Anti-s100a8 / a9 antibody and use thereof

Abstract

An anti-S100A8 / A9 antibody and a use thereof. The antibody or an antigen-binding fragment and mutant thereof can block the biological function of S100A8 / A9. The antibody or the antigen binding fragment has prospects of being developed into a clinical drug for reducing the myocardial infarction area of AMI patients and reducing the occurrence of heart failure, thereby benefiting the AMI patients for life. The antibody or the antigen binding fragment has important clinical significance and development value.

Description

Anti-S100A8 / A9 antibodies and uses thereof

[0001] Cross-references

[0002] This application refers to Chinese Patent Application No. 202410175455.0, “Antibodies binding to S100A8 and S100A9 heterodimers or tetramers and their uses,” filed on February 7, 2024, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present application relates to the field of molecular biology, and specifically to a fully human antibody or an antigen-binding fragment thereof and uses thereof. Background Art

[0004] Cardiovascular disease (CVD) is the leading cause of death and the largest global burden of disease. In China, 2 out of every 5 deaths are attributable to CVD, with over half of these deaths related to coronary heart disease (CHD). Acute myocardial infarction (AMI), a severe form of CHD, is a leading cause of death and disability. Acute and persistent ischemia and hypoxia in the coronary arteries cause myocardial cell necrosis. Current treatments prioritize swift recanalization of the infarct-related artery to restore myocardial blood perfusion. Thrombolysis, percutaneous coronary intervention (PCI), or thrombolysis followed by PCI are options tailored to the patient's condition. The popularity of PCI surgery has improved the treatment of AMI, and reperfusion effectively prevents further necrosis of myocardial cells. However, it still faces two problems: First, the average time for AMI patients to arrive at the hospital after onset is 5.5 hours, so the blocked blood vessels are usually recanalized 6 hours after the onset of myocardial infarction. During this period, myocardial cells have already undergone necrosis due to ischemia and hypoxia, but there are no early treatment drugs that can reduce the area of ​​myocardial infarction; second, reperfusion injury (I / R) will also be induced after the blood vessels are recanalized. In addition, part of the myocardium has already undergone necrosis before recanalization. Therefore, the occurrence of decreased cardiac function and even heart failure remains a prominent problem, and the current clinical drug application cannot improve this situation.

[0005] Cardiac damage caused by AMI is the result of the synergistic effects of multiple cellular and pathological processes. For example, both AMI and I / R induce the infiltration of a large number of inflammatory cells. In animal studies, inhibiting inflammation can reduce post-MI damage and improve cardiac function. However, multiple clinical trials of anti-inflammatory treatments for patients with MI have failed, suggesting that broad-spectrum inflammation suppression is undesirable in the treatment of ischemic heart disease and that more specific and targeted targets are needed.

[0006] S100A8 and S100A9 (also known as MRP8 and MRP14) belong to the S100 family of calcium-binding proteins. Human S100A8 consists of 98 amino acids with a molecular weight of 10.8 kDa; S100A9 consists of 113 amino acids with a molecular weight of 13.2 kDa. Both proteins can form heterodimers, heterotetramers, and oligomers, predominantly in the dimer form, but each can also form homomers. Due to the poor stability of homodimers, they typically function as heterodimers and tetramers in pathophysiological conditions. S100A8 / S100A9 are primarily expressed by neutrophils, comprising approximately 40% of their cytoplasmic proteins. When inflammatory cells are stressed, S100A8 / S100A9 are released as alarmins, acting on Toll-like receptor 4 (TLR4) and the receptor for advanced glycation end products (RAGE) to regulate immune inflammatory responses.

[0007] Our previous work (Circulation, 2019, 140(9):751-764) used transcriptome sequencing to find that the most obvious changes in early myocardial ischemia in mice were in inflammatory response-related genes, and among the inflammatory-related differentially expressed genes, S100A8 and S100A9 were the most significantly elevated, reaching a peak 6 hours after I / R, then gradually declining, and basically returning to baseline levels after 7 days. Subsequent studies by multiple teams (Circulation, 2020, 141(13):1080-1094; Eur Heart J, 2019, 40(32):2713-2723) have all verified our results: S100A8 and S100A9 are released in large quantities in the early stages of myocardial infarction. At the same time, we also observed that serum S100A8 / A9 levels in AMI patients were significantly elevated one day after PCI, and elevated S100A8 / A9 levels were associated with the incidence of major adverse cardiovascular events (MACE), making it an independent predictor of long-term MACE in AMI patients (Circulation, 2019, 140(9):751-764). In addition, we also demonstrated that S100A8 / A9 acts on TLR4 in cardiomyocytes, regulating mitochondrial dysfunction and thus mediating myocardial injury (Circulation, 2019, 140(9):751-764). S100A9 gene knockout, S100A9 neutralizing antibodies or inhibitors can reduce cardiac injury and adverse remodeling after myocardial infarction or myocardial ischemia / reperfusion in mice, and improve cardiac function (Circulation, 2019, 140(9):751-764; Circulation, 2020, 141(13):1080-1094; Eur Heart J, 2019, 40(32):2713-2723). The above results confirm that S100A8 / A9 is an excellent target for early intervention treatment of myocardial infarction and myocardial ischemia / reperfusion injury, and is more specific and effective than broad-spectrum anti-inflammatory treatment.

[0008] However, the clinical application of S100A8 / A9 as a target still faces numerous challenges. First, the homology between human and mouse S100A8 / A9 is low, making previous studies of neutralizing antibodies against mice difficult to translate to humans. Furthermore, commercially available S100A9 antibodies lack fully human or humanized monoclonal antibodies. Second, S100A8 / A9 primarily exerts its cardiac damage-inducing effects as heterodimers and heterotetramers, and previous neutralizing antibodies targeting S100A9 were not specific for these dimers or tetramers. Therefore, the development of neutralizing antibodies against human S100A8 / A9 and their in vivo validation of their therapeutic efficacy are crucial.

[0009] It should be noted that the approaches described in this section are not necessarily approaches that have been previously conceived or employed. Unless otherwise indicated, it should not be assumed that any approach described in this section is prior art simply because it is included in this section. Similarly, unless otherwise indicated, the issues mentioned in this section should not be considered to have been recognized in any prior art.

[0010] Summary of the Invention

[0011] In order to solve the above technical problems, the present application provides a fully human antibody or an antigen-binding fragment thereof, wherein the complementarity determining region (CDR) defined by the KABAT system has the following amino acid sequence: the amino acid sequence of the heavy chain CDR1 includes GX1TX2SX3X4X5X6X7, wherein X1 is F or Y, X2 is F, H, Y or W, X3 is S or D, X4 is Y, H or F, X5 is A or V, X6 is M, I or L, and X7 is S or A; the amino acid sequence of the heavy chain CDR2 includes AISGX8GGSX9X 10 YX 11 X 12 SVKG, where X8 is S or H, X9 is T or H, X 10 is Y, W or F, X 11 A, H, V or L, X 12 is D, A or H; the amino acid sequence of the heavy chain CDR3 includes KX 13 RPX 14 RX 15 X 16 DX 17 , X 13 is Y, F or H, X 14 S, T or H, X 15 V or A, X 16 F, Y, H or W, X 17 is S, A, F, C, H or Y; the amino acid sequence of the light chain CDR1 includes SGDALX 18 DKX 19 X 20 X 21 , where X 18 G or H, X 19 Y or F, X 20 A or M, X 21 is S or A; the amino acid sequence of the light chain CDR2 includes X 22 X 23 X 24 X 25 RX 26 X 27 , where X 22 E or Q, X 23 D, H or Q, X 24is S, A or H, X 25 K, H, R, P or N, X 26 is P, Q, D or N, X 27 is S, D, Y or W; and the amino acid sequence of the light chain CDR3 includes X 28 SNDX 29 DX 30 X 31 W, where X 28 For Q or D, X 29 A, V, I or L, X 30 S or A, X 31 V, Y, W or R.

[0012] According to one embodiment of the present application, a polynucleotide is also provided, which encodes the antibody or antigen-binding fragment thereof described in the present application.

[0013] According to one embodiment of the present application, a recombinant vector is also provided, which comprises the polynucleotide described in the present application.

[0014] According to one embodiment of the present application, a host cell is also provided, wherein the host cell comprises the polynucleotide described in the present application and / or the recombinant vector described in the present application.

[0015] According to one embodiment of the present application, a pharmaceutical composition is also provided, comprising the antibody or antigen-binding fragment thereof described in the present application.

[0016] According to one embodiment of the present application, a kit is also provided, which includes the antibody or antigen-binding fragment thereof described in the present application.

[0017] According to one embodiment of the present application, a multispecific antibody is also provided, which comprises the antibody or antigen-binding fragment thereof described in the present application, and further comprises one or more second antibodies or antigen-binding fragments thereof that specifically bind to other antigens.

[0018] According to one embodiment of the present application, an antibody conjugate is also provided, which comprises the antibody or antigen-binding fragment described in the present application and a second functional structure, wherein the second functional structure is selected from Fc, a radioactive isotope, a structural portion that extends half-life, a detectable marker and a drug.

[0019] According to one embodiment of the present application, there is also provided use of the antibody or antigen-binding fragment thereof, the multispecific antibody, and / or the antibody conjugate described herein in the preparation of drugs, reagents, and kits for detecting, alleviating, preventing, or treating diseases.

[0020] According to one embodiment of the present application, there is also provided the use of the antibody or antigen-binding fragment thereof, the multispecific antibody, and / or the antibody conjugate described herein in the preparation of drugs, reagents, and kits for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function.

[0021] According to one embodiment of the present application, a method for detecting, alleviating, preventing or treating a disease is also provided, comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof described herein, the pharmaceutical composition described herein, the kit described herein, the multispecific antibody described herein, and / or the antibody conjugate described herein.

[0022] According to one embodiment of the present application, a method for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function is also provided, the method comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof described herein, the pharmaceutical composition described herein, the kit described herein, the multispecific antibody described herein, and / or the antibody conjugate described herein.

[0023] The present application provides a fully human monoclonal antibody that can block the biological function of the S100A8 / A9 heterodimer. The present application utilizes established humanized mouse models of S100A8 and S100A9 to verify the in vivo therapeutic effect of the antibody on ischemic heart disease. For example, application of the antibody in the acute phases of MI and I / R can inhibit cardiac damage caused by the target molecule, reduce the area of ​​myocardial infarction and the degree of late fibrosis, and improve cardiac function in experimental animals. Therefore, the antibody or antigen-binding fragment provided in the present application is expected to be developed into a clinical drug to reduce the area of ​​myocardial infarction in AMI patients, reduce the incidence of heart failure, and provide lifelong benefits to AMI patients, thus having important clinical significance and development value.

[0024] It should be understood that the content described in this section is not intended to identify the key or important features of the embodiments of the present application, nor is it intended to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings illustrate exemplary embodiments and constitute a part of the specification. Together with the description of the specification, they serve to explain exemplary implementation of the embodiments. The illustrated embodiments are for illustrative purposes only and do not limit the scope of the claims. Throughout the drawings, the same reference numerals designate similar, but not necessarily identical, elements.

[0026] FIG1 is a graph showing the binding activity of the four candidate monoclonal antibodies in Example 2 to human S100A8 / A9 (hS100A8 / A9) antigen.

[0027] FIG2 is a graph showing the binding specificity of the anti-S100A8 / A9 monoclonal antibody to hS100A8 / A9 homologous proteins in Example 2.

[0028] FIG3 is a graph showing the binding specificity of the anti-S100A8 / A9 monoclonal antibody to unrelated proteins in Example 2.

[0029] FIG4 is a graph showing the results of species-specific detection of anti-S100A8 / A9 monoclonal antibodies in Example 2.

[0030] FIG5A is a graph showing the stability of the anti-S100A8 / A9 monoclonal antibody in PBS in Example 2; FIG5B is a graph showing the stability of the anti-S100A8 / A9 monoclonal antibody in C57BL / 6 mouse serum (m-Serum) in Example 2.

