 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0143] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-027.

[0144] Antibody 03-027 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 42; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0145] Antibody 03-027 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 26; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0146] Antibody 03-027 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 11, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0147] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-028.

[0148] Antibody 03-028 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 39; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51.

[0149] Antibody 03-028 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 23; (b) A heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 33.

[0150] Antibody 03-028 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 8, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0151] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-036.

[0152] Antibody 03-036 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 43; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52.

[0153] Antibody 03-036 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 27; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34.

[0154] Antibody 03-036 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 10, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 17, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0155] In some embodiments of the present invention, the monoclonal antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is antibody 03-038.

[0156] Antibody 03-038 includes: (a) A light chain containing the amino acid sequence of SEQ ID NO: 44; and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52.

[0157] Antibody 03-038 includes: (a) A light chain variable domain containing the amino acid sequence of SEQ ID NO: 28; (b) Heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 34.

[0158] Antibody 03-038 includes: (a) Contains the following light chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 7, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 9, and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12, (ii) CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 17, (iii) CDR3 (Kabat) having the amino acid sequence of SEQ ID NO: 18.

[0159] The hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the light and heavy chain variable domains of all the above-described antibodies are provided according to the Kabat nomenclature. Those skilled in the art will understand that the hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the light and heavy chain variable domains can also be represented according to other commonly known numbering schemes, such as IMGT, Chothia, or AbM. Therefore, the present invention also covers all the above-described antibodies characterized by the hypervariable regions (LCDR1, 2, 3 and HCDR1, 2, 3) of the light and heavy chain variable domains using the IMGT, Chothia, or AbM numbering scheme.

[0160] The antibody according to the present invention specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, exhibiting a high binding affinity parameter for the TRBV5-1 segment of the variable domain of the human T cell receptor β chain. Furthermore, the antibody according to the present invention is humanized, stable and thermostable in human serum, and also possesses antibody-dependent cell-mediated cytotoxicity (ADCC).

[0161] Nucleic acid molecules In one aspect, the present invention relates to nucleic acids encoding any of the above-described antibodies or antigen-binding fragments thereof, said antibodies or antigen-binding fragments thereof specifically binding to the TRBV5-1 region of the variable domain of the human T cell receptor β chain.

[0162] In any of the described embodiments, the nucleic acid molecules can be isolated.

[0163] The terms “nucleic acid,” “nucleic acid sequence,” “nucleic acid sequence,” “polynucleotide,” “oligonucleotide,” “polynucleotide sequence,” and “nucleotide sequence,” which are used interchangeably in this specification, refer to the precise sequence of modified or unmodified nucleotides that determines a fragment or region of a nucleic acid, whether it contains non-natural nucleotides, and whether it is double-stranded DNA or RNA, single-stranded DNA or RNA, or a transcript of said DNA.

[0164] Unless otherwise indicated, the term nucleotide sequence encompasses its complement. Therefore, nucleic acid having a specific sequence should be understood to encompass nucleic acid having its complementary strand.

[0165] An "isolated" nucleic acid molecule is a molecule that is identified and separated from at least one nucleic acid molecule impurity. An isolated nucleic acid molecule differs from the form or set of molecules found under natural conditions. Therefore, an isolated nucleic acid molecule differs from the nucleic acid molecules that exist naturally in cells.

[0166] In one aspect, the present invention relates to nucleic acid molecules comprising a nucleotide sequence encoding an amino acid sequence selected from SEQ ID NO: 53-86. The nucleic acid molecule may also comprise any combination of said nucleotide sequences.

[0167] As those skilled in the art will understand, due to the redundancy of the genetic code, various different DNA sequences can encode the amino acid sequences of the light or heavy chains of antibodies or fragments thereof (VH, VL, CDR, etc.) according to the present invention. Generating these alternative DNA sequences encoding one and the same amino acid sequence is entirely within the skill of those specially trained in the art. Such variant DNA sequences are within the scope of the present invention.

[0168] In some embodiments of the present invention, nucleic acid is DNA.

[0169] The nucleic acid molecules of the present invention can be isolated from any source that produces monoclonal antibodies or antigen-binding fragments thereof, said antibodies or antigen-binding fragments specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain. In some embodiments of the present invention, the nucleic acid molecules of the present invention can be synthesized by chemical synthesis rather than isolation.

[0170] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-002, 04-042, and 03-010, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 53.

[0171] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-004 and 03-023, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 54.

[0172] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-009 and 03-008, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 55.

[0173] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-013, 04-045, and 03-024, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 56.

[0174] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-016, 04-024, and 03-028, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 57.

[0175] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 04-046 and 03-026, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 58.

[0176] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibody 03-015, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 59.

[0177] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibodies 03-027 and 04-047, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 60.

[0178] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibody 03-036, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 61.

[0179] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain variable domain of antibody 03-038, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 62.

[0180] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibodies 04-002 and 04-004, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 63.

[0181] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibodies 04-009, 04-013, and 04-016, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 64.

[0182] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibody 04-024, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 65.

[0183] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibodies 04-042, 04-045, 04-046, and 04-047, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 66.

[0184] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibodies 03-008, 03-010, 03-023, 03-024, 03-026, 03-027, and 03-028, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 67.

[0185] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain variable domain of antibodies 03-015, 03-036, and 03-038, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 68.

[0186] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-002, 04-042, and 03-010, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 69.

[0187] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-004 and 03-023, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 70.

[0188] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-009 and 03-008, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 71.

[0189] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-013, 04-045, and 03-024, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 72.

[0190] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-016, 04-024, and 03-028, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 73.

[0191] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 04-046 and 03-026, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 74.

[0192] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibody 03-015, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 75.

[0193] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibodies 03-027 and 04-047, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 76.

[0194] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibody 03-036, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 77.

[0195] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the light chain of antibody 03-038, and includes a nucleic acid having a nucleotide sequence having SEQ ID NO: 78.

[0196] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain of antibody 04-002, and includes a nucleic acid having a nucleotide sequence having SEQ ID NO: 79.

[0197] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain of antibody 04-004, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 80.

[0198] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain of antibodies 04-009 and 04-013, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 81.

[0199] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain of antibody 04-016, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 82.

[0200] In some embodiments of the present invention, the nucleic acid is the amino acid sequence encoding the heavy chain of antibody 04-024, and includes the nucleic acid having the nucleotide sequence of SEQ ID NO: 83.

[0201] In some embodiments of the present invention, the nucleic acid is the amino acid sequence encoding the heavy chain of antibodies 04-042, 04-045, 04-046, and 04-047, and includes the nucleic acid having the nucleotide sequence of SEQ ID NO: 84.

[0202] In some embodiments of the present invention, the nucleic acid is an amino acid sequence encoding the heavy chain of antibodies 03-008, 03-010, 03-023, 03-024, 03-026, 03-027, and 03-028, and includes a nucleic acid having the nucleotide sequence of SEQ ID NO: 85.

[0203] In some embodiments of the present invention, the nucleic acid is the amino acid sequence encoding the heavy chain of antibodies 03-015, 03-036, and 03-038, and includes the nucleic acid having the nucleotide sequence of SEQ ID NO: 86.

[0204] Nucleic acid molecules can be used to express recombinant monoclonal antibodies or their antigen-binding fragments, which specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0205] carrier In one aspect, the present invention relates to expression vectors comprising any of the aforementioned nucleic acid molecules, said nucleic acid molecules encoding an antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, or a corresponding amino acid sequence (e.g., heavy chain and / or light chain binding domain sequence) of said portion. The present invention relates to vectors suitable for expressing any of the nucleotide sequences described herein.

[0206] As used in this article, the term "carrier" refers to a nucleic acid molecule that can transport another nucleic acid to which it has been linked.

[0207] As used in this specification, the term "expression" is defined as the transcription and / or translation of a specific nucleotide sequence driven by its promoter.

[0208] In some embodiments of the present invention, the vector is a plasmid, i.e., a circular double-stranded DNA fragment into which additional DNA segments can be inserted.

[0209] In some embodiments of the present invention, the vector is a viral (expression) vector, wherein additional DNA segments may be inserted into the viral genome.