[0031] FIG6 is a diagram showing the results of epitope analysis of the binding of anti-S100A8 / A9 monoclonal antibodies to hS100A8 / A9 in Example 3.

[0032] Figure 7 shows the results of the inhibitory effect of anti-S100A8 / A9 mAbs on the release of inflammatory factors from human PBMCs stimulated by hS100A8 / A9 in Example 4. Figure 7A shows the results of the inhibitory effect of anti-S100A8 / A9 mAbs on the release of IL-6 from human PBMCs stimulated by hS100A8 / A9 in Example 4. Figure 7B shows the results of the inhibitory effect of anti-S100A8 / A9 mAbs on the release of TNFα from human PBMCs stimulated by hS100A8 / A9 in Example 4.

[0033] FIG8 is a graph showing the proliferative toxicity of the anti-S100A8 / A9 monoclonal antibody in Example 4 on mouse lymphoma cells EL-4.

[0034] FIG9 shows the results of Example 5 showing that the anti-S100A8 / A9 monoclonal antibody can block the binding of hS100A8 / A9 to TLR4 ( FIG9A ) or RAGE ( FIG9B ) on the surface of the engineered cell line.

[0035] FIG10 shows the results of the anti-S100A8 / A9 monoclonal antibody blocking the binding of hS100A8 / A9 to PBMC cells ( FIG10A ) and THP-1 cells ( FIG10B ) in Example 5. FIG10B shows the results of the anti-S100A8 / A9 monoclonal antibody blocking the binding of hS100A8 / A9 to PBMC cells ( FIG10A ) and THP-1 cells ( FIG10B ).

[0036] FIG11 is a graph showing the in vitro biological activity of the genetically engineered single-point mutants of the anti-S100A8 / A9 monoclonal antibody in Example 6.2.

[0037] FIG. 12 ( FIG. 12A and FIG. 12B ) shows the results of the in vitro biological activities of the genetically engineered combination mutants of the anti-S100A8 / A9 monoclonal antibodies in Example 6.4.

[0038] FIG13 is a graph showing the metabolic status of the anti-S100A8 / A9 monoclonal antibody in mice in Example 7.

[0039] FIG14 is a graph showing the effects of low, medium and high doses of anti-S100A8 / A9 monoclonal antibodies on left ventricular ejection fraction in S100A8 / A9 humanized mice after myocardial infarction in Example 8.

[0040] FIG15 is a graph showing the effects of low, medium and high doses of anti-S100A8 / A9 monoclonal antibodies on the infarct size of S100A8 / A9 humanized mice after myocardial infarction in Example 8.

[0041] FIG16 is a graph showing the effect of the anti-S100A8 / A9 monoclonal antibody administered 2, 4, 8, and 10 hours after surgery on the left ventricular ejection fraction of S100A8 / A9 humanized mice after myocardial infarction in Example 8.

[0042] FIG17 is a graph showing the effect of the anti-S100A8 / A9 monoclonal antibody administered 2, 4, 8, and 10 hours after surgery on the infarct size of S100A8 / A9 humanized mice after myocardial infarction in Example 8.

[0043] FIG18 is a graph showing the effect of the anti-S100A8 / A9 monoclonal antibody on the left ventricular ejection fraction of S100A8 / A9 humanized mice after myocardial ischemia / reperfusion in Example 8. DETAILED DESCRIPTION

[0044] Unless otherwise indicated, all numbers used in this specification and claims to represent content, concentration, ratio, mass, volume, time, temperature, thickness, technical effect, etc. should be understood as being modified by the term "about" or "approximately" in any case. Therefore, unless otherwise indicated, the numerical parameters listed in the following specification and the appended claims are approximate values. For those skilled in the art, it can vary according to the desired properties and effects sought to be obtained through this disclosure, and each numerical parameter should be interpreted according to the number of significant digits and conventional rounding methods or in a manner understood by those skilled in the art.

[0045] Although the numerical ranges and parameters setting forth the broad scope of the present disclosure are approximations, the numerical values ​​set forth in the specific examples are provided as precisely as possible. However, any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in its respective testing measurements. Every numerical range given herein will include every narrower numerical range that falls within that broader numerical range, as if each narrower numerical range were expressly written herein.

[0046] Unless otherwise specified or contradicted by the context, the terms or expressions used in this article should be read in conjunction with the entire content of this article and as understood by one of ordinary skill in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0047] As used herein, the expression "A and / or B" includes three cases: (1) A; (2) B; and (3) A and B. The expression "A, B, and / or C" includes seven cases: (1) A; (2) B; (3) C; (4) A and B; (5) A and C; (6) B and C; and (7) A, B, and C. The meanings of similar expressions can be deduced analogously.

[0048] As used herein, "nucleic acid" and "polynucleotide" are used interchangeably to refer to polymeric forms of nucleotides of any length, including deoxyribonucleotides, ribonucleotides, combinations thereof, and analogs thereof.

[0049] As used herein, "polypeptide" and "peptide" are used interchangeably to refer to amino acid polymers of any length. Thus, polypeptides, oligopeptides, proteins, antibodies, and enzymes are all included within the definition of polypeptide.

[0050] As used herein, a "fragment" of a sequence refers to a portion of the sequence. For example, a fragment of a nucleic acid sequence refers to a portion of the nucleic acid sequence, and a fragment of an amino acid sequence refers to a portion of the amino acid sequence.

[0051] As described in this application, sequence "identity" refers to the extent to which two (nucleotide or amino acid) sequences have the same residue at the same position in an alignment, and is usually expressed as a percentage. Preferably, identity is determined over the entire length of the sequences being compared. Thus, two copies of exactly the same sequence have 100% identity. Those skilled in the art will appreciate that sequence identity can be determined using algorithms such as Blast (Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402), Blast2 (Altschul et al. (1990) J. Mol. Biol. 215: 403-410), Smith-Waterman (Smith et al. (1981) J. Mol. Biol. 147: 195-197), and ClustalW.

[0052] As used herein, “S100A8 / S100A9,” “S100A8 / A9,” “S100A8 / S100A9 dimer,” “S100A8 / S100A9 heterodimer,” “S100A8 / S100A9 polypeptide,” “S100A8 / S100A9 protein,” “dimer of S100A8 and S100A9,” or “heterodimer of S100A8 and S100A9” and similar expressions have the same meaning and are used interchangeably to refer to a protein complex in the form of a heterodimer formed by calcium-binding protein A8 (S100A8) and calcium-binding protein A9 (S100A9) in the presence of calcium and zinc ions.

[0053] "Antibody" as described herein refers to a specific immunoglobulin for an antigenic site. Antibodies can be manufactured according to methods known in the art. The form of the antibody includes polyclonal or monoclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2 and Fv fragments), single-chain Fv (scFv) antibodies, multispecific antibodies (e.g., bispecific antibodies), monospecific antibodies, monovalent antibodies, chimeric antibodies, fully human antibodies, human antibodies, fusion proteins comprising the antigen-binding site of an antibody, and any other modified immunoglobulin molecules comprising an antigen-binding site, as long as the antibody exhibits the desired biological binding activity.

[0054] As used herein, "multispecific" refers to an antigen-binding protein that has multiple epitope specificities (i.e., it can specifically bind to two, three, or more different epitopes on a single biomolecule or can specifically bind to epitopes on two, three, or more different biomolecules). As used herein, "bispecific" refers to an antigen-binding protein that has two different antigen-binding specificities.

[0055] The precise amino acid sequence boundaries of a given complementarity determining region (CDR) or framework region (FR) can be readily determined using a number of numbering schemes well known in the art, including: Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th ed. Public Health Service, National Institutes of Health, Bethesda, Maryland ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding sitetopography," J. Mol. Biol. 262, 732-745 ("Contact" numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Dev Comp Immunol, 2003 Jan;27(1):55-77 (“IMGT” numbering scheme); Honegger A and Plückthun A, “Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool,” J Mol Biol, 2001 Jun 8;309(3):657-70 (“Aho” numbering scheme); and Martin et al., “Modeling antibody hypervariable loops: a combined algorithm,” PNAS, 1989, 86(23):9268-9272 (“AbM” numbering scheme).

[0056] In some embodiments, the boundary of CDR or FR is different depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, and the Chothia scheme is based on structural information. The numbering of Kabat and Chothia schemes is all based on the most common antibody region sequence length, wherein insertion (for example " 30a ") is provided by inserting letters and disappearance occurs in some antibodies. These two schemes are placed in different positions with certain insertions and deletions (indel), thereby producing different numbering. The Contact scheme is based on the analysis of complex crystal structure, and is similar to the Chothia numbering scheme in many aspects. The AbM scheme is a compromise between Kabat and Chothia definitions, and it is based on the scheme used by the AbM antibody modeling software of Oxford Molecular.

[0057] Therefore, unless otherwise specified, it should be understood that the "CDR" of a given antibody or its region (such as its variable region) covers the CDRs defined by any of the above schemes or other known schemes. For example, when specifying that a specific CDR (such as CDR3) contains a given amino acid sequence, it should be understood that such CDR can also have the sequence of the corresponding CDR (such as CDR3) defined by any of the above schemes or other known schemes. Similarly, unless otherwise specified, it should be understood that the FR of a given antibody or its region (such as its variable region) covers the FR defined by any of the above schemes or other known schemes. Unless otherwise specified, the numbering scheme used to define the boundaries of CDR and FR in this article adopts the Kabat scheme.

[0058] "Humanized" antibodies described in this application refer to antibodies in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A "humanized form" of a non-human antibody refers to a variant of the non-human antibody that has undergone humanization to generally reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. In some embodiments, some FR residues in the humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., an antibody derived from CDR residues), for example to restore or improve antibody specificity or affinity. Humanized antibodies and methods for preparing them are well known to those skilled in the art, see, for example, Almagro and Fransson, Front.Biosci.13:1619-1633 (2008). Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best fit" method; framework regions derived from the consensus sequence of human antibodies of a specific subgroup of light or heavy chain variable regions; human mature (somatic mutation) framework regions or human germline framework regions; and framework regions obtained by screening FR libraries.

[0059] As used herein, "gene" refers to the entire nucleotide sequence required to produce a polypeptide chain or functional RNA. Therefore, gene expression includes transcription and the stable accumulation of coding RNA (mRNA) or functional RNA derived from the gene, and also includes translation of mRNA into polypeptide or protein.

[0060] It should be noted that in the context of this application, upstream refers to the 5' end of the gene or the N-terminus of the protein, and downstream refers to the 3' end of the gene or the C-terminus of the protein, and from upstream to downstream is from 5' end to 3' end or N-terminus to C-terminus.

[0061] As used herein, "exogenous" or "heterologous" are used interchangeably to refer to an organism of a different origin than the native (original) organism, such as an organism from another species. As used herein, a "heterologous gene" or "exogenous gene" refers to a gene that does not naturally occur in the host organism and that is introduced into the host organism by gene transfer.

[0062] As used herein, a "vector" refers to a self-replicating DNA molecule that transfers an exogenous target gene into a host organism, and is often in the form of a circular double-stranded DNA molecule. Typical vectors include plasmids, viruses, bacteriophages, cosmids, and minichromosomes. Plasmids are the most common form of vector and are circular double-stranded DNA molecules that can accept exogenous nucleic acid fragments and replicate in prokaryotic or eukaryotic cells.

[0063] As used herein, the terms "expression vector" and "recombinant vector" are used interchangeably to refer to a vector containing an exogenous gene and regulatory elements for expression in a designated host organism. Introduction of an expression vector into an appropriate host organism enables the expression of the inserted gene of interest.

[0064] "Transformation" as used herein refers to the transfer of exogenous genes into a host organism, such as a host cell, resulting in genetically stable inheritance. The transformed gene can be in the form of a plasmid retained in the host organism, or it can be integrated into the host organism's genome. Host organisms containing transforming genes are referred to as "transgenic," "recombinant," "transformed," or "engineered" organisms. Expression vectors can be used to transform host organisms using conventional techniques well known to those skilled in the art. When the host is a prokaryotic organism, competent cells capable of absorbing DNA can be harvested after the exponential growth phase and treated with CaCl2, a step well known in the art. If desired, methods such as microinjection, electroporation, or liposome packaging can also be used. This is a well-known technique in the art and will not be described in detail herein.