[0210] In some embodiments of the invention, vectors are capable of autonomous replication in the host cells to which they are introduced (e.g., bacterial vectors and augmentation vectors with a bacterial replication initiation site). In further embodiments of the invention, vectors (e.g., non-augmentation vectors) can integrate into the host cell's genome after introduction into the host cell and thus replicate along with the host genome. Furthermore, some vectors are capable of directing the expression of genes operatively linked to them. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors").

[0211] In some embodiments of the present invention, the expression vector includes plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAVs), plant viruses (e.g., cauliflower mosaic virus, tobacco mosaic virus), granules, YACs, etc. DNA molecules can be inserted into the vector, allowing the transcriptional and translational control sequences within the vector to perform their intended functions of regulating DNA transcription and translation. The expression vector and expression control sequences can be selected to be compatible with the host cells used for expression.

[0212] In one embodiment of the invention, DNA molecules encoding partial or complete heavy and light chain sequences can be inserted into different vectors.

[0213] In one implementation, any combination of the above-described DNA molecules is introduced into the same expression vector.

[0214] In one embodiment of the invention, DNA molecules can be introduced into an expression vector by standard methods (e.g., ligation of a gene fragment of an antibody to a complementary restriction site on a vector, or blunt-end ligation if no restriction site is present).

[0215] In some embodiments of the invention, a suitable vector is one that includes a restriction site, allowing for the easy insertion and expression of any VH or VL sequence, as described above. The recombinant expression vector may also encode a signal peptide that promotes the secretion of antibody chains from host cells. The antibody chain gene can be cloned into the vector such that the signal peptide is linked in-frame to the N-terminus of an immunoglobulin chain. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide derived from a non-immunoglobulin).

[0216] In some embodiments of the invention, the vector may include expression control sequences. As used herein, the term "expression control sequence" refers to a polynucleotide sequence necessary for the expression and processing of the coding sequence into which it is inserted. Those skilled in the art will understand that the design of expression vectors, including the selection of expression control sequences, may depend on factors such as the type of host cell to be transformed, the desired antibody expression level, and so on. Expression control sequences include appropriate transcription initiation, termination, promoter, and enhancer sequences; effective RNA processing signals such as splicing and polyadenylation signals; sequences stabilizing cytoplasmic mRNA; sequences enhancing translation efficiency (i.e., Kozak concordant sequences); sequences enhancing protein stability; and sequences enhancing protein secretion when necessary. The nature of such expression control sequences varies depending on the host organism; in prokaryotes, such expression control sequences typically include a promoter, a ribosome binding site, and a transcription termination sequence; in eukaryotes, such expression control sequences typically include a promoter and a transcription termination sequence. Preferred expression control sequences for mammalian host cells include viral elements that ensure high levels of protein expression in mammalian cells, such as those derived from promoters and / or enhancers of: retroviral LTRs (long terminal repeats), cytomegaloviruses (CMV) (e.g., CMV enhancers / promoters), simian virus 40 (SV40) (e.g., SV40 promoters / enhancers), adenoviruses (e.g., major late promoter adenovirus (AdMLP)), polyomaviruses, and strong mammalian promoters (e.g., TTR promoters, innate immunoglobulin promoters, or actin promoters). The expression control sequence should at least cover all components whose presence is important for expression and processing.

[0217] In some embodiments of the present invention, in addition to the antibody chain gene and expression control sequence, the recombinant expression vector of the present invention may also carry other sequences, such as sequences that regulate vector replication in host cells (e.g., origin of replication) and selectable marker genes. Selectable marker genes facilitate selection of host cells into which the vector has been introduced.

[0218] host cells In one aspect, the present invention relates to a method for generating host cells to produce any of the above-described antibodies or antigen-binding fragments thereof, said antibodies or antigen-binding fragments thereof specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, and includes transforming cells with the above-described vector.

[0219] In one aspect, the present invention relates to a host cell that produces any of the above-described antibodies or antigen-binding fragments thereof, said antibody or antigen-binding fragment specifically binding to the TRBV5-1 region of the variable domain of the human T cell receptor β chain, said host cell comprising any of the above-described nucleic acids.

[0220] As used herein, the term "host cell" refers to a cell in which a recombinant expression vector has been introduced. This invention relates to host cells, which may include, for example, the vectors described above according to the invention. The invention further relates to host cells comprising, for example, a nucleotide sequence encoding a heavy chain or its antigen-binding portion, a nucleotide sequence encoding a light chain or its antigen-binding portion, or both. It should be understood that "host cell" refers not only to the specific subject cell but also to the progeny of such cells. Because modifications may occur in offspring due to mutations or environmental influences, such progeny may not actually be identical to the parent cells; however, such cells are still included within the scope of the term "host cell" as used herein.

[0221] Nucleic acid molecules encoding monoclonal antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 region of the variable domain of the human T cell receptor β chain according to the present invention, and vectors containing such nucleic acid molecules, can be used for transfection of mammalian cells, plant cells, bacterial cells, or yeast cells. Transfection can be performed by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, cationic polymer-nucleic acid complex transfection, transfection by electroporation, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, encapsulation of polynucleotides into liposomes, and direct microinjection of DNA into the nucleus. Alternatively, nucleic acid molecules can also be introduced into mammalian cells via viral (expression) vectors.

[0222] Mammalian cell lines used as hosts for transformation are known in the art and include a variety of immortalized cell lines available. These include, for example, Chinese hamster ovary (CHO) cells, NS0 cells, SP2 cells, HEK-293T cells, FreeStyle 293 cells (Invitrogen), NIH-3T3 cells, HeLa cells, young hamster kidney (BHK) cells, African green monkey kidney cells (COS) cells, human hepatocellular carcinoma cells (e.g., Hep G2), A549, SK-HEP1, HUH7, Hep-RG cells, and many other cell lines. Cell lines are selected by identifying which cell lines have high expression levels and provide the necessary characteristics for the protein to be produced. Other cell lines that can be used are insect cell lines, such as Sf9 or Sf21 cells. When a recombinant expression vector encoding a monoclonal antibody or its antigen-binding fragment that specifically binds to the TRBV5-1 region of the variable domain of the human T-cell receptor β-chain is introduced into mammalian host cells, the antibody or its fragment is produced by culturing the host cells for a period of time sufficient to allow expression of the antibody or its fragment in the host cells, or more preferably, the antibody or its fragment is secreted into the culture medium in which the host cells are grown. The monoclonal antibody or its antigen-binding fragment that specifically binds to the TRBV5-1 region of the variable domain of the human T-cell receptor β-chain can be isolated from the culture medium using standard protein purification techniques. Plant host cells include, for example, *Nicotiana*, *Arabidopsis*, duckweed, maize, wheat, and potato. Bacterial host cells include species of *Escherichia* and *Streptomyces*. Yeast host cells include *Schizosaccharomyces pombe*, *Saccharomyces cerevisiae*, and *Pichia pastoris*.

[0223] Furthermore, various known techniques can be used to enhance the production levels of monoclonal antibodies or antigen-binding fragments thereof from production cell lines that specifically bind to the TRBV5-1 region of the variable domain of the human T cell receptor β chain. For example, the glutamine synthase gene expression system (GS system) is a common method used to enhance expression under certain conditions.

[0224] Monoclonal antibodies or antigen-binding fragments thereof derived from various cell lines will likely have glycosylation profiles that differ from each other, and these monoclonal antibodies or antigen-binding fragments specifically bind to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain. However, regardless of the glycosylation of the binding molecule, and generally regardless of the presence or absence of post-translational modifications, monoclonal antibodies or antigen-binding fragments thereof encoded by the nucleic acid molecules described herein or containing the amino acid sequences provided herein are part of this invention, and these monoclonal antibodies or antigen-binding fragments specifically bind to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain.

[0225] The aforementioned host cells do not involve the use of host cells derived from human embryos.

[0226] The aforementioned host cells do not refer to host cells generated by modifying the genetic integrity of human germ cells.

[0227] Methods for producing antibodies In one aspect, the present invention relates to a method for producing an antibody or an antigen-binding fragment thereof, said antibody or antigen-binding fragment thereof specifically binding to the TRBV5-1 segment of the variable domain of the β chain of a human T cell receptor, said method comprising culturing said host cells in a growth medium under conditions sufficient to produce said antibody or fragment thereof, followed by isolation and purification of the resulting antibody or fragment thereof.