[0065] As used herein, "relieve" and "treat" and their synonyms refer to the improvement of a disease, disorder, and / or condition. "Relieve" and "treat" can be an improvement in at least one measurable physical parameter, which is not necessarily recognizable to the patient. "Relieve" and "treat" can also be the inhibition of the progression of a disease, disorder, and / or condition physically (e.g., stabilizing recognizable symptoms), physiologically (e.g., stabilizing physical parameters), or both. "Relieve" and "treat" can also be the slowing down of the progression of a disease, disorder, and / or condition or its reversal.

[0066] As used herein, "prevent," "prevent," and its synonyms refer to delaying the onset of a particular disease, disorder, and / or condition, or symptoms associated with such disease, disorder, and / or condition, or reducing the risk of acquiring such disease, disorder, and / or condition.

[0067] In order to make the above-mentioned objects, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below.

[0068] Antibodies or antigen-binding fragments

[0069] According to one embodiment of the present application, a fully human antibody or an antigen-binding fragment thereof is provided, characterized in that its complementarity determining region (CDR) defined according to the KABAT system has the following amino acid sequence: the amino acid sequence of the heavy chain CDR1 includes GX1TX2SX3X4X5X6X7, wherein X1 is F or Y, X2 is F, H, Y or W, X3 is S or D, X4 is Y, H or F, X5 is A or V, X6 is M, I or L, and X7 is S or A; the amino acid sequence of the heavy chain CDR2 includes AISGX8GGSX9X 10 YX 11 X 12 SVKG, where X8 is S or H, X9 is T or H, X 10 is Y, W or F, X 11 A, H, V or L, X 12 is D, A or H; the amino acid sequence of the heavy chain CDR3 includes KX 13 RPX 14 RX 15 X 16 DX 17 , X 13 is Y, F or H, X 14 S, T or H, X 15 V or A, X 16 F, Y, H or W, X 17 is S, A, F, C, H or Y; the amino acid sequence of the light chain CDR1 includes SGDALX 18 DKX 19 X 20 X21 , where X 18 G or H, X 19 Y or F, X 20 A or M, X 21 is S or A; the amino acid sequence of the light chain CDR2 includes X 22 X 23 X 24 X 25 RX 26 X 27 , where X 22 E or Q, X 23 D, H or Q, X 24 is S, A or H, X 25 K, H, R, P or N, X 26 is P, Q, D or N, X 27 is S, D, Y or W; and the amino acid sequence of the light chain CDR3 includes X 28 SNDX 29 DX 30 X 31 W, where X 28 For Q or D, X 29 A, V, I or L, X 30 S or A, X 31 V, Y, W or R.

[0070] In some embodiments, the complementarity determining regions of the antibody or antigen-binding fragment thereof have the following amino acid sequences: the amino acid sequence of the heavy chain CDR region includes SEQ ID NOs: 1-3, 23-64; the amino acid sequence of the light chain CDR region includes SEQ ID NOs: 4-6, 65-109.

[0071] In some embodiments, the complementary determining regions of the antibodies or antigen-binding fragments thereof have amino acid sequences as follows: heavy chain CDR1 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 1, 23-34; heavy chain CDR2 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 2, 35-48; heavy chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 3, 49-64; light chain CDR1 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 4, 65-69; light chain CDR2 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 5, 70-95; and light chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 6, 96-109.

[0072] In some embodiments, the amino acid sequence of the complementarity determining region of the antibody or antigen-binding fragment thereof is selected from at least one of the following groups (1) to (119):

[0073] (1) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 6, respectively;

[0074] (2) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 24, 2, 3, 4, 5, and 6, respectively;

[0075] (3) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 25, 2, 3, 4, 5, and 6, respectively;

[0076] (4) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 26, 2, 3, 4, 5, and 6, respectively;

[0077] (5) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 29, 2, 3, 4, 5, and 6, respectively;

[0078] (6) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 30, 2, 3, 4, 5, and 6, respectively;

[0079] (7) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 31, 2, 3, 4, 5, and 6, respectively;

[0080] (8) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 32, 2, 3, 4, 5, and 6, respectively;

[0081] (9) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 33, 2, 3, 4, 5, and 6, respectively;

[0082] (10) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 37, 3, 4, 5, and 6, respectively;

[0083] (11) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 38, 3, 4, 5, and 6, respectively;

[0084] (12) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 39, 3, 4, 5, and 6, respectively;

[0085] (13) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 40, 3, 4, 5, and 6, respectively;

[0086] (14) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 41, 3, 4, 5, and 6, respectively;

[0087] (15) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 42, 3, 4, 5, and 6, respectively;

[0088] (16) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 43, 3, 4, 5, and 6, respectively;

[0089] (17) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 49, 4, 5, and 6, respectively;

[0090] (18) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 50, 4, 5, and 6, respectively;

[0091] (19) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 51, 4, 5, and 6, respectively;

[0092] (20) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 52, 4, 5, and 6, respectively;

[0093] (21) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 53, 4, 5, and 6, respectively;

[0094] (22) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 54, 4, 5, and 6, respectively;

[0095] (23) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 55, 4, 5, and 6, respectively;

[0096] (24) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 56, 4, 5, and 6, respectively;

[0097] (25) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 57, 4, 5, and 6, respectively;

[0098] (26) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 66, 5, and 6, respectively;

[0099] (27) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 70, and 6, respectively;

[0100] (28) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 71, and 6, respectively;

[0101] (29) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 72, and 6, respectively;

[0102] (30) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 73, and 6, respectively;

[0103] (31) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 74, and 6, respectively;

[0104] (32) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 75, and 6, respectively;

[0105] (33) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 76, and 6, respectively;

[0106] (34) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 77, and 6, respectively;

[0107] (35) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 96, respectively;

[0108] (36) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 3, 4, 5, and 97, respectively;

[0109] (37) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 98, respectively;

[0110] (38) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 3, 4, 5, and 99, respectively; or

[0111] (39) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 100, respectively.

[0112] (40) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 38, 3, 4, 5, and 97, respectively.

[0113] (41) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 2, 57, 4, 5, and 100, respectively.

[0114] (42) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 42, 57, 4, 5, and 99, respectively;

[0115] (43) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 73, and 6, respectively;

[0116] (44) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 57, 4, 73, and 100, respectively;

[0117] (45) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 3, 4, 73, and 100, respectively;

[0118] (46) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 53, 66, 5, and 6, respectively;

[0119] (47) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 4, 5, and 99, respectively;

[0120] (48) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 66, 5, and 99, respectively;

[0121] (49) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 38, 57, 4, 73, and 99, respectively;

[0122] (50) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 73, and 100, respectively;

[0123] (51) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 31, 57, 4, 76, and 100, respectively;

[0124] (52) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 4, 73, and 99, respectively;

[0125] (53) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 53, 4, 73, and 100, respectively;

[0126] (54) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 66, 5, and 99, respectively;

[0127] (55) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 39, 53, 66, 5, and 100, respectively;

[0128] (56) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 73, and 104, respectively;

[0129] (57) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 66, 73, and 104, respectively;

[0130] (58) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 46, 57, 4, 73, and 104, respectively;

[0131] (59) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 64, 4, 73, and 104, respectively;

[0132] (60) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 63, 4, 73, and 104, respectively;

[0133] (61) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 57, 4, 85, and 100, respectively;

[0134] (62) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 31, 42, 57, 4, 87, and 100, respectively;

[0135] (63) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 57, 4, 73, and 106, respectively;

[0136] (64) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 63, 66, 73, and 100, respectively;

[0137] (65) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 57, 4, 85, and 104, respectively;

[0138] (66) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 25, 42, 63, 4, 73, and 104, respectively;

[0139] (67) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 29, 42, 62, 4, 87, and 100, respectively;

[0140] (68) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 32, 42, 63, 4, 73, and 105, respectively;

[0141] (69) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 63, 4, 85, and 100, respectively;

[0142] (70) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 47, 63, 66, 73, and 100, respectively;

[0143] (71) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 63, 66, 73, and 104, respectively;

[0144] (72) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 63, 66, 73, and 104, respectively;

[0145] (73) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 63, 4, 85, and 104, respectively;

[0146] (74) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 64, 4, 87, and 104, respectively;

[0147] (75) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 48, 63, 66, 85, and 100, respectively;

[0148] (76) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 43, 63, 66, 73, and 106, respectively;

[0149] (77) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 88, and 100, respectively;

[0150] (78) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 89, and 100, respectively;

[0151] (79) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 4, 90, and 100, respectively;

[0152] (80) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 92, and 100, respectively;

[0153] (81) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 42, 57, 4, 91, and 100, respectively;

[0154] (82) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 94, and 100, respectively;

[0155] (83) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 93, and 100, respectively;

[0156] (84) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 93, and 109, respectively;

[0157] (85) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 23, 42, 57, 4, 73, and 100, respectively;

[0158] (86) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 29, 42, 57, 4, 73, and 100, respectively;

[0159] (87) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 57, 4, 73, and 108, respectively;

[0160] (88) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 73, and 107, respectively;

[0161] (89) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 73, and 109, respectively;

[0162] (90) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 73, and 100, respectively;

[0163] (91) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 69, 73, and 100, respectively;

[0164] (92) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 73, and 109, respectively;

[0165] (93) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 93, and 109, respectively;

[0166] (94) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 86, and 100, respectively;

[0167] (95) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 86, and 100, respectively;

[0168] (96) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 67, 73, and 108, respectively;

[0169] (97) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 95, and 108, respectively;

[0170] (98) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 57, 4, 94, and 108, respectively;

[0171] (99) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 100, respectively;

[0172] (100) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 73, and 100, respectively;

[0173] (101) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 68, 73, and 100, respectively;

[0174] (102) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 108, respectively;

[0175] (103) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 107, respectively;

[0176] (104) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 109, respectively;

[0177] (105) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 69, 73, and 100, respectively;

[0178] (106) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 73, and 109, respectively;

[0179] (107) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 86, and 100, respectively;

[0180] (108) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 4, 93, and 109, respectively;

[0181] (109) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 86, and 100, respectively;

[0182] (110) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 34, 42, 57, 65, 73, and 100, respectively;

[0183] (111) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 44, 57, 68, 73, and 100, respectively;

[0184] (112) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 45, 57, 65, 73, and 100, respectively;

[0185] (113) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 45, 57, 69, 73, and 100, respectively;

[0186] (114) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 45, 57, 4, 86, and 100, respectively;

[0187] (115) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 28, 42, 57, 4, 73, and 108, respectively;

[0188] (116) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 27, 42, 59, 4, 73, and 108, respectively;

[0189] (117) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 58, 4, 73, and 108, respectively;

[0190] (118) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 60, 4, 73, and 108, respectively;

[0191] (119) The amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 61, 4, 73, and 108, respectively.

[0192] In some embodiments, the antibody or antigen-binding fragment thereof further comprises a framework region (FR region). Suitable FR region sequences are known in the art, and any suitable FR region sequence can be used in the present application. In some embodiments, the FR region of the heavy chain comprises the FR region of the heavy chain variable region domain set forth in SEQ ID NO: 7. In some embodiments, the FR region of the light chain comprises the FR region of the light chain variable region domain set forth in SEQ ID NO: 8.

[0193] In some embodiments, the heavy chain variable region of the antibody or antigen-binding fragment thereof has an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the sequence shown in SEQ ID NO: 7, and the light chain variable region has an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the sequence shown in SEQ ID NO: 8. In some preferred embodiments, the heavy chain variable region of the antibody or antigen-binding fragment thereof has the sequence shown in SEQ ID NO: 7, and the light chain variable region has the sequence shown in SEQ ID NO: 8.

[0194] In some embodiments, the heavy chain of the antibody or antigen-binding fragment thereof is an amino acid sequence obtained by mutation of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 positions in the sequence shown in SEQ ID NO: 11. In some embodiments, the mutation is selected from insertion, deletion and / or substitution. In some preferred embodiments, the mutation is substitution. In some embodiments, the substituted amino acid position is selected from at least one of positions 27, 29, 31, 32, 33, 34, 35, 54, 58, 59, 61, 62, 100, 103, 105, 106, and 108 of the sequence shown in SEQ ID NO: 11. In some preferred embodiments, the replaced amino acid site is selected from at least one of F27Y, F29H, F29Y, F29W, S31D, Y32F, Y32H, A33V, M34I, M34L, S35A, S54H, T58H, Y59F, Y59W, A61H, A61V, A61L, D62A, D62H, Y100H, Y100F, S103H, S103T, V105A, F106H, F106Y, F106W, S108A, S108C, S108F, S108H, and S108Y.

[0195] In some embodiments, the light chain of the antibody or antigen-binding fragment thereof is an amino acid sequence obtained by mutation of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 positions in the sequence shown in SEQ ID NO: 12. In some embodiments, the mutation is selected from insertion, deletion and / or substitution. In some preferred embodiments, the mutation is substitution. In some embodiments, the substituted amino acid position is selected from at least one of positions 28, 31, 32, 33, 49, 50, 51, 52, 54, 55, 88, 92, 94, and 95 of the sequence shown in SEQ ID NO: 12. In some preferred embodiments, the replaced amino acid site is selected from at least one of G28H, Y31F, A32M, S33A, E49Q, D50H, D50Q, S51A, S51H, K52H, K52R, K52P, K52N, P54D, P54N, P54Q, S55D, S55W, S55Y, Q88D, A92V, A92L, A92I, S94A, V95R, V95W, and V95Y.

[0196] In some embodiments, the antibody or antigen-binding fragment thereof further comprises a constant region. Suitable constant region sequences are known in the art, and any suitable constant region sequence can be used in the present application. In some preferred embodiments, the amino acid sequence of the heavy chain constant region is as shown in SEQ ID NO: 9, and the amino acid sequence of the light chain constant region is as shown in SEQ ID NO: 10.

[0197] In some embodiments, the antibody or antigen-binding fragment thereof may further optionally comprise a signal peptide. When a prokaryotic expression system is used to express and purify the antibody or antigen-binding fragment thereof, suitable signal peptide sequences are known in the art, and any suitable signal peptide sequence may be used in the present application. In some embodiments, the heavy chain comprises a heavy chain signal peptide as shown in SEQ ID NO: 13, and the light chain comprises a light chain signal peptide as shown in SEQ ID NO: 14. In some preferred embodiments, the heavy chain comprising the signal peptide has the sequence shown in SEQ ID NO: 15, and the light chain comprising the signal peptide has the sequence shown in SEQ ID NO: 16.

[0198] In some embodiments, the antibody or antigen-binding fragment thereof does not contain a signal peptide. When a eukaryotic expression system is used to express and purify the antibody or antigen-binding fragment thereof, the heavy chain and light chain may not contain a signal peptide. In some preferred embodiments, the heavy chain that does not contain a signal peptide has the sequence shown in SEQ ID NO: 11, and the light chain that does not contain a signal peptide has the sequence shown in SEQ ID NO: 12.

[0199] In some embodiments, the antibody or antigen-binding fragment thereof includes but is not limited to a single-chain antibody, a single-domain antibody, a diabody, a triabody, a disulfide-stabilized antibody, a nanobody, a Fab fragment, a Fab' fragment, a (Fab')2 fragment, a minibody, a monoclonal antibody, a polyclonal antibody, a monospecific antibody, a bispecific antibody, a multispecific antibody, a monovalent antibody, a multivalent antibody, a chimeric antibody, a fusion protein comprising an antigen-binding site of an antibody, or a whole antibody immunoglobulin IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD.

[0200] Polynucleotides, recombinant vectors, host cells

[0201] According to one embodiment of the present application, a polynucleotide is also provided, which encodes the antibody or antigen-binding fragment thereof described in the present application.

[0202] According to one embodiment of the present application, a recombinant vector is also provided, comprising the polynucleotide described herein. Recombinant vectors suitable for expressing the antibodies or antigen-binding fragments described herein are well known in the art and are not limited here.

[0203] In some embodiments, the recombinant vector further comprises a promoter. The promoter can be any suitable promoter sequence, i.e., a nucleic acid sequence that is recognized by the host cell expressing the nucleic acid sequence. The promoter sequence contains transcriptional regulatory sequences that mediate expression of the antibody or antigen-binding fragment thereof. The promoter can be any nucleic acid sequence that is transcriptionally active in the selected host cell, including mutant, truncated, and hybrid promoters, and can be derived from a gene encoding an extracellular or intracellular protein or polypeptide that is homologous or heterologous to the host cell.

[0204] The recombinant vector of the present application can be constructed using methods well known in the art. For example, appropriate restriction sites can be added to both ends of the polynucleotide of the present application according to the restriction sites contained in the backbone vector used, and then loaded into the backbone vector.

[0205] According to one embodiment of the present application, a host cell is also provided, comprising the polynucleotides described herein and / or the recombinant vectors described herein. The host cell can be selected based on the type of expression vector. The polynucleotides and / or the expression vectors can be delivered into the host cell using any suitable means known in the art, without limitation.

[0206] Pharmaceutical compositions, kits

[0207] According to one embodiment of the present application, a pharmaceutical composition is further provided, characterized in that the pharmaceutical composition comprises the antibody or antigen-binding fragment thereof described in the present application.

[0208] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically or physiologically acceptable carrier. The carrier can be any compatible, physiologically acceptable, non-toxic substance suitable for delivering the polypeptide, polynucleotide, or recombinant vector provided herein into a mammal (e.g., a human).

[0209] The term "pharmaceutically acceptable carrier" refers to a carrier, diluent, or adjuvant used in the formulation or administration of the polypeptides, polynucleotides, or recombinant vectors provided herein, which is not itself an essential active ingredient and is not unduly toxic upon administration. Suitable pharmaceutically acceptable carriers are well known to those of ordinary skill in the art.

[0210] Among them, "physiologically acceptable carrier" refers to a carrier, diluent, or adjuvant that does not cause significant irritation to an organism and does not eliminate the pharmaceutical activity and properties of the administered polypeptide, polynucleotide, or recombinant vector provided herein. Suitable physiologically acceptable carriers are also well known to those of ordinary skill in the art.

[0211] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient. In some embodiments, the excipient comprises at least one of a solubilizer, a disintegrant, a wetting agent, a stabilizer, a thickener, a diluent, a buffer, and a flavoring agent.

[0212] In some non-limiting embodiments, the carriers and / or excipients used in the pharmaceutical compositions of the present application may include, for example, liquid, gel or solid carriers, aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, suspending agents, dispersants, chelating agents, diluents, adjuvants, excipients or non-toxic auxiliary substances, other components known in the art, or various combinations thereof.

[0213] According to one embodiment of the present application, a kit is also provided, characterized in that the kit comprises the antibody or antigen-binding fragment thereof described in the present application.

[0214] Multispecific antibodies and antibody conjugates

[0215] According to one embodiment of the present application, a multispecific antibody is also provided, which comprises the antibody or antigen-binding fragment thereof described in the present application, and further comprises one or more second antibodies or antigen-binding fragments thereof that specifically bind to other antigens.

[0216] In some embodiments, the second antibody or antigen-binding fragment thereof is selected from a full-length antibody, Fab, Fab", (Fab")2, Fv, scFv, scFv-scFv, minibody, diabody or sdAb.

[0217] Those skilled in the art can select a suitable second antibody or antigen-binding fragment thereof as needed, and use methods known in the art to couple the antibody or antigen-binding fragment provided herein with the second antibody or antigen-binding fragment thereof to form a multispecific antibody.

[0218] According to one embodiment of the present application, an antibody conjugate is also provided, which comprises the antibody or antigen-binding fragment described in the present application and a second functional structure, wherein the second functional structure is selected from Fc, a radioactive isotope, a structural portion that extends half-life, a detectable marker and a drug.

[0219] Suitable Fc, radioactive isotopes, structural parts that extend half-life, detectable labels and drugs are known in the art. Those skilled in the art can select a suitable second functional structure as needed and couple the antibody or antigen-binding fragment provided by the present application to the second functional structure using methods known in the art.

[0220] In some embodiments, the antibody conjugate comprises the anti-S100A8 / A9 antibody provided herein and Fc. The term "Fc" is used to define the C-terminal region of an immunoglobulin heavy chain, including native Fc and variant Fc. "Native Fc" refers to a molecule or sequence comprising a non-antigen binding fragment produced by digestion of an intact antibody, whether in monomeric or multimeric form. The immunoglobulin source from which native Fc is produced is preferably human. Native Fc fragments are composed of monomeric polypeptides that can be linked into dimer or multimeric forms by covalent linkages (e.g., disulfide bonds) and non-covalent linkages. Depending on the class (e.g., IgG, IgA, IgE, IgD, IgM) or subtype (e.g., IgG1, IgG2, IgG3, IgA1, IgGA2), native Fc molecules have 1-4 intermolecular disulfide bonds between monomeric subunits. An example of a native Fc is a disulfide-linked dimer produced by digesting IgG with papain (see Ellison et al. (1982), Nucleic Acids Res. 10: 4071-9). The term "native Fc" as used herein generally refers to monomeric, dimeric and multimeric forms. "Variant Fc" refers to an amino acid sequence that differs from the amino acid sequence of a "native" or "wild-type" Fc due to at least one "amino acid modification" as defined herein, also referred to as an "Fc variant". Therefore, "Fc" also includes single-chain Fc (scFc), that is, a single-chain Fc composed of two Fc monomers connected by a polypeptide linker, which can naturally fold into a functional dimeric Fc region. In one embodiment, the Fc is preferably the Fc of a human immunoglobulin, more preferably the Fc of human IgG1.

[0221] In some embodiments, the antibody conjugate comprises the anti-S100A8 / A9 antibody provided herein and a drug to form an antibody-drug conjugate (ADC), wherein the drug is selected from a cytotoxin and an immunomodulator. In some embodiments, examples of cytotoxins include, but are not limited to, methotrexate, aminopterin, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, dacarbazine, mechlorethamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), 1-methylnitrosourea, cyclophosphamide, mechlorethamine, busulfan, dibromomannitol, streptozocin, mitomycin, cis-dichlorodiamine platinum (II) (DDP), cisplatin. , carboplatin, doxycycline, doxorubicin, detoxorubicin, camiomycin, idarubicin, epirubicin, mitoxantrone, actinomycin D, bleomycin, calicheamicin, mithramycin, anthramycin (AMC), vincristine, vinblastine, paclitaxel, ricin, Pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, teniposide, colchicine, dihydroxyanthraquinone, 1-dehydrotestosterone, glucocorticoids In some embodiments, examples of immunomodulators include, but are not limited to, ganciclovir, etanercept, tacrolimus, sirolimus, voriconazole, cyclosporine, rapamycin, cyclophosphamide, azathioprine, mycophenolate mofetil, methotrexate, glucocorticoids and their analogs, cytokines, stem cell growth factors, lymphotoxins, tumor necrosis factor (TNF), hematopoietic factors, interleukins (e.g., IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, and IL-21), colony stimulating factors (e.g., G-CSF and GM-CSF), interferons (e.g., interferon-α, interferon-β, and interferon-γ), erythropoietin, and thrombopoietin, or a combination thereof.

[0222] In some embodiments, an antibody conjugate comprises an anti-S100A8 / A9 antibody provided herein and a radioactive isotope to form a radionuclide drug conjugate (RDC). Examples of radioactive isotopes that can be used in the present invention include, but are not limited to, At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, 99mTc, 123I, 18F, and 68Ga.

[0223] In some embodiments, an antibody conjugate comprises an anti-S100A8 / A9 antibody provided herein and a half-life-extending moiety to improve the half-life of the anti-S100A8 / A9 antibody. The half-life-extending moiety is selected from the group consisting of an albumin binding structure, a transferrin binding structure, a polyethylene glycol molecule, a recombinant polyethylene glycol molecule, human serum albumin, a fragment of human serum albumin, and an albumin polypeptide that binds to human serum albumin.