[0228] Pharmaceutical Composition Another aspect of the invention is a pharmaceutical composition comprising a monoclonal antibody or an antigen-binding fragment thereof according to the invention as an active ingredient (or as the sole active ingredient), said monoclonal antibody or antigen-binding fragment thereof specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0229] In one aspect, the present invention relates to pharmaceutical compositions comprising any of the aforementioned antibodies or antigen-binding fragments thereof in combination with one or more pharmaceutically acceptable excipients.

[0230] In one aspect, the present invention relates to pharmaceutical compositions for treating or preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising any of the aforementioned antibodies or antigen-binding fragments thereof in combination with one or more pharmaceutically acceptable excipients.

[0231] In one aspect, the present invention relates to pharmaceutical compositions for treating or preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising, in combination with one or more pharmaceutically acceptable excipients, a therapeutically effective amount of any of the aforementioned antibodies or antigen-binding fragments thereof. "Pharmaceutical composition" means a composition comprising an antibody according to the invention and at least one component selected from: pharmaceutically acceptable and pharmacologically compatible fillers, solvents, diluents, carriers, excipients, distributors and sensors, and delivery agents.

[0232] The term “pharmaceutical acceptable” refers to one or more compatible liquid or solid components that are suitable for administration in mammals, preferably humans.

[0233] The term "excipient" is used herein to describe any component other than the antibody according to the invention. These are substances with inorganic or organic properties that are used in the production / manufacturing of pharmaceuticals to impart the necessary physicochemical properties to the pharmaceutical product.

[0234] In some embodiments, the composition is intended to improve, prevent or treat diseases or conditions that can be mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0235] The term “disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain” refers to all diseases or conditions directly or indirectly related to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, including the etiology, development, progression, persistence, or pathological condition of the disease or condition.

[0236] "Treatment" and "treatment" refer to methods for relieving or eliminating a biological condition and / or at least one of its accompanying symptoms. Furthermore, this article uses the term "treatment" to include curative treatment, palliative treatment, and preventative treatment.

[0237] The "prevention" according to the present invention is a series of measures aimed at preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0238] The term "symptom" means any condition that would benefit from treatment or prevention according to the present invention. The definition of this term includes both chronic and acute symptoms or diseases, including those pathological conditions that make mammals susceptible to the symptoms discussed.

[0239] "Therapeutic effective dose" refers to the amount of therapeutic agent to be administered during treatment or prevention that, to a certain extent, alleviates one or more symptoms of the disease for which it provides treatment or prevention. Therapeutic effective dose can vary depending on factors such as the specific condition for which it provides treatment or prevention, the patient's age, sex, and weight, and whether the monoclonal antibody or its antigen-binding fragment specifically binds to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain is administered as a single treatment or prevention, and / or in combination with one or more other drugs, treatments, or prevention methods.

[0240] In one respect, the subject or patient for treatment or prevention is a human subject. The subject can be male or female of any age.

[0241] The pharmaceutical compositions and their preparation methods of the present invention are readily apparent to those skilled in the art. The pharmaceutical compositions should preferably be manufactured in accordance with GMP (Good Manufacturing Practice) requirements.

[0242] In some embodiments of the pharmaceutical composition, it may include a buffer composition, a tensile agent (permeasurant or permeasurant), a stabilizer, and / or a solubilizer.

[0243] The pharmaceutical composition according to the present invention is a stable composition.

[0244] A pharmaceutical composition is "stable" if the active agent retains its physical and / or chemical stability and / or biological activity at a storage temperature of, for example, 2-8°C, during a specified shelf life. Preferably, the active agent retains both physical and chemical stability, as well as biological activity. The shelf life is adjusted based on the results of stability tests under accelerated or natural aging conditions.

[0245] In some implementations, the pharmaceutical composition is an injectable dosage form.

[0246] In some implementations, the injectable dosage form is an infusion solution.

[0247] In some implementations, the injectable dosage form is a solution for subcutaneous administration.

[0248] Injectable dosage forms can be manufactured without restriction in unit dosage forms, such as in ampoules, vials, plastic containers, pre-filled syringes, or automated injection devices.

[0249] In some embodiments, the pharmaceutical composition is provided in a dry form (i.e., powder or granule form) for reconstitution with a suitable solvent (e.g., sterile, pyrogen-free water) prior to administration. Such pharmaceutical formulations can be prepared, for example, by lyophilization, which is referred to in the art as freeze-drying and involves a process of freezing the product and subsequently removing the solvent from the frozen material.

[0250] In some embodiments, the pharmaceutical composition is a lyophilized product used to prepare a solution for infusion.

[0251] In some embodiments, the pharmaceutical composition is a lyophilized product used to prepare a solution for subcutaneous administration.

[0252] In some embodiments, the pharmaceutical composition is a concentrate used to prepare a solution for infusion.

[0253] In some embodiments, the pharmaceutical composition is a concentrate used to prepare a solution for subcutaneous administration.

[0254] In one aspect, the present invention relates to pharmaceutical compositions comprising a monoclonal antibody or antigen-binding fragment thereof according to the invention that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, and at least one other therapeutically active compound.

[0255] In one aspect, the present invention relates to pharmaceutical compositions for treating or preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising any of the aforementioned antibodies or antigen-binding fragments thereof, and at least one other therapeutically active compound.

[0256] In one aspect, the present invention relates to pharmaceutical compositions comprising any of the aforementioned antibodies or antigen-binding fragments thereof, and at least one further therapeutically active compound.

[0257] In one aspect, the present invention relates to pharmaceutical compositions for treating or preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising any of the aforementioned antibodies or antigen-binding fragments thereof, and at least one further therapeutically active compound.

[0258] In one aspect, the present invention relates to pharmaceutical compositions for treating or preventing diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising any of the aforementioned antibodies or antigen-binding fragments thereof, and at least one other therapeutically active compound, which is an antibody, an insulin preparation, a small molecule, a hormone therapy agent, or any combination thereof.

[0259] In some embodiments of the pharmaceutical composition, other therapeutically active compounds are selected from the following: insulin preparations, glucocorticoids, β2-adrenergic receptor agonists, M-type cholinergic receptor antagonists, budesonide, azathioprine, methotrexate, dapsone, or any combination thereof.

[0260] In some embodiments of the pharmaceutical composition, the disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain is selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

[0261] Monoclonal antibodies or their antigens that specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain Therapeutic uses of combined fragments In one respect, an antibody or antigen-binding fragment thereof that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is used to treat or prevent diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

[0262] In one respect, the subject or patient for treatment or prevention is a human subject. The subject can be male or female of any age.

[0263] In one aspect, the present invention relates to a method for treating or preventing a disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising administering to a subject requiring such treatment or prevention a therapeutically effective amount of any of the aforementioned antibodies or antigen-binding fragments thereof, or the aforementioned pharmaceutical compositions.

[0264] In one aspect, the present invention relates to a method for treating or preventing a disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising administering to a subject requiring such treatment or prevention a therapeutically effective amount of any of the aforementioned antibodies or antigen-binding fragments thereof, and at least one other therapeutically active compound.

[0265] In some implementations of treatment or prevention methods, diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain are selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

[0266] In some implementations of treatment or prevention methods, other therapeutically active compounds are antibodies, insulin preparations, small molecules, hormone therapy agents, or combinations thereof.

[0267] In one aspect, the present invention relates to the use of the aforementioned antibody or its antigen-binding fragment or the aforementioned pharmaceutical composition for the treatment or prevention of diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain in subjects requiring such treatment or prevention.

[0268] In one aspect, the present invention relates to the use of the aforementioned antibody or its antigen-binding fragment or the aforementioned pharmaceutical composition, as well as at least one other therapeutically active compound, for the treatment or prevention of a disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain in a subject requiring such treatment or prevention.

[0269] In some embodiments of the use, the disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain is selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

[0270] In some embodiments of the application, other therapeutically active compounds are antibodies, insulin preparations, small molecules, hormone therapy agents, or combinations thereof.