[0224] In some embodiments, the antibody conjugate comprises an anti-S100A8 / A9 antibody provided herein and a detectable marker selected from a fluorophore, a chemiluminescent compound, a bioluminescent compound, an enzyme, an antibiotic resistance gene, and a contrast agent. The term "detectable marker" as used herein refers to a compound that produces a detectable signal. For example, the detectable marker can be an MRI contrast agent, a scintigraphy contrast agent, an X-ray imaging contrast agent, an ultrasound contrast agent, or an optical imaging contrast agent. Examples of detectable markers include fluorophores (such as fluorescein, Alexa, or cyanine), chemiluminescent compounds (such as luminol), bioluminescent compounds (such as luciferase or alkaline phosphatase), enzymes (such as horseradish peroxidase, glucose-6-phosphatase, β-galactosidase), antibiotics (such as kanamycin, ampicillin, chloramphenicol, tetracycline, etc.) resistance genes, and contrast agents (such as nanoparticles or gadolinium). Those skilled in the art can select a suitable detectable marker based on the detection system used.

[0225] use

[0226] According to one embodiment of the present application, there is also provided use of the antibody or antigen-binding fragment thereof, the multispecific antibody, and / or the antibody conjugate described herein in the preparation of drugs, reagents, and kits for detecting, preventing, or treating diseases.

[0227] In some embodiments, the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers of tetramers or greater of S100A8 and S100A9. As known in the art, there are a variety of diseases or non-disease symptoms accompanied by increased expression levels of S100A8 / S100A9 oligomers, dimers, tetramers or multimers of more than four polymers, including but not limited to sepsis (Am J Respir Crit Care Med, 2017, 196(3):315-327), autoimmune diseases (Autoimmun Rev, 2023, 22(5):103295), acute lung injury (Blood, 2022, 140(24):2626-2643), acute pancreatitis (Dis Markers, 2018, 2018:6457347), acute kidney injury (Adv Sci (Weinh), 2022, 9(12):e2103675), tumors (Sci Transl Med, 2020, 12(572):eabb5817), organ ischemia / reperfusion injury (Circulation, 2019, 140(9):751-764; Kidney Int, 2015, 87(1):85-94), atherosclerosis-related cardiocerebral disease (Pharmacol Res, 2020, 161:105212), these disclosures are incorporated herein by reference in their entirety. In some preferred embodiments, the disease includes myocardial infarction and myocardial ischemia / reperfusion injury.

[0228] According to one embodiment of the present application, there is also provided the use of the antibody or antigen-binding fragment thereof, the multispecific antibody, and / or the antibody conjugate described herein in the preparation of drugs, reagents, and kits for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function.

[0229] In the purposes described herein, the dosage of the antibody or antigen-binding fragment provided herein may depend on several factors, including the severity and responsiveness of the symptoms, the route of administration, the duration of treatment (a few days to several months to several years), and the time to improvement of the symptoms. Those skilled in the art can adjust the dosage regimen to provide a therapeutic response according to the patient's specific circumstances. For example, a single administration may be performed, several separate dosages may be administered within a predetermined time period, or the dosage may be reduced or increased as indicated by the treatment situation. The specification of the dosage is determined by the specific therapeutic effect to be achieved. The dosage value may also vary with the type and severity of the condition to be alleviated. For any particular subject, the specific dosage regimen may be adjusted over time according to individual needs and the professional judgment of the treating clinician.

[0230] Detection, prevention, or treatment of diseases

[0231] According to one embodiment of the present application, a method for detecting, preventing or treating a disease is also provided, comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof described herein, the pharmaceutical composition described herein, the kit described herein, the multispecific antibody described herein, and / or the antibody conjugate described herein.

[0232] In some embodiments, the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers of tetramers or greater of S100A8 and S100A9. As known in the art, there are a variety of diseases or non-disease symptoms accompanied by increased expression levels of S100A8 / S100A9 oligomers, dimers, tetramers or multimers of more than four polymers, including but not limited to sepsis (Am J Respir Crit Care Med, 2017, 196(3):315-327), autoimmune diseases (Autoimmun Rev, 2023, 22(5):103295), acute lung injury (Blood, 2022, 140(24):2626-2643), acute pancreatitis (Dis Markers, 2018, 2018:6457347), acute kidney injury (Adv Sci (Weinh), 2022, 9(12):e2103675), tumors (Sci Transl Med, 2020, 12(572):eabb5817), organ ischemia / reperfusion injury (Circulation, 2019, 140(9):751-764; Kidney Int, 2015, 87(1):85-94), and atherosclerosis-related cardiocerebral diseases (Pharmacol Res, 2020, 161:105212), the disclosures of which are incorporated herein by reference in their entirety. In some preferred embodiments, the diseases include ischemic organ damage and organ ischemia / reperfusion injury associated with S100A8 / A9, S100A8, or S100A9.

[0233] According to one embodiment of the present application, a method for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function is also provided, the method comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof described herein, the pharmaceutical composition described herein, the kit described herein, the multispecific antibody described herein, and / or the antibody conjugate described herein.

[0234] In the methods described herein, the dosage of the antibody or antigen-binding fragment provided herein may depend on several factors, including the severity and responsiveness of the symptoms, route of administration, treatment time (several days to several months to several years), and the time to improvement of the symptoms. Those skilled in the art can adjust the dosage regimen to provide a therapeutic response according to the patient's specific circumstances. For example, a single administration may be performed, several separate dosages may be administered within a predetermined time period, or the dosage may be reduced or increased as indicated by the treatment situation. The specification of the dosage is determined by the specific therapeutic effect to be achieved. The dosage value may also vary with the type and severity of the condition to be alleviated. For any particular subject, the specific dosage regimen may be adjusted over time according to individual needs and the professional judgment of the treating clinician.

[0235] The various embodiments and preferences disclosed above can be combined with each other (as long as they are not inherently contradictory to each other), and the various embodiments formed by such combination are all considered to be part of the disclosure of this application.

[0236] The following description will be made of exemplary embodiments of the present application in conjunction with the accompanying drawings, including various details of the embodiments of the present application to facilitate understanding. It should be understood that they are considered to be merely exemplary and are in no way intended to limit the scope of protection of the present application. The scope of protection of the present application is defined solely by the claims. Therefore, it should be appreciated by those skilled in the art that various changes and modifications may be made to the embodiments described herein without departing from the scope of the present application. Similarly, for the sake of clarity and conciseness, descriptions of well-known functions and structures are omitted in the following description.

[0237] Example

[0238] If no specific techniques or conditions are specified in this example, the experiments were carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially.

[0239] Example 1: Preparation of prokaryotic recombinant antigens from different species

[0240] S100A8 (Sino Biological, Cat. No. HG11138-CH, Uniprot: P05109, amino acid sequence shown in SEQ ID No: 110, encoding nucleic acid sequence shown in SEQ ID No: 111) and S100A9 (Sino Biological, Cat. No. HG11145-CH, Uniprot: P06702, amino acid sequence shown in SEQ ID No: 112, encoding nucleic acid sequence shown in SEQ ID No: 113) gene fragments were purchased for protein expression in Escherichia coli. The gene fragments were cloned into the pET21a vector (Novagen) digested with NdeI and BamHI to construct the pET21-S100A8 or pET21-S100A9 vectors. The co-expression vector pET21-S100A8-S100A9 was used for co-expression experiments. Similar methods were used to construct and produce recombinant proteins from different species, including human S100A8 / A9 (hS100A8 / A9), mouse S100A8 / A9 (mS100A8 / A9), and monkey S100A8 / A9 (cS100A8 / A9). A His-tag was engineered at the C-terminus of the recombinant protein for purification. Escherichia coli BL21(DE3)T7 expression cells were transformed with the expression plasmids. Colonies were inoculated in 20 mL of LB supplemented with 100 μg / mL ampicillin and cultured in a shake flask at 37°C for 2 hours. Twenty mL of the preculture was inoculated into 800 mL of LB supplemented with 100 μg / mL ampicillin. When the cell density reached an OD600 of 0.5, 0.5 mM isopropyl β-d-1-thiogalactopyranoside (IPTG) was added for induction for 3 hours. The E. coli cells were harvested by centrifugation and washed with 0.15 M NaCl. The cells were resuspended in 80 mL of 50 mM Tris-HCl buffer (pH 7.5) containing 50 mM NaCl and 5 mM MgSO4 and disrupted by sonication on ice. The supernatant collected by centrifugation after sonication was purified using metal chelate affinity chromatography (Ni-NTA) to obtain a fusion recombinant protein with a His-tag tag. The purified target protein was dialyzed and replaced with PBS, then aliquoted and stored at -80°C. The recombinant protein was then depyrogenated according to the needs of different experiments, and the pyrogen-free recombinant protein was then aliquoted and stored at -80°C.

[0241] Example 2: Preparation and activity detection of anti-S100A8 / A9 antibodies

[0242] 2.1 Screening of anti-hS100A8 / A9 antibodies using phage antibody library

[0243] Immunotubes were coated with hS100A8 / A9-His (hS100A8 / A9) antigen prepared in Example 1 at 20 μg / mL, 1 mL / tube, and incubated at 4°C overnight. The following day, the tubes were washed with PBS 4 times, 3 minutes each time, and then blocked with 1.5 mL of 2% BSA at 37°C for 2 hours. A phage library displaying single-chain antibodies (constructed by our company) was blocked with blocking solution (2% BSA, 0.1% Tween 20) at 37°C for 30 minutes and then added to the immunotubes. The dosage was 10 9 -10 12 / tube, bind at 37℃ for 1 hour. After thorough washing with PBST and PBS, add 1mL of 0.2mol / L glycine-HCl (pH2.2) for elution, shake at room temperature for 10 minutes, and immediately neutralize with 1mol / L Tris (pH 8.8) to a pH of approximately 7.0. Infect the neutralized phage with XL1-Blue activated bacteria in the logarithmic growth phase, incubate in a 37℃ incubator for 20-30 minutes, and then culture on a shaker for 30 minutes at 37℃ at 150rpm. Then, take 1% and 0.1% of the bacterial solution for plate counting. Centrifuge the remaining bacterial solution at 4000rpm for 10 minutes, discard the supernatant, spread the bacteria on 2YTCG solid plates, and then culture overnight at 37℃. The next day, cells were collected from the plates and inoculated into 2YTCG liquid medium with shaking until the logarithmic growth phase. The cells were then infected with M13KO7 and allowed to stand at room temperature for 15-30 minutes. The cells were then incubated at 37°C / 150 rpm for 1 hour. 50 μg / mL kanamycin was then added and the cells were incubated at 30°C overnight. The next day, phage were purified by precipitation using PEG8000 / NaCl. These phage were then used for the next round of screening. Three rounds of phage library enrichment and screening were performed.

[0244] 2.2 Monoclonal Identification of Anti-hS100A8 / A9 Single-chain Antibody

[0245] After three rounds of phage screening, single colonies were selected and inoculated into 200 μL of 2YTCTG medium. Cultured at 37°C and 220 rpm until the logarithmic growth phase, and then 10 8Helper phage M13KO7 was infected at room temperature for 20 minutes, followed by incubation at 37°C at 150 rpm for 1 hour. An equal volume of 2×YTCTKI (kanamycin 50 μg / mL, IPTG 0.2 mmol / L) was then added and incubated at 30°C at 200 rpm overnight. The next day, the supernatant was collected by centrifugation, and BSA was added to a final concentration of 2%, followed by Tween-20 to a final concentration of 0.1%. The cells were incubated at 37°C for 15 minutes for ELISA analysis. A 96-well ELISA plate was coated with hS100A8 / A9 antigen at 200 ng / well at 4°C overnight. The next day, the plate was blocked with PBST-4% milk (250 μL / well) at 37°C for 2 hours. The blocking solution was discarded, and the blocked phage antibody was added. The plate was incubated at 37°C for 1 hour. The solution was discarded, and the plate was washed three times with PBST. Anti-M13 Antibody (HRP) (Sino Biological, Cat: 11973-MM05T-H) was diluted in blocking buffer and added to the ELISA plate. The plate was incubated at 37°C for 30 minutes. After washing three times with PBST, OPD substrate was added to the plate and color was developed at room temperature for approximately 10 minutes. The color development was terminated with 2M H2SO4. Absorbance was measured using a microplate reader.

[0246] 2.3 Conversion of single-chain antibodies to full antibodies and expression and purification

[0247] The anti-hS100A8 / A9 single-chain antibodies obtained after screening were used to prepare full-length antibodies. Primers LF (corresponding nucleotide sequence shown in SEQ ID No: 19) and LR (corresponding nucleotide sequence shown in SEQ ID No: 20) were used to amplify the VL variable region gene; primers HF (corresponding nucleotide sequence shown in SEQ ID No: 21) and HR (corresponding nucleotide sequence shown in SEQ ID No: 22) were used to amplify the VH variable region gene. Vectors pABL and pABG4 were used to clone the VL and VH variable region genes, respectively. VL and VH were cloned into the vectors using homologous recombination methods. After transformation into Escherichia coli Top10, single clones were selected and the recombinant plasmids were sequenced and identified to select the correct heavy and light chain expression vectors for the fully human antibody. After plasmid extraction, the heavy chain and light chain plasmids were transfected into 293-T cells at a molar ratio of 1:1 for transient expression of the full antibody. The expression supernatant was purified by Protein A affinity chromatography, and the purified full antibody was identified by electrophoresis and packaged for storage for subsequent experimental identification. The antibody was pyrogen-free according to the needs of different experiments.

[0248] 2.4 Binding activity assay of anti-S100A8 / A9 mAbs

[0249] The binding activity of four candidate monoclonal antibodies to the hS100A8 / A9 antigen was analyzed by ELISA. The expressed and purified monoclonal antibodies were coated onto 96-well plates (0.5 μg / mL, 50 μL / well, overnight at 4°C). The affinity of the monoclonal antibodies for hS100A8 / A9 antigen was measured at various dilutions. The starting concentration of hS100A8 / A9 antigen was 100 μg / mL, and a 5-fold or 3-fold serial dilution was performed, resulting in eight dilutions. The HRP-conjugated secondary antibody used was an HRP-conjugated 6×His-Tag monoclonal antibody (Proteintech, Cat No: HRP-66005) (see Figure 1). Sequence information is shown in Table 1.

[0250] Table 1 Antibody related sequences

[0251] Note: In the amino acid sequences of the heavy and light chains above, the underlined and double-lined portion at the downstream of the sequence represents the constant region sequence; the bolded portion at the middle of the sequence represents the complementarity-determining region (CDR) sequence.

[0252] 2.5 Activity Analysis of Anti-S100A8 / A9 Monoclonal Antibodies

[0253] 2.5.1 Reichert 4SPR Detection of Anti-S100A8 / A9 Monoclonal Antibody Affinity

[0254] Anti-S100A8 / A9 monoclonal antibodies were covalently coupled to an SR7000 GOLD SENSOR SLIDE biosensor chip (Reichert, PARTNO: 13206066). When target molecules flowed across the chip surface, they were captured by the anti-S100A8 / A9 monoclonal antibodies on the chip. The reaction signals were detected in real time using a Reichert 4SPR instrument, thereby obtaining antigen-antibody binding and dissociation curves, which were then fitted to obtain affinity values. The present application further tested the affinity of the screened anti-S100A8 / A9 monoclonal antibodies to the human heterodimeric antigen hS100A8 / A9 and the monkey heterodimeric antigen cS100A8 / A9 using a Reichert 4 SPR. The results are shown in Table 2 below. The results show that the anti-S100A8 / A9 monoclonal antibodies screened in the present application have the strongest affinity to hS100A8 / A9, and have weaker affinity to cS100A8 / A9 than hS100A8 / A9.

[0255] Table 2 Affinity test results

[0256] 2.5.2 Specificity of anti-S100A8 / A9 mAbs and proteins in the same family

[0257] The binding specificity of anti-S100A8 / A9 monoclonal antibodies to the same family proteins was detected by ELISA. Human S100 family proteins (S100A1, Uniprot: P23297; S100A2, Uniprot: P29034; S100A3, Uniprot: P33764; S100A4, Uniprot: P26447; S100A5, Uniprot: P33763; S100A6, Uniprot: P06703; S100A7, Uniprot: P31151; S100A8 / A9 monoclonal antibodies to the same family proteins) were coated on 96-well plates. 100A8, Uniprot: P05109; S100A9, Uniprot: P06702; S100A10, Uniprot: P60903; S100A11, Uniprot: P3194 9; S100A12, Uniprot: P80511; S100A13, Uniprot: Q99584; S100A14, Uniprot: Q9HCY8; S100A15, Uniprot: Q86SG5; S100A16, Uniprot: Q96FQ6; S100A7L2, Uniprot: Q5SY68; S100B, Uniprot: P04271; S100G, Uniprot: P29377; S100P, Uniprot: P25815; S100Z, Uniprot: Q8WXG8; S100A8 / A9) (0.5 μg / mL, 50 μL / well, coated overnight at 4°C), the next day the liquid in the plate was discarded, and the plate was blocked with blocking solution (4% milk-PBS) at 37°C for 1 hour, and anti-S100A8 / A9 monoclonal antibodies were added at a concentration of 20 μg / mL, and bound to the plate at 37°C for 1 hour, then washed four times with PBST, and then horseradish enzyme-labeled goat anti-human IgG (Zhongshan Jinqiao, ZB-2304) was added. ELISA results showed that the anti-S100A8 / A9 monoclonal antibody prepared in the present application specifically bound to hS100A8 / A9 and had no significant binding to other S100 family proteins ( FIG. 2 ).

[0258] 2.5.3 Specificity of anti-S100A8 / A9 mAbs and other unrelated proteins

[0259] The binding specificity of anti-S100A8 / A9 mAbs to unrelated proteins was detected by ELISA. 96-well plates were coated with 28 other random unrelated proteins (HT, Uniprot: P07911; AFP, Uniprot: P02771; Znt8Nc, Uniprot: Q8IWU4; GAD65, Uniprot: Q05329; ompH, Uniprot: A9KC33; coml, Uniprot: Q57333; nCov-S1-591, Uniprot: P0DTC2; adaAT, Uniprot: L0BY86; hFAB P, Uniprot: P05413; TSLP, Uniprot: Q969D9; Rat-GP73, Uniprot: D4AEL2; hGP73, Uniprot: Q8NBJ4; CKMB, Uniprot: P 06732+P12277; G-Trx, Uniprot: P01350+P0AA25; AfoleA, Uniprot: G1XA82; plasma, Uniprot: P26948; β-T, Uniprot : P05067+P0AA25; IL-1β, Uniprot: P01584; IL-2, Uniprot: P60568; IL2Ra, Uniprot: P01589; IL-4, Uniprot: P05112 ; IL-5, Uniprot: P05113; IL5D2, Uniprot: P05113; IL-8, Uniprot: P10145; IL-8D2, Uniprot: P10145; IL-10, Unipro t: P22301; IL-10D2, Uniprot: P22301; IL-17A, Uniprot: Q16552; (0.5 μg / mL, 50 μL / well, coated overnight at 4°C). The next day, the liquid in the plate was discarded and blocked with blocking buffer (4% milk-PBS) at 37°C for 1 hour. Anti-S100A8 / A9 monoclonal antibodies were added at a concentration of 20 μg / mL and allowed to bind at 37°C for 1 hour. The plates were then washed four times with PBST, and horseradish enzyme-conjugated goat anti-human IgG (Zhongshan Jinqiao, ZB-2304) was added. ELISA results showed that the anti-S100A8 / A9 monoclonal antibodies prepared in this application specifically bound to hS100A8 / A9 and showed no significant binding to random unrelated proteins (Figure 3).

[0260] 2.5.4 Species Specificity Analysis of Anti-S100A8 / A9 Monoclonal Antibodies

[0261] The species specificity of anti-S100A8 / A9 mAbs was tested by ELISA. 96-well plates were coated with anti-S100A8 / A9 mAbs (0.5 μg / mL, 50 μL / well, overnight at 4°C). The next day, the plates were discarded and blocked with blocking buffer (4% milk-PBS) at 37°C for 1 hour. Three species of S100A8 / A9 antigens (human heterodimeric antigen hS100A8 / A9, mouse heterodimeric antigen mS100A8 / A9, and monkey heterodimeric antigen cS100A8 / A9) were added at a starting concentration of 20 μg / mL. Five-fold gradients were set for a total of eight steps. The plates were allowed to bind for 1 hour at 37°C and then washed four times with PBST. The secondary antibody used was HRP-conjugated 6×His, His-Tag Monoclonal antibody (Proteintech, Cat No: HRP-66005). ELISA results showed that the anti-S100A8 / A9 monoclonal antibodies screened in this application had the strongest affinity for hS100A8 / A9, followed by cS100A8 / A9, while the anti-S100A8 / A9 monoclonal antibodies did not significantly bind to mS100A8 / A9 (Figure 4).

[0262] 2.5.5 Stability Analysis of Anti-S100A8 / A9 Monoclonal Antibodies in Serum and PBS

[0263] The present application further analyzes the stability of anti-S100A8 / A9 monoclonal antibodies in PBS and C57BL / 6 mouse serum (m-Serum). The specific implementation method is as follows: Take the filter-sterilized anti-S100A8 / A9 monoclonal antibody samples and dilute them to 100 μg / mL with sterile PBS and mouse serum, respectively, with a volume of 200 μL / vial, and place them in a 37°C incubator for 0 days, 1 day, 4 days, 6 days, 8 days, 10 days, 13 days, 15 days, 18 days, 21 days, and 25 days. After 25 days, the binding of the samples in serum and PBS to the hS100A8 / A9 antigen was analyzed by ELISA (Figures 5A and 5B). The results show that the hS100A8 / A9 antibody obtained in the present application has good antigen binding stability in both C57BL / 6 mouse serum and PBS.

[0264] Example 3: Epitope Analysis of Anti-S100A8 / A9 Monoclonal Antibodies Binding to hS100A8 / A9

[0265] The binding epitopes of antigens and antibodies were analyzed by cryo-electron microscopy. The method was summarized as follows: S100A8 / A9 samples were dropped onto a glow-treated grid (Quantifoil Au300R1.2 / 1.3). Images were collected on a 300kV Thermo Fisher Krios G4 cryo-electron microscope equipped with a Falcon 4 camera. The magnification was 96,000x and the pixel size was 0.808. A total of 6,258 images were collected, with an electron dose of 10000 for each image. The focal length range is -1.0μm--2.0μm. The 6258 good micrographs collected were further processed using cryoSMART (Shuimu Biotechnology Co., Ltd.). The particles were automatically picked by Blob-picker, and then the best particles were selected as templates for Template picking after two rounds of 2D classification for 3D reconstruction, and the final resolution was Density map. AlphaFold2 prediction was performed based on the amino acid sequence, and the predicted results were matched with the calculated map using UCSF Chimera software, and then the amino acid side chains were manually adjusted using COOT software. Finally, the refined model was spatially refined using PHENIX software. The results showed (Figure 6) that the antigen core epitope was S100A8: D14, L21, G24, S100A9: E60, K72, D98, E99, P101, G102, H103. The core binding sites of the antibody are light chain: K52, heavy chain: S54, S57, Y59, K99, R101, P102, R104.

[0266] Example 4: In vitro biological activity assay of anti-S100A8 / A9 mAbs

[0267] 4.1 Inhibitory effect of anti-S100A8 / A9 mAbs on the release of inflammatory cytokines from hS100A8 / A9-activated human PBMCs

[0268] Anti-S100A8 / A9 monoclonal antibody was prepared at a starting concentration of 2000 μg / mL and serially diluted five-fold over eight steps. 50 μL was added to each well of a 96-well plate (purchased from Corning, Cat. No. 3599). hS100A8 / A9 protein was prepared at a concentration of 60 μg / mL and 50 μL was added to each well of the plate. The anti-S100A8 / A9 monoclonal antibody was mixed with the serial concentrations of the antibody. Both the antigen and antibody used in this experiment were pyrogenic to less than 2 EU / mL.