[0271] The use or method of using antibodies or their antigen-binding fragments (which specifically bind to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain) together with one or more other therapeutic agents is intended to refer to, and includes, the following: 1) When such components are formulated together into a single dosage form for administration to the patient at substantially the same time, such combinations of an antibody or antigen-binding fragment thereof specifically binding to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain and a therapeutic agent are administered simultaneously to patients requiring treatment or prophylaxis. 2) When such components are formulated separately into separate dosage forms and administered to the patient at substantially the same time, and subsequently released to the patient at substantially the same time, such combinations of an antibody or antigen-binding fragment thereof specifically binding to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain and a therapeutic agent are simultaneously administered to a patient requiring treatment or prophylaxis. 3) When such components are formulated separately into separate dosage forms for continuous administration by the patient, with significant time intervals between each administration, and subsequently released to the patient at substantially different times, such combinations of antibodies or antigen-binding fragments specifically binding to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain and therapeutic agents are sequentially administered to patients requiring treatment or prophylaxis; and 4) When such components are formulated together into a single dosage form for controlled release of said components, and subsequently released simultaneously, sequentially or jointly to said patient at the same time and / or at different times, wherein each part can be administered via the same or different routes, such combinations of an antibody or its antigen-binding fragment specifically binding to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain and a therapeutic agent are sequentially administered to patients requiring treatment or prevention.

[0272] Antibodies or their antigen-binding fragments that specifically bind to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain can be administered as a stand-alone therapy without further therapeutic treatment or prophylaxis.

[0273] In some embodiments of a treatment or preventative method or use, an antibody or antigen-binding fragment thereof that specifically binds to the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain may be administered in combination with other therapeutically active compounds selected from: insulin preparations, glucocorticoids, β2-adrenergic receptor agonists, M-type cholinergic receptor antagonists, budesonide, azathioprine, methotrexate, dapsone, or any combination thereof.

[0274] In some implementations, the treatment or preventive method or use requires selection from the following pretreatments: histamine receptor blockers, glucocorticoids, antiemetics, acetaminophen, or any combination thereof.

[0275] The antibodies or antigen-binding fragments thereof that specifically bind to the TRBV5-1 region according to the present invention, as well as pharmaceutical compositions, are suitable for parenteral administration in the form of sterile pharmaceutical products, intended to be administered to the subject by injection, infusion, or implantation bypassing the gastrointestinal tract by disrupting the integrity of the skin or mucous membranes. In particular, parenteral administration includes subcutaneous, intraperitoneal, intramuscular, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, ​​intracranial, intrasynovial, percutaneous injection or infusion; and renal dialysis infusion techniques.

[0276] In some embodiments of treatment or prevention methods or uses, an antibody or antigen-binding fragment or pharmaceutical composition that specifically binds to the TRBV5-1 segment is administered intravenously.

[0277] In some implementations, intravenous administration is performed by infusion, prolonged infusion, or long-acting continuous infusion.

[0278] In some embodiments of treatment or prevention methods or uses, an antibody or antigen-binding fragment or pharmaceutical composition that specifically binds to the TRBV5-1 segment is administered subcutaneously.

[0279] In some implementations, subcutaneous administration is performed by subcutaneous injection.

[0280] In some embodiments of treatment or prevention methods or uses, a suitable dose of the monoclonal antibody or its antigen-binding fragment that specifically binds to the TRBV5-1 segment according to the present invention is in the range of 0.1–200 mg / kg. Brief description of the attached diagram Figure 1 It is a map of a vector carrying the genetic sequence of the light chain of the anti-TRBV5-1 antibody.

[0282] Figure 2 It is a map of a vector carrying the genetic sequence of the anti-TRBV5-1 antibody heavy chain.

[0283] about Figure 1 -2 .

[0284] Figure 3 This is an electrophoresis image of an antibody in 4-15% PAAG that does not contain additional disulfide bonds between the heavy and light chains (under reducing conditions).

[0285] 1) Protein molecular weight markers; corresponding to the molecular weight (kDa) of the lane; 2) 04-002; 3) 04-004; 4) 04-009; 5) 04-013; 6) 04-016; 7) 04-024; 8) 04-042; 9) 04-045; 10) 04-046; 11) 04-047.

[0286] Figure 4 This is an electrophoretic image of an antibody containing additional disulfide bonds between the heavy and light chains (under non-reducing conditions) in 4-15% PAAG.

[0287] 1) Protein molecular weight markers; corresponding to the molecular weight (kDa) of the lane; 2) 04-002; 3) 04-004; 4) 04-009; 5) 04-013; 6) 04-016; 7) 04-024; 8) 04-042; 9) 04-045; 10) 04-046; 11) 04-047.

[0288] Figure 5 The graph shows that antibodies 04-002 and 03-010 failed to induce antibody-dependent phagocytosis (ADCP) in the reporter cell assay.

[0289] Figure 6 The graph shows the non-specific activation of the NFAT signaling cascade by antibodies 04-002, 04-016, and 04-024 in the reporter cell assay.

[0290] Figure 7 The graph shows the non-specific activation of the NFAT signaling cascade by antibodies 03-010 and 03-015 in the reporter cell assay.

[0291] Figure 8 The graph shows the ability of antibodies 04-002, 04-016, 04-024, and 03-010 to induce antibody-dependent cell-mediated cytotoxicity (ADCC) in a suspension of peripheral blood mononuclear cells (PBMCs).

[0292] Figure 9This is a histogram showing the changes in the relative expression level of the TRBV5-1 gene in human peripheral blood mononuclear cells (PBMCs) in the presence of anti-TRBV5-1 antibody.

[0293] Figure 10 The graph shows the complement-free cytotoxicity (CDC) curves of antibodies 04-002 and 03-010.

[0294] Figure 11 This is a graph showing the ability of antibody 04-002 to not deplete the NK cell population in human PBMCs.

[0295] Figure 12 This is a graph showing the ability of antibody 04-002 to not deplete the B cell population in human PBMCs.

[0296] Figure 13 This is a graph showing the ability of antibody 04-002 to not deplete the large population of T cells in human PBMCs.

[0297] Figure 14 This is a graph showing the ability of antibody 04-002 to not deplete the mononuclear cell population in human PBMCs.

[0298] Figure 15 It is a graph showing the percentage of TRBV5-1 positive cells in human CD45 and CD3 positive cell populations and in mouse blood samples in the presence of anti-TRBV5-1 antibody at day 18.

[0299] ** - p-value ≤ 0.01 Statistical analysis was performed using the Kruskal-Wallis test and the Dunn test for multiple comparisons.

[0300] Figure 16 It is a graph showing the percentage of TRBV5-1 positive cells in human CD45 and CD3 positive cell populations and in mouse spleen samples at day 18.

[0301] *** - p-value ≤ 0.001 Statistical analysis was performed using the Kruskal-Wallis test and the Dunn test for multiple comparisons. Example

[0302] The following embodiments are provided to aid in a better understanding of the invention. These embodiments are for illustrative purposes only and should not be construed as limiting the scope of the invention in any way.

[0303] Materials and General Methods General information about the nucleotide sequences of the light and heavy chains of human immunoglobulins is given in the following: Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991). The amino acids of the antibody chains are numbered according to the EU numbering scheme (Edelman, G.M., et al., Proc. Natl. Acad. Sci. USA 63 (1969) 78-85; Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).

[0304] Recombinant DNA technology Standard methods for manipulating DNA are described in Sambrook, J. et al., Molecular cloning: A laboratory manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989. Molecular biology reagents are used according to the manufacturer's instructions.

[0305] Gene synthesis The required gene segments were prepared from oligonucleotides synthesized chemically. Each segment, flanked by a single restriction site and 300-1400 bp in length, was assembled via oligonucleotide annealing and ligation, including PCR amplification, and subsequently cloned through the restriction site. The DNA sequence of the subcloned gene fragment was confirmed by DNA sequencing.

[0306] DNA sequencing The DNA sequence was determined using Sanger sequencing.

[0307] DNA and protein sequence analysis and sequence data management Unipro's UGENE suite version 1.29 and SnapGene version 6.1 are used for sequence generation, plotting, analysis, annotation, and visualization.

[0308] expression carrier To express the antibody described in the application materials, a variant of the expression plasmid intended for antibody expression in prokaryotic cells (Escherichia coli) and transient expression in eukaryotic cells (e.g., CHO cells) is used. In addition to the antibody expression cassette, the vector also contains: a replication origin allowing the plasmid to replicate in E. coli, and a gene conferring resistance to various antibiotics (e.g., ampicillin, kanamycin) in E. coli.

[0309] The fusion gene containing the subject antibody chain, as described below, is generated via PCR and / or gene synthesis, and assembled using known recombination methods and techniques, for example, by ligating the corresponding nucleic acid segments using unique restriction sites in the appropriate vector. The subcloned nucleic acid sequence is verified by DNA sequencing. For transient transfection, a larger number of plasmids are prepared using plasmid preparation from transformed *E. coli* cultures.

[0310] Example 1. Selection of anti-TRBV5-1 antibody sequence.

[0311] Anti-TRBV5-1 antibody molecules are used on a computer chip ( in silico The structural data obtained is used to generate the scaffold. The scaffold construction on the computer chip is performed using JSC's internal algorithm "Biocad". A PrepWizard instrument from Schrodinger Suite 2021-3 is used to fabricate the structure. Subsequently, folding is performed using the Prime tool from Schrodinger 2021-3.

[0312] A method for sequential modification of amino acid composition using variable structural domains.

[0313] The amino acid sequence of the antibody was optimized on a computer chip to produce the antibody shown in Table 1.

[0314] Table 1. Anti-TRBV5-1 antibody.

[0315] Example 2. Humanization analysis of heavy and light chain variable fragments of anti-TRBV5-1 antibody.

[0316] Table 2 shows the results of humanization analysis of the heavy and light chain variable fragments of the anti-TRBV5-1 antibody compared with the control product 01-010, whose sequence is known according to international application WO2020142672.

[0317] Table 2. Humanization degree of variable fragments in antibodies.

[0318] Therefore, the heavy and light chain variable fragments of the anti-TRBV5-1 antibody developed according to the present invention have a humanization degree of more than 85%.

[0319] Example 3: Obtaining the anti-TRBV5-1 antibody sequence.

[0320] The genes for the heavy and light chain variable domains of the anti-TRBV5-1 antibody according to the present invention, selected from Table 1, were assembled by de novo synthesis of oligonucleotides. Nucleotide sequences were selected based on amino acid sequences; codon composition was optimized for CHO expression cells. For each variable fragment sequence, partially complementary primers with 20 bp overlap were selected, and PCR amplification was performed. The resulting fragments were combined by PCR using the CK constant domain sequence for the light chain variable domain and the CH1-CH2-CH3 heavy chain constant domain sequence for the heavy chain variable domain, and cloned as a portion of the pintA expression vector. Figure 1-2 The plasmids were assembled using restriction-ligation or ligase-free cloning methods. The nucleotide sequence of the gene was confirmed by DNA sequencing according to the Sanger method. The plasmid DNA preparation was purified from E. coli cell cultures using a commercial kit from Qiagen. The resulting DNA was used for transient protein production in CHO cell lines.

[0321] Example 4: Modification of the Fc heavy chain constant domain of anti-TRBV5-1 antibodies 04-002, 04-016, and 04-024.

[0322] To generate antibodies with improved properties, the heavy chain constant domain is modified by deleting the C-terminal amino acids glycine and lysine (delGK). This modification allows for increased uniformity in protein formulations.

[0323] The genetic construct was assembled as follows: the heavy chain variable fragment sequences of antibodies 04-002, 04-016, and 04-024 were cloned as part of the pintA vector and combined with the CH1-CH2-CH3 heavy chain constant domain sequence modified with delGK. Cloning was performed using a restriction-ligase method, and the resulting sequences were confirmed by Sanger sequencing.

[0324] Plasmid DNA preparations were produced by purifying E. coli cell cultures using a commercial kit from Qiagen. born. The obtained DNA was used to transiently produce proteins in CHO cell lines.

[0325] Example 5: Production of antibody preparations and determination of antibody functional activity.

[0326] To generate antibodies, we cultured transfected CHO cells for 24 hours at a fucose analog density of 1.8–2.2 million / ml with plasmid DNA complexed with polyethyleneimine. One day post-transfection, cells were cultured at a lower temperature with cell growth supplements. 4–7 days post-transfection, antibody titers were measured using OctetRed96 (Pall) according to the manufacturer’s standard guidelines for the proA biosensor; the culture supernatant containing the subject antibody was then diluted to 3–10 μg / ml and incubated with J.RT3-T3.5-TRBV5-1 cells. Cell fluorescence was measured by flow cytometry after incubation with fluorescently labeled a-hFc-PE secondary antibody (Jackson ImmunoResearch) to confirm the binding of the anti-TRBV5-1 antibody to the TRBV5-1 antigen, indicating the antibody’s functional activity. J.RT3-T3.5-TRBV5-1 cells stained with secondary antibody alone were used as negative binding controls; J.RT3-T3.5-TRBV5-1 cells stained with commercial anti-TRBV5-1 antibody (Beckman Coulter IM2285) were used as positive controls.

[0327] Table 3 shows that the 20 anti-TRBV5-1 antibodies produced bind to the TRBV5-1 antigen, which is partially expressed as a multi-protein TRC-CD3 complex on the cell surface.

[0328] Table 3: Concentration and functional activity of anti-TRBV5-1 antibodies generated in a transient expression system based on Chinese hamster CHO cells.

[0329] Example 6. Antibody purification from suspension cultures of mammalian cells.

[0330] The antibody was purified from CHO cell growth medium. Seven days after production, the cell culture medium was clarified by centrifugation at 8000g followed by filtration through a 0.22 µm filter. Proteins were separated using a 5 ml gravity chromatography column from Thermo Scientific, packed with 0.5–1 ml of protein A adsorbent. The clarified culture medium was passed through the column, the adsorbent was washed with phosphate-buffered saline (PBS) at pH 7.4, and the antibody was eluted with 0.1 M glycine buffer at pH 3.0. The antibody eluent was adjusted to neutral using 1 M Tris-HCl buffer (pH 8.0). The antibody was then transferred to PBS (pH 7.4) by dialysis, concentrated if necessary, and purified using gel filtration on a Superdex 200 Increase 10 / 300 GL column equilibrated with PBS. The target peak was collected, filtered through a 0.22 µm filter, and stored at -80°C. The purity of the obtained protein solution was determined by SDS gel electrophoresis. Figure 3-4 Evaluation was conducted using HPLC and analytical HPLC.

[0331] Example 7. Measurement of the affinity constant for the interaction between the anti-TRBV5-1 antibody and the target antigen (TRBV5-1 segment).

[0332] Experiments were conducted on antibody affinity for the human TRAV1-2 / TRBV5-1 antigen using a ForteBio Octet RED 384 instrument with a protein A sensor (ForteBio). Analysis was performed at 30°C using a kinetic buffer (4.3 mM Na₂HPO₄; 136.9 mM NaCl, 1.5 mM KH₂PO₄; 2.7 mM KCl; 0.1% Tween 20 by volume; 0.1% BSA by mass; pH 7.4). Prior to measurement, the sensor was regenerated using a 10 mM glycine solution supplemented with HCl (pH 2.0). The regeneration process consisted of three repetitions of the following steps: 5 seconds in the regeneration solution and 5 seconds in the kinetic buffer for neutralization. The antibody was immobilized on the sensor surface in the kinetic buffer for 300 seconds. The concentration of the antibody to be immobilized was 10 μg / ml. After immobilization, a baseline was recorded in the kinetic buffer (120 seconds). During the binding phase (duration - 60 seconds), the sensor with the antibody-loaded antibody was immersed in wells containing a solution of the analyte (TRAV1-2 / TRBV5-1 antigen) prepared in the kinetic buffer. For each test antibody, a baseline of 10 μg / ml was recorded. Sensing plots were generated for analyte concentrations of 2.5 μg / ml (217.4 nM) and 2.5 μg / ml (54.4 nM). A kinetic buffer solution lacking the antigen was used as a reference signal (reference sensing plot). Dissociation of the antibody-antigen complex was recorded for 300 seconds after the binding phase. During the dissociation phase, the sensor was immersed in a well containing the kinetic buffer solution to record a baseline. To validate non-specific interactions between the analyte and the sensor (negative control), we used a sensor without antibody loading. During the loading step, the negative control sensor was immersed in a sodium acetate buffer solution without antibody, and all other steps were similar to those used for the antibody-loaded sensor.