[0269] The ligand protein and the gradient diluted antibody were mixed evenly in equal volumes and incubated at room temperature for 30 min. Human PBMC cells were revived (purchased from Maishun Biotechnology Co., Ltd., catalog number PB010C) and the cell density was adjusted to 1.0×10 6 cells / mL, 100 μL per well was plated into the corresponding position of a 96-well plate and mixed evenly with the drug. A blank control group (PBMC only) and a positive control group (PBMC + S100A8 / A9) were also set up. The volume of medium in the control group was replenished, and the cell plate was placed in a 37°C CO2 incubator for approximately 20 hours before detection. All drug and antigen dilutions and cell density adjustments were made using RPMI 1640 complete medium (purchased from Thermo Fisher, catalog number A1049101) containing 10% FBS (purchased from Thermo Fisher, catalog number 10099141C). After the incubation period, the cell supernatant was diluted and assayed for cytokines using a human IL-6 ELISA kit (purchased from Beijing Dakoway Biotechnology Co., Ltd., catalog number 1110603) and a human TNF-α ELISA kit (purchased from Beijing Dakoway Biotechnology Co., Ltd., catalog number 1117203) according to the assay instructions. OD450 values ​​were measured and dose-effect curves were plotted. The results showed that the anti-S100A8 / A9 mAb described in this application inhibited the binding of hS100A8 / A9 to PBMC cell surface receptors and suppressed the release of the inflammatory factors IL-6 ( FIG. 7A ) and TNF-α ( FIG. 7B ).

[0270] 4.2 Anti-S100A8 / A9 mAbs inhibit the proliferation and toxicity of hS100A8 / A9 on mouse lymphoma EL-4 cells

[0271] hS100A8 / A9 protein was prepared at a concentration of 50 μg / mL, and 50 μL was added to each well of a 96-well plate (purchased from Corning, catalog number 3599). The EL-4 cell density was adjusted to 1.0×10 5cells / mL, 100 μL per well was added to a 96-well plate, mixed evenly with human S100A8 / A9 protein, and incubated for 6 h in a 37°C CO2 incubator. Anti-S100A8 / A9 monoclonal antibody was prepared at a starting concentration of 9240 μg / mL and diluted 3-fold in antibody diluent for a total of 8 steps. 50 μL was added to the corresponding positions in the 96-well plate at each well. All drug and antigen dilutions and cell density adjustments were performed using DMEM complete medium (purchased from Thermo Fisher, catalog number C11995500BT) supplemented with 10% horse serum (HS, purchased from EallBio, catalog number 03.16002A). A blank group (EL-4 only) and a negative control group (EL-4 + S100A8 / A9) were also set up. The volume of culture medium in the control group was supplemented, and the cell plates were placed in a 37°C CO2 incubator. After incubation for approximately 17 hours, CCK-8 reagent (purchased from EallBio, catalog number 03.17002DC) was added. After incubation for approximately 4 hours, the OD450 value was measured, and a dose-effect curve was plotted. The results show (Figure 8) that the hS100A8 / A9 antibody in this application can bind to hS100A8 / A9, block the proliferative toxicity of hS100A8 / A9 on mouse lymphocyte EL-4 cells, and promote the proliferation of EL-4 cells, showing a significant dose-effect relationship.

[0272] Example 5: Anti-S100A8 / A9 mAbs Block the Binding of hS100A8 / A9 to the Receptor

[0273] 5.1 Anti-S100A8 / A9 mAbs block the binding of hS100A8 / A9 to cell surface TLR4 or RAGE receptors

[0274] Anti-S100A8 / A9 monoclonal antibodies and isotype control antibodies (IgG) were prepared at a starting concentration of 800 μg / mL. Antibodies were serially diluted in antibody diluent in 10 or 8 3-fold (for TLR4) or 5-fold (for RAGE) steps, respectively. The dilutions were added to a 96-well plate (purchased from Corning, Cat. No. 3894) at a rate of 50 μL per well. Human S100A8 / A9 ligand protein was prepared at a concentration of 32 μg / mL (for TLR4) or 8 μg / mL (for RAGE), at a rate of 50 μL per well, and added to a 96-well plate. The ligand protein and serially diluted antibodies were mixed in equal volumes and incubated at room temperature for 30 minutes.

[0275] HEK293-TLR4 engineered cell lines were used to test the binding and blocking of hS100A8 / A9 to surface TLR4 receptors. HEK293-RAGE engineered cell lines were used to test the binding and blocking of hS100A8 / A9 to surface RAGE receptors. Single cell suspensions were obtained and counted. After centrifugation, the suspensions were resuspended in PBS buffer and the cell density was adjusted to 2.0×10 6 cells / mL, 100 μL per well was added to the corresponding position in the 96-well plate and incubated at 4°C for 1 hour. After incubation, the cells were washed once by centrifugation at 3000 rpm, the cell pellets were collected, 200 μL PBS buffer was added to each well, the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. FITC anti-His-Tag antibody prepared in advance (purchased from Biolegend, product number 362618) was added to the cell pellet, 100 μL / well, and incubated at 4°C for 1 hour. After taking out, 100 μL PBS buffer was added to each well and washed once at 3000 rpm. After resuspending in 200 μL PBS buffer, the cells were detected on a flow cytometer and the fluorescence signal in the FL1-A channel was collected. The results showed that the hS100A8 / A9 antibody of the present application can block the binding of hS100A8 / A9 to TLR4 (Figure 9A) or RAGE (Figure 9B) on the surface of the engineered cell line, showing a significant dose effect. No blocking effect was observed with the isotype control antibody.

[0276] 5.2 Anti-S100A8 / A9 mAbs Block the Binding of hS100A8 / A9 to PBMCs and THP-1 Cells

[0277] Anti-S100A8 / A9 monoclonal antibody was prepared at a starting concentration of 800 μg / mL. Eight 5-fold serial dilutions were performed in Antibody Diluent (for interaction with PBMCs) or 4-fold serial dilutions (for interaction with THP-1 cells). The dilutions were added to a 96-well plate (purchased from Corning, Cat. No. 3894) at a rate of 50 μL per well. Human S100A8 / A9 ligand protein was prepared at a concentration of 50 μg / mL and 50 μL per well was added to the 96-well plate. The ligand protein and serially diluted antibody were mixed in equal volumes and incubated at room temperature for 30 minutes.

[0278] PBMC cells and THP-1 cells were collected to obtain single cell suspension and counted. After centrifugation, they were resuspended in PBS buffer and the cell density was adjusted to 2.0×10 6cells / mL, 100 μL per well was added to the corresponding position in a 96-well plate and incubated at 4°C for 1 hour. After incubation, the cells were washed once by centrifugation at 3000 rpm, the cell pellets were collected, 200 μL PBS buffer was added to each well, the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. FITC anti-His-Tag antibody prepared in advance (purchased from Biolegend, item number 362618) was added to the cell pellet, 100 μL / well, and incubated at 4°C for 1 hour. After taking out, 100 μL PBS buffer was added to each well and washed once at 3000 rpm. After resuspending in 200 μL PBS buffer, the cells were detected on a flow cytometer and the fluorescence signal in the FL1-A channel was collected. The results showed that the anti-S100A8 / A9 monoclonal antibody of the present application can compete for the binding of hS100A8 / A9 to PBMC cells (Figure 10A), and can also compete for the binding of hS100A8 / A9 to THP-1 cells (Figure 10B). There was a significant dose effect.

[0279] Example 6: Activity of Genetically Engineered Mutants of Anti-S100A8 / A9 Monoclonal Antibodies

[0280] 6.1 Binding Activity of Single-Point Mutation Antibodies in the CDR Regions of Anti-S100A8 / A9 Monoclonal Antibodies

[0281] Alanine scanning and single-site-directed mutagenesis were performed on the six CDR regions to obtain single-point mutant antibodies. These single-point mutant antibodies were expressed and purified using the method described in 2.3 of Example 2, and the binding activity of the purified mutant antibodies was compared with that of the parent antibody using the ELISA method described in 2.4 of Example 2. After screening, it was found that the mutant antibodies shown in Table 3 below all retained the binding activity of the parent antibody (CDR sequences are shown in SEQ ID Nos: 1, 2, 3, 4, 5, and 6, respectively).

[0282] 6.2 Biological Activity of Anti-S100A8 / A9 Monoclonal Antibody CDR Single-Point Mutation Antibodies

[0283] The 38 single-point mutant antibodies listed in Table 3 below were expressed and purified using the method described in 2.3 of Example 2. The biological activity of the single-point mutant antibodies was assessed by inhibiting the release of IL-6 from human PBMCs activated by hS100A8 / A9. The specific assay method was as follows: Different mutants of the anti-S100A8 / A9 monoclonal antibody were prepared at a starting concentration of 800 μg / mL. Seven 5× dilutions of the antibody diluent were added to a 96-well plate (purchased from Corning, Cat. No. 3799), with 50 μL per well added. The hS100A8 / A9 ligand protein was prepared at a concentration of 80 μg / mL, with 50 μL per well added to the 96-well plate. The pyrogen content of both the antigen and antibody used in this experiment was less than 2 EU / mL. The ligand protein and the serially diluted antibody were mixed in equal volumes and incubated at room temperature for 30 minutes. Resuscitated human PBMC cells (purchased from Maishun Biotechnology Co., Ltd., catalog number PB010C) were adjusted to a cell density of 1.0 × 10 6 cells / mL, 100 μL per well was plated into the corresponding position of a 96-well plate and mixed evenly with the preincubation mixture. A blank control (PBMC only) and a positive control (PBMC + S100A8 / A9) were also set up. The culture medium volume of the control group was replenished, and the cell plate was placed in a 37°C CO2 incubator for approximately 20 h before detection. All antibody and antigen dilutions and cell density adjustments were made using RPMI 1640 complete medium (Thermo Fisher, Catalog No. A1049101) supplemented with 10% FBS (Thermo Fisher, Catalog No. 10099141C). After incubation, the cell supernatant was removed and diluted to the desired concentration. Cytokine detection was performed using a human IL-6 ELISA kit (Beijing Dakoway Biotechnology Co., Ltd., Catalog No. 1110603) according to the assay instructions. OD450 values ​​were measured, and dose-effect curves were plotted. The results showed ( FIG. 11 and Table 3 ) that the 38 mutants of the anti-S100A8 / A9 monoclonal antibody in the present application (as shown in Table 3 below) were able to inhibit the binding of hS100A8 / A9 to PBMC cell surface receptors and the release of IL-6, showing a significant dose-dependent effect.

[0284] The core HCDR sites are F29, Y32, A33, M34, Y59, A61, D62, Y100, S103, V105, F106, and S108. Mutations in these core amino acid sites resulted in approximately a 10-fold increase in cytological activity, including F29Y, F29W, Y59W, A61V, A61H, V105A, and F106Y. Mutations in these sites resulted in approximately a 2-fold increase in cytological activity, including F29H, A33V, Y100F, Y100H, and F106H. Antibodies obtained by mutation in other sites also retained good cytological activity, including M34L, A61L, S103H, F106W, Y32H, Y32F, M34I, Y59F, D62A, D62H, S103T, and S108A. The sequences of the HCDR regions of antibodies with good in vitro biological activity are shown in Table 3 below.

[0285] The core sites in the LCDR region are Y31, E49, D50, S51, K52, A92, and Q88. Mutations in these core amino acid sites resulted in approximately a 5-fold increase in cytological activity for A92I, approximately a 2-fold increase for Y31F and E49Q, and less than a 2-fold increase for K52H and A92L. Antibodies obtained by mutations in other sites also retained good cytological activity, including D50Q, D50H, K52P, Q88D, S51A, S51H, K52R, A92V, and S94A. The sequences of the LCDR regions of antibodies with excellent in vitro biological activity are shown in Table 3.

[0286] Table 3 Changes in antibody cytological activity after single-point mutation of CDR

[0287] 6.3 Binding Activity of Anti-S100A8 / A9 Monoclonal Antibody CDR Region Combination Mutations

[0288] Based on the results of Examples 6.1 and 6.2, combinatorial mutations were performed on multiple mutation sites in the CDR region to obtain mutant antibodies with combined mutations. The combined mutant antibodies were expressed and purified using the method described in 2.3 of Example 2, and the binding activity of the purified combined mutant antibodies was compared with that of the parent antibody using the ELISA method described in 2.4 of Example 2. After screening, it was found that compared with the parent antibody (CDR sequences are shown in SEQ ID Nos: 1, 2, 3, 4, 5, and 6, respectively), the combined mutant antibodies shown in Table 4 below all retained the binding activity of the parent antibody.

[0289] 6.4 Biological Activity of Anti-S100A8 / A9 Monoclonal Antibody CDR Combination Mutations

[0290] The 80 multi-point combination mutant antibodies listed in Table 4 below were expressed and purified using the method described in 2.3 of Example 2, and their biological activity against the multi-point combination mutant antibodies was tested using the method described in 6.2 of Example 6. The results ( FIG. 12 and Table 4 ) show that all 80 combination mutants of the anti-S100A8 / A9 monoclonal antibodies described in this application (as shown in Table 4 below) were able to inhibit the binding of hS100A8 / A9 to PBMC cell surface receptors and the release of IL-6, exhibiting a significant dose-dependent effect.

[0291] The core sites of the HCDR region are SEQ ID NOs: 27-64. The sequences of the HCDR regions of antibodies with good in vitro biological activity are shown in Table 4 below.

[0292] The core sites of the LCDR region are SEQ ID NOs: 65-109. The sequences of the LCDR regions of antibodies with good in vitro biological activity are shown in Table 4 below.

[0293] Table 4 Changes in antibody cytological activity after CDR combination mutations

[0294] Example 7: Pharmacokinetics of anti-S100A8 / A9 monoclonal antibodies

[0295] To investigate the in vivo metabolism of the anti-S100A8 / A9 monoclonal antibody described herein, five male C57BL / 6 mice were administered a 10 mg / kg dose via tail vein injection. Serum antibody concentrations were determined by blood sampling at various time points, with PK blood sampling occurring at 5 minutes, 2 hours, 4 hours, 8 hours, 1 day, 3 days, 7 days, and 14 days after administration.

[0296] Serum samples from the drug-drug collection site were assayed for total antibody and active antibody concentrations using ELISA. Goat anti-human Fc antibody (1:2000, Sigma-Aldrich, Product number: I8885) and antigen (1 μg / mL, hS100A8 / A9) were diluted in PBS, 100 μL / well, and coated overnight at 4°C. 3% skim milk powder (diluted in PBS) was added to the plate at 300 μL / well and blocked at 37°C for 2 hours. Antibody standards of known concentrations and mouse serum were added at 100 μL / well and incubated at 37°C for 1 hour. The plates were washed and horseradish enzyme-conjugated goat anti-human IgG (H+L) (1:5000, Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., Product number: ZB-2304) was added at 100 μL / well and incubated at 37°C for 45 minutes. The plate was washed again, TMB was added for color development for 10 min, and finally the stop solution was added. The absorbance at 450 nm was measured on a microplate reader. The standard curve was fitted and inserted into the standard curve to calculate the sample concentration. The pharmacokinetic constants of 5 mice were calculated using Phoenix WinNonlin.

[0297] The results showed ( FIG. 13 ) that the average half-life (t1 / 2) of the anti-S100A8 / A9 monoclonal antibody in the present application in 5 male C57BL / 6 mice was 6.6 days.

[0298] Example 8: Effects of anti-S100A8 / A9 monoclonal antibodies on cardiac damage and cardiac function in S100A8 / A9 humanized mice with acute myocardial infarction and myocardial ischemia / reperfusion injury

[0299] 8.1 Effects of different doses of anti-S100A8 / A9 monoclonal antibodies on cardiac injury and cardiac function in S100A8 / A9 humanized mice with acute myocardial infarction

[0300] To investigate the dose-dependent improvement of the anti-S100A8 / A9 monoclonal antibody of the present application on acute myocardial infarction in mice, 40 S100A8 / A9 humanized mice were selected and randomly divided into four groups: an IgG negative control group, a low-dose group, a medium-dose group, and a high-dose group of anti-S100A8 / A9 monoclonal antibody. Each group had 10 mice, half male and half female. The left coronary artery was ligated to establish an acute myocardial infarction model. Some mice died during the operation, and 8 mice were finally dosed in each group. Immediately after the myocardial infarction surgery, anti-S100A8 / A9 monoclonal antibody or IgG control was administered by intravenous injection into the medial canthus. The low, medium, and high doses of anti-S100A8 / A9 monoclonal antibody were 12 mg / kg, 16 mg / kg, and 24 mg / kg, respectively, and the IgG dose was 24 mg / kg. The mice were observed daily. 14 days after myocardial infarction, cardiac function of the mice was detected by echocardiography, and the left ventricular ejection fraction was calculated. The mice were sacrificed, and the heart tissues were obtained, fixed, paraffin-embedded, and sectioned. Masson's staining was performed, and the infarct area was measured and calculated.

[0301] The results showed that low, medium, and high doses of anti-S100A8 / A9 monoclonal antibodies could increase the left ventricular ejection fraction of mice after myocardial infarction compared with the IgG negative control group, and the difference was statistically significant (p<0.05). Compared with the low-dose group, the high-dose anti-S100A8 / A9 monoclonal antibody group had an increased left ventricular ejection fraction, and the difference was statistically significant (p<0.05) (Figure 14).

[0302] The low, medium, and high doses of anti-S100A8 / A9 monoclonal antibodies could reduce the myocardial infarction area compared with the IgG negative control group, and the difference was statistically significant (p<0.05). The high dose of anti-S100A8 / A9 monoclonal antibody group had a smaller myocardial infarction area than the low dose group, and the difference was statistically significant (p<0.05) (Figure 15).

[0303] 8.2 Effects of different administration times of anti-S100A8 / A9 monoclonal antibodies on cardiac injury and cardiac function in S100A8 / A9 humanized mice with acute myocardial infarction

[0304] To explore the administration time window for the anti-S100A8 / A9 monoclonal antibody of the present application to improve acute myocardial infarction in mice, 50 hS100A8 / A9 humanized mice were selected and randomly divided into 5 groups, 1 / 2 of each sex, including an IgG negative control group, an anti-S100A8 / A9 monoclonal antibody administration group 2 hours (h), 4 hours, 8 hours, and 10 hours after surgery. The left coronary artery was ligated to establish an acute myocardial infarction model. Some mice died during the myocardial infarction surgery, and the remaining 9 mice were administered to the IgG group, 7 mice each in the 2h and 4h groups, and 8 mice each in the 8h and 10h groups. The IgG negative control group was administered IgG control by intravenous injection into the medial canthus 2 hours after myocardial infarction surgery at a dose of 20 mg / kg. The other 4 groups were administered anti-S100A8 / A9 monoclonal antibody at a dose of 20 mg / kg 2, 4, 8, and 10 hours after surgery, respectively. Fourteen days after myocardial infarction, cardiac function was assessed by echocardiography, and left ventricular ejection fraction was calculated. Mice were sacrificed, and heart tissue was obtained, fixed, paraffin-embedded, sectioned, and stained with Masson's staining. Infarct size was then measured and calculated.

[0305] The results showed that anti-S100A8 / A9 monoclonal antibody administered 2, 4, and 8 hours after surgery could improve the left ventricular ejection fraction of mice after myocardial infarction compared with the IgG negative control group, and the difference was statistically significant (p<0.05); the group administered 10 hours after surgery did not improve the ejection fraction compared with the IgG negative control group, but the group administered 2 hours after surgery showed an improvement in the left ventricular ejection fraction compared with the group administered 10 hours after surgery, and the difference was statistically significant (p<0.05) (Figure 16).

[0306] Anti-S100A8 / A9 monoclonal antibody administered 2, 4, and 8 hours after surgery could reduce the myocardial infarction area of ​​mice compared with the IgG negative control group, and the difference was statistically significant (p<0.05); there was no significant improvement in the myocardial infarction area in the group 10 hours after surgery compared with the IgG negative control group; the myocardial infarction area in the groups administered 2 hours and 4 hours after surgery was reduced compared with the 8-hour group, and the difference was statistically significant (p<0.05); the myocardial infarction area in the groups administered 2 hours and 4 hours after surgery was reduced compared with the 10-hour group, and the difference was statistically significant (p<0.05) (Figure 17).

[0307] The results showed that the therapeutic time window for administration of anti-S100A8 / A9 monoclonal antibodies after myocardial infarction is within 8 hours.

[0308] 8.3 Effect of anti-S100A8 / A9 monoclonal antibodies on cardiac function after myocardial ischemia / reperfusion in S100A8 / A9 humanized mice

[0309] To investigate the effect of the anti-S100A8 / A9 mAb of this application on improving cardiac function after myocardial infarction / recanalization in mice, 20 humanized S100A8 / A9 mice were randomly divided into two groups: an IgG negative control group and an anti-S100A8 / A9 mAb treatment group. Each group consisted of 10 mice, half male and half female. The left coronary artery was ligated for 60 minutes, followed by ligation and recanalization, mimicking post-MI thrombolysis or PCI recanalization. During the procedure, some mice died or modeling failed, resulting in the remaining 7 mice in the IgG negative control group and 9 mice in the anti-S100A8 / A9 mAb group receiving the drug. Immediately after recanalization, anti-S100A8 / A9 mAb or IgG was administered at a dose of 20 mg / kg. Mice were observed daily postoperatively. Cardiac function was assessed by echocardiography 28 days after drug administration, and left ventricular ejection fraction was calculated.

[0310] The results showed that anti-S100A8 / A9 mAb could significantly increase the left ventricular ejection fraction of mice after cardiac ischemia / reperfusion compared with the IgG negative control group (p<0.05), and improve cardiac function ( FIG18 ).

[0311] It should be noted that the above are only preferred embodiments of the present application and are not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Although specific embodiments have been described, it is possible that the applicant or other persons skilled in the art may have or currently cannot foresee alternatives, modifications, changes, improvements, and substantial equivalents of the above embodiments. Therefore, the appended claims submitted and the claims that may be amended are intended to cover all such alternatives, modifications, changes, improvements, and substantial equivalents. It is important that as technology evolves, many of the elements described herein may be replaced by equivalent elements that appear after the present application.

Claims

1. A fully human antibody or antigen-binding fragment thereof, characterized in that: Its complementarity determining region (CDR) defined according to the KABAT system has the following amino acid sequence: The amino acid sequence of the heavy chain CDR1 includes GX1TX2SX3X4X5X6X7, wherein X1 is F or Y, X2 is F, H, Y or W, X3 is S or D, X4 is Y, H or F, X5 is A or V, X6 is M, I or L, and X7 is S or A; The amino acid sequence of the heavy chain CDR2 includes AISGX8GGSX9X 10 YX 11 X 12 SVKG, where X8 is S or H, X9 is T or H, X 10 is Y, W or F, X 11 A, H, V or L, X 12 is D, A or H; The amino acid sequence of the heavy chain CDR3 includes KX 13 RPX 14 RX 15 X 16 DX 17 , X 13 is Y, F or H, X 14 S, T or H, X 15 V or A, X 16 F, Y, H or W, X 17 is S, A, F, C, H or Y; The amino acid sequence of the light chain CDR1 includes SGDALX 18 DKX 19 X 20 X 21 , where X 18 G or H, X 19 Y or F, X 20 A or M, X 21 is S or A; The amino acid sequence of the light chain CDR2 includes X 22 X 23 X 24 X 25 RX 26 X 27 , where X 22 E or Q, X 23 D, H or Q, X 24 is S, A or H, X 25 K, H, R, P or N, X