[0333] The binding curves after subtracting the reference signal were analyzed using Octet Data Analysis software. A 1:1 interaction model was used for fitting with two sensor plots. The affinity of the subject antibody ranged from 8.4 to 69.4 nM.

[0334] Table 4. Measurement of the kinetic constants of the interaction between the antibody and the TRAV1-2 / TRBV5-1 antigen.

[0335] Conclusion: The anti-TRBV5-1 antibody according to the present invention specifically binds to the human TRBV5-1 antigen. The affinity of the obtained antibody exceeds that of antibody 01-010.

[0336] Example 8: Affinity constant analysis of the interaction between anti-TRBV5-1 antibody and Fcγ and FcRn receptors.

[0337] Measurements were performed according to the procedures outlined by the manufacturer of the streptavidin sensor (SA Biosensors) and the OctetRed96 (Pall) user manual. The sensor was prepared in the same manner as the affinity constant analysis for the target antigen.

[0338] Next, the sensor binds to the corresponding biotinylated receptor in 1x kinetic buffer, then is transferred to anti-TRBV5-1 antibody solutions at different concentrations in kinetic buffer, and finally, the sensor is immersed in a well containing kinetic buffer.

[0339] The obtained data were finally processed using a 1:1 or 2:1 model in Octet Data Analysis 8.2 software. The obtained affinity constants are shown in Table 5.

[0340] Table 5: Affinity constants for the interaction between anti-TRBV5-1 antibody 04-002 and Fcγ and FcRn receptors. Data were obtained using an Octet Red96 instrument, and constants were calculated using Octet Data Analysis 8.2 software.

[0341] Conclusion: Antibody 04-002 exhibits high affinity for the FcyR IIIa158V and IIIa158F receptors, potentially improving the effector properties of the subject antibody.

[0342] Example 9: Stability of anti-TRBV5-1 antibody during storage in human serum.

[0343] In this study, stability was understood to refer to the in vitro preservation of full-length antibody molecules in human serum at +37°C. To determine stability, samples of the tested antibodies were diluted in human serum to a concentration of 25 μg / ml and stored in a thermostat at +37°C for 14 days in a micro-volume of 150 μl supplemented with up to 0.1% sodium thimerosal. Unsupplemented human serum samples used for subsequent determination of background concentration values ​​for the samples in question were stored under similar conditions. After storage, antibody concentrations in the samples were determined by enzyme-linked immunosorbent assay (ELISA) with antibody calibration titrations ranging from 250 ng / ml to 3.9 ng / ml. Calibration and reference samples were diluted to the required concentrations immediately prior to the experiment.

[0344] For ELISA, the antigen containing the target human TRBV5-1 region was adsorbed onto the plate at a concentration of 2.5 μg / ml in 0.1M carbonate buffer, pH 9.5. The plate was washed, and the wells were blocked for 1 hour with 0.5% milk powder solution in wash buffer. 0.1% Tween-20 in PBS was used as the wash buffer. After blocking, the test antibody solution and calibration solution were added to the plate; all samples were diluted in the blocking solution and incubated at 37°C for 1 hour on a plate shaker. The plate was washed, and horseradish peroxidase-conjugated goat antibody against the human Ig Fc moiety (a-hFc-HRP) was added to the wells, and the plate was incubated at 37°C for 1 hour on a plate shaker. Subsequently, the wells were washed, TMB solution was added, and the reaction was terminated with 10% sulfuric acid solution after staining. The staining in the wells was recorded at 450 nm absorption using a Tecan plate fluorescence reader.

[0345] Prior to ELISA, samples taken from storage and reference samples were diluted 500-fold with buffer to a target concentration of 50 ng / ml, and the concentration of the test antibody was determined based on the ELISA results. Antibody stability was expressed as the ratio of the concentration of the sample taken from storage to the original sample concentration multiplied by 100%. A concentration reduction of no more than 60% in the incubated sample was considered sufficient for further studies (Table 6).

[0346] Table 6. Relative stability of antibodies stored in human serum at +37°C for 14 days.

[0347] This table shows the percentage of full-length antibody molecules remaining in serum after incubation, compared to the antibody content in the sample before incubation, and also shows the antibody concentration. It demonstrates that the anti-TRBV5-1 antibody according to the present invention is stable when stored in human serum.

[0348] Example 10. Testing the stability of the antibody.

[0349] HPLC was used to determine the differences in test antibody samples before and after heating at 50°C for 48 hours. Peak areas and their ratios for each product were evaluated in pairs. The comparison standard was the change in peak area and peak area ratio between the product stored at 4°C and the same product incubated at 50°C for 48 hours.

[0350] Table 7 shows the changes in peak area for the target molecule.

[0351] Table 7: Changes in the monomer ratio in antibody products subjected to heat stress.

[0352] Therefore, antibody 04-002 has higher thermal stability compared to antibody 01-010.

[0353] Example 11: The ability of antibodies to induce ADCC in reporter cell assays.

[0354] The assay used the Jurkat-NFAT-Luc-CD16 cell line, which stably expresses the firefly luciferase gene, which is expressed under the control of the NFAT promoter and contains CD16 on its surface. The J.RT3-T3.5-TRBV5-1 cell line, which stably expresses the T cell receptor β chain variable domain including the TRBV5-1 region, was used as the target cell.

[0355] The assays were performed in white 96-well plates designed for luminescence assays. Each well contained 25,000 Jurkat-NFAT-Luc-CD16 effector cells, 8,000 J.RT3-T3.5-TRBV5-1 target cells, and test antibody at concentrations indicated in the graph. The final volume of cell suspension and antibody in each well was 100 µl; all suspension components were prepared in RPMI-1640 medium containing 5% FBS and 2 mM glutamine. After adding all components, the plates were incubated at 37°C and 5% CO2 for 5 hours. The luminescence intensity in the wells was then measured using the One-Glo Luciferase Evaluation Kit (Promega). Measurements were performed on a plate reader.

[0356] The antibody-dependent cell-mediated cytotoxicity induced by antibodies 04-002, 04-004, 04-009, 04-013, 04-016, 04-024, 04-042, 04-045, 04-046, 04-047, 03-028, 03-015, 03-036, 03-038, 03-026, 03-008, 03-010, 03-023, 03-024, and 03-027 in the reporter molecule assay is shown in Table 8. The half-maximal effective antibody concentration (EC50) is shown in Table 8.

[0357] Table 8. Antibody-dependent cell-mediated cytotoxicity of the antibodies according to the present invention in reporter molecule assays.

[0358] Example 12. The ability of antibodies to induce ADCP in reporter cell assays.

[0359] The reporter cell assay used to test the antibody-induced ADCP capability employed the Jurkat-NFAT-Luc-CD64 cell line, which stably expresses surface CD64 and contains the firefly luciferase gene expressed under the control of the NFAT promoter; J.RT3-T3.5-TRBV5-1 cells were used as target cells, which stably express the T cell receptor β chain variable domain including the TRBV5-1 segment.

[0360] The assay was performed in white 96-well plates designed for luminescence assays. Each well contained 30,000 Jurkat-NFAT-Luc-CD64 effector cells, 30,000 J.RT3-T3.5-TRBV5-1 target cells, and the test antibody. Rituximab, an anti-CD20 antibody with ADCP activity, was used as a positive control. The final volume of cell suspension and antibody in each well was 100 µl, and all components of the suspension were prepared in RPMI-1640 medium containing 5% FBS. After adding all components, the plates were incubated at +37°C, 5% CO2 for 16 hours, and then the luminescence intensity in the wells was measured using the One-Glo Luciferase Assay Kit (Promega). Luminescence was measured using a plate reader.

[0361] The results show that antibodies 04-002 and 03-010 did not induce ADCP in the reporter cell assay. The mean and standard deviation for both replicates are shown. Figure 5 ).

[0362] Example 13. Analysis of nonspecific activation of NFAT signaling cascade by antibodies 03-010, 03-015, 04-002, 04-016 and 04-024.

[0363] The reporter cell assay for testing the activation properties of the antibody used the J.RT3-T3.5-TRBV5-1-NFAT-Luc cell line, which stably expresses a variable domain of the T cell receptor β chain containing the TRBV5-1 region and includes the firefly luciferase gene expressed under the control of the NFAT promoter. The assay was performed in white 96-well plates designed for luminescence assays. Each well contained 50,000 J.RT3-T3.5-TRBV5-1-NFAT-Luc cells with the test antibody. As a control, the anti-CD3 antibody OKT3 was used, pre-incubated with a cross-linked antibody targeting the Fc portion of the antibody. The final volume of cell suspension and antibody in the wells was 100 µl; all suspension components were prepared in RPMI-1640 medium containing 5% FBS and 2 mM glutamine. After adding all components, the plate was incubated at +37°C and 5% CO2 for 16 hours. Then, the luminescence intensity in the wells was measured using the One-Glo Luciferase Assay Kit (Promega). Luminescence was measured using a plate reader.

[0364] The results show that antibodies 03-010, 03-015, 04-002, 04-016, and 04-024 did not induce significant activation in the J.RT3-T3.5-TRBV5-1-NFAT-Luc reporter cell line. Mean values ​​with standard deviations are provided. Figure 6 and 7 ).

[0365] Example 14: Analysis of ADCC activity of antibodies 04-002, 04-016, 04-024 and 03-010 in suspension of peripheral blood mononuclear cells.

[0366] PBMC-based cell assays were performed to evaluate the ability of the antibodies to induce ADCC under real-world conditions. Assays were performed in 96-well plates for suspension culture. Each well contained 500,000 freshly isolated PBMCs from healthy donors, along with biotinylated antibodies at specified concentrations indicated in the graph. The final volume of cell suspension was 200 μl per well. All suspension components were prepared in RPMI-1640 medium containing 5% FBS and 2 mM glutamine. After mixing the PBMCs and antibodies, the plates were incubated at 37°C and 5% CO2 for 24 hours. Next, the proportion of TRBV5-1 positive T cells in the suspension was determined. For this purpose, cell pellets from each well were stained with the corresponding anti-TRBV5-1 biotinylated antibody at a concentration of 1 μg / ml, followed by staining with streptavidin labeled with PE fluorescence. The proportion of T cells in the suspension was determined by direct staining with a fluorescently labeled antibody against CD3. Analysis was performed using flow cytometry.

[0367] The antibody 04-002, 04-016, 04-024, and 03-010 were shown to induce antibody-dependent cell-mediated cytotoxicity in human PBMCs. The half-maximal effective antibody concentration (EC50) was shown at [data missing]. Figure 8 middle.

[0368] Example 15: Analysis of the depletion of native TRBV5-1 T cells in human PBMCs by real-time PCR.

[0369] To analyze the depletion of T cells expressing the variable domain of the T cell receptor β chain containing the TRBV5-1 region by real-time PCR (qPCR), one million cells were obtained from each sample incubated in the presence of candidate 04-002, and from each intact sample prepared as described in Example 14. The cell pellet was placed in RLT buffer (Qiagen), and RNA was extracted from it using the Qiagen RNeasy microkit reagent according to the manufacturer's protocol. The amount of isolated RNA was determined spectrophotometrically on a NanoDrop 8000 instrument (Thermo Scientific). The obtained RNA was qualitatively analyzed using denaturing RNA gel electrophoresis. Genomic DNA residues were removed using a DNase (New England Biolabs). cDNA synthesis using an RNA matrix was performed using the SuperScript IV First-Strand Synthesis System (Invitrogen) according to the manufacturer's protocol. 1 μg of RNA was used for each reverse transcription reaction. For quantitative PCR, the SYBR Green I Quantitative RT-PCR Kit (Syntol) was used. PCR was performed three times per sample using the same technique. 3-Glyceraldehyde phosphate dehydrogenase ( GAPDH Genes were used as references to standardize the obtained quantitative evaluation values. Real-time PCR was performed in a StepOne™ Real-Time PCR System (Applied Bioscience). TRBV5-1 expression levels were determined using Pfaffl, with corrections for amplification efficiency.

[0370] Therefore, it was shown that ( Figure 9 Antibody 04-002 induces antibody-dependent cell-mediated cytotoxicity, clearing T cell populations carrying the variable domain of the T cell receptor β chain containing the TRBV5-1 segment on their surface.

[0371] Example 16: The ability of antibodies to induce CDC in reporter cell assays.

[0372] The CDC assay used the J.RT3-T3.5-TRBV5-1 cell line. The assay was performed in 96-well plates. Each well contained a suspension of 50,000 J.RT3-T3.5-TRBV5-1 cells, along with a test antibody at a specified concentration and human serum complement diluted 1:4. The final volume of the cell suspension in each well was 150 µl, and all components of the suspension were prepared in RPMI-1640 medium containing 0.1% BSA. The plates were then incubated at 37 °C with 5% CO2 for 4 hours. 15 µl of AlamarBlue reagent was added to each well, and the plates were incubated at +37 °C with 5% CO2 for 16 hours.

[0373] Fluorescence was measured at an excitation wavelength of 544 nm and an emission wavelength of 590 nm using a plate reader.

[0374] The results showed that antibodies 04-002 and 03-010 did not induce complement-dependent cytotoxicity in the assay. Figure 10 The graph shows the mean along with the standard deviation.

[0375] Example 17: Analysis of autocytotoxic activity mediated by anti-TRBV5-1 antibody.

[0376] Autologous cytotoxicity assays were performed in 96-well culture plates used for suspension culture. Suspensions contained 300,000 freshly isolated PBMCs from healthy donors per well, along with antibodies at specified concentrations as shown in the graph, with a final volume of 150 µl of cell suspension in each well. All suspension components were prepared in RPMI-1640 medium containing 5% FBS and 2 mM glutamine. After mixing the PBMCs and antibodies, the plates were incubated at 37°C and 5% CO2 for 16 hours. The proportions of CD56+, CD19+, CD3+, and CD14+ subsets of PBMCs in the suspension were then measured by direct staining of the suspension with fluorescently labeled antibodies against the corresponding CDs and subsequent cell analysis using flow cytometry. Anti-CD20 antibody (GA101), which induces B cell exhaustion at concentrations ranging from 1 to 100 μg / ml, was used as a positive control.

[0377] The graph shows the mean and standard deviation for two replicates. Figure 11-14 ).

[0378] Conclusion: Antibody 04-002 did not exhibit the ability to deplete NK cell or B cell populations, and it did not affect the total T cell population except for those T cells carrying TRBV5-1. The results indicate that... Figure 13According to the T cell levels, the antibody 04-002 concentration decreased by about 5% at concentrations ranging from 1 to 100 μg / ml.

[0379] Example 18: Analysis of the properties of antibody effectors in blood and spleen samples in an in vivo NSIG mouse model.

[0380] Two groups of NSIG mice were administered human peripheral blood mononuclear cells from healthy donors (administered into the retroorbital sinus, 10 7 (10 cells / mouse; administration volume: 200 µl). On day 2, day 7, and day 16, a group of mice was administered anti-TRBV5-1 antibody 04-002 (n=9) (intraperitoneal administration, dose: 10 μg / kg; administration volume: 200 µl). No antibody was administered to the control group (n=8). On day 18 post-administration, the percentage of TRBV5-1 positive cells in blood and spleen samples was evaluated by flow cytometry. TRBV5-1 positive cells were detected in human CD45 and CD3 positive cell populations. Figure 15 (Blood sample) and 16 (spleen sample) show the percentage of TRBV5-1 positive cells per animal, median ± interquartile range. Blood and spleen samples from NSIG mice with circulating human peripheral blood mononuclear cells are shown after administration of monoclonal antibody 04-002, exhibiting significant depletion of a CD3-positive cell population expressing the variable domain of the T-cell receptor β chain containing the TRBV5-1 segment.

Claims

1. A monoclonal antibody or antigen-binding fragment thereof that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain, comprising: (a) Contains the following light chain variable structural domains: (i) Having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) CDR2 having the amino acid sequence EIX1KLX2S, wherein X1 = T or S; X2 = A, F, or M; as well as (iii) CDR3 having the amino acid sequence QQWNYPХ3LX4, in which X3 = L, R, or Y; X4 = S or T; as well as (b) Contains the following heavy-chain variable structural domains: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) CDR2 having the amino acid sequence YINPX5X6GRTGYNQKX7QX8, in which X5 = Y or W; X6 = Q or N; X7 = F or L; X8 = A or G; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO:

18.

2. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises a CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO:

7.

3. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises a CDR3 having an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO:

11.

4. The monoclonal antibody or its antigen-binding fragment according to claim 1, wherein the light chain variable domain comprises: (i) Having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7; as well as (iii) Having a CDR3 with an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO:

11.

5. The monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1-4, comprising: (i) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (ii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (iii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (iv) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (v) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; or (vi) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; or (vii) contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 10; or (viii) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; or (ix) contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 10; or (x) contains the following light chain variable structure domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO:

9.

6. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO:

17.

7. The monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 or 6, wherein the heavy chain variable domain comprises: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO:

18.

8. The monoclonal antibody or its antigen-binding fragment according to claim 7, comprising: (i) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ii) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iii) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iv) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (v) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO:

18.

9. The monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1-8, comprising: (a) Contains the following light chain variable structural domains: (i) Having a CDR1 with an amino acid sequence selected from the following: SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7; as well as (iii) A CDR3 having an amino acid sequence selected from the following: SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: (i) CDR1 having the amino acid sequence of SEQ ID NO: 12; (ii) A CDR2 having an amino acid sequence selected from the following: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17; as well as (iii) CDR3 having the amino acid sequence of SEQ ID NO:

18.

10. The monoclonal antibody or its antigen-binding fragment according to claim 9, comprising: (i) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (iv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (v) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 14, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (vi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 13, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (vii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (viii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ix) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (x) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 15, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xiii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 10; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xiv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 5, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xv) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xvi) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xvii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 11; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xviii) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 8; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 16, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (ixx) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 6, and CDR3 having the amino acid sequence of SEQ ID NO: 10; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO: 18; or (xx) (a) Contains the following light chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 9; and (b) Contains the following heavy-chain variable structural domains: CDR1, having the amino acid sequence of SEQ ID NO: 12, CDR2 having the amino acid sequence of SEQ ID NO: 17, and CDR3 having the amino acid sequence of SEQ ID NO:

18.

11. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the light chain variable domain comprises an amino acid sequence selected from the following: SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27 or SEQ ID NO:

28.

12. The monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises an amino acid sequence selected from the following: SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO:

34.

13. The monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 or 11-12, wherein: (a) The light chain variable domain comprises an amino acid sequence selected from the following: SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27 or SEQ ID NO: 28; and (b) The heavy chain variable domain comprises an amino acid sequence selected from the following: SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 or SEQ ID NO:

34.

14. The monoclonal antibody or its antigen-binding fragment according to claim 13, wherein: (i) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 19, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 29; or (ii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 20, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 29; or (iii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 21, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 30; or (iv) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 22, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 30; or (v) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 23, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 30; or (vi) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 23, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 31; or (vii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 19, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 32; or (viii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 22, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 32; or (ix) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 24, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 32; or (x) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 26, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 32; or (xi) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 21, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 19, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xiii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 25, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 34; or (xiv) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 20, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xv) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 22, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xvi) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 24, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xvii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 26, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (xviii) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 23, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 33; or (ixx) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 27, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO: 34; or (xx) (a) The light chain variable domain contains the amino acid sequence of SEQ ID NO: 28, and (b) The heavy chain variable domain contains the amino acid sequence of SEQ ID NO:

34.

15. The monoclonal antibody according to any one of claims 1-14, wherein the antibody that specifically binds to the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is a full-length IgG antibody.

16. The monoclonal antibody of claim 15, wherein the full-length IgG antibody has a human IgG1, IgG2, IgG3 or IgG4 isotype.

17. The monoclonal antibody of claim 16, wherein the full-length IgG antibody has a human IgG1 isotype.

18. The monoclonal antibody of claim 17, wherein the antibody comprises a deletion of 446G and 447K in the CH3 region according to the EU numbering scheme of antibody amino acids.

19. The monoclonal antibody of claim 17, wherein the antibody comprises S239D and I332E mutations according to the EU numbering scheme for antibody amino acids.

20. The monoclonal antibody of claim 17, wherein the antibody comprises G236A, S239D, and I332E mutations according to the EU numbering scheme for antibody amino acids.

21. The monoclonal antibody of claim 1, comprising a light chain containing an amino acid sequence selected from the following: SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 or SEQ ID NO:

44.

22. The monoclonal antibody of claim 1, comprising a heavy chain containing an amino acid sequence selected from the following: SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 or SEQ ID NO:

52.

23. The monoclonal antibody according to any one of claims 1 or 21-22, comprising: (i) (a) A light chain comprising an amino acid sequence selected from: SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43 or SEQ ID NO: 44, and (b) A heavy chain comprising an amino acid sequence selected from the following: SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51 or SEQ ID NO:

52.

24. The monoclonal antibody according to claim 23, comprising: (i) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 45; or (ii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 36, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 46; or (iii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 37, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47; or (iv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 47; or (v) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 48; or (vi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 49; or (vii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (viii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (ix) (a) A light chain containing the amino acid sequence of SEQ ID NO: 40, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (x) (a) A light chain containing the amino acid sequence of SEQ ID NO: 42, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 50; or (xi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 37, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 35, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xiii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 41, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52; or (xiv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 36, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xv) (a) A light chain containing the amino acid sequence of SEQ ID NO: 38, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xvi) (a) A light chain containing the amino acid sequence of SEQ ID NO: 40, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xvii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 42, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (xviii) (a) A light chain containing the amino acid sequence of SEQ ID NO: 39, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 51; or (ixx) (a) A light chain containing the amino acid sequence of SEQ ID NO: 43, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 52; or (xx) (a) A light chain containing the amino acid sequence of SEQ ID NO: 44, and (b) A heavy chain containing the amino acid sequence of SEQ ID NO:

52.

25. A nucleic acid encoding an antibody or an antigen-binding fragment thereof according to any one of claims 1-24.

26. The nucleic acid according to claim 25, wherein the nucleic acid is DNA.

27. An expression vector comprising the nucleic acid according to any one of claims 25-26.

28. A method for generating host cells to produce an antibody or an antigen-binding fragment thereof according to any one of claims 1-24, comprising cell transformation via the vector according to claim 27.

29. A host cell for producing an antibody or an antigen-binding fragment thereof according to any one of claims 1-24, comprising a nucleic acid according to any one of claims 25-26.

30. A method for producing an antibody or an antigen-binding fragment thereof according to any one of claims 1-24, comprising culturing a host cell according to claim 29 in a growth medium under conditions sufficient for producing the antibody, followed by isolation and purification of the resulting antibody.

31. A pharmaceutical composition comprising, in combination with one or more pharmaceutically acceptable excipients, a therapeutically effective amount of an antibody or an antigen-binding fragment thereof according to any one of claims 1-24.

32. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to any one of claims 1-24, and at least one other therapeutically active compound.

33. The pharmaceutical composition of claim 32, wherein the other therapeutically active compound is an antibody, an insulin preparation, a small molecule, a hormone therapy agent, or any combination thereof.

34. The pharmaceutical composition according to any one of claims 31 or 32, intended for the treatment or prevention of diseases or conditions mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain.

35. The pharmaceutical composition of claim 34, wherein the disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain is selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

36. A method for treating or preventing a disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain, comprising administering to a subject in need of such treatment or prevention a therapeutically effective amount of an antibody or antigen-binding fragment thereof according to any one of claims 1-24, or a pharmaceutical composition according to any one of claims 31 or 32.

37. The method of claim 36 for treating or preventing a disease or condition, wherein the disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T-cell receptor β chain is selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

38. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1-24, or the pharmaceutical composition according to any one of claims 31-32, for the treatment or prevention of a disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain in a subject requiring such treatment or prevention.

39. The use according to claim 38, wherein the disease or condition mediated by the TRBV5-1 segment of the variable domain of the human T cell receptor β chain is selected from the following: type 1 diabetes, delayed progression of type 1 diabetes from stage 2 to stage 3, hyperglycemia, impaired glucose tolerance, prediabetes, rejection of nickel implants or prostheses, sarcoidosis, beryllium disease, celiac disease, or herpetic dermatitis.

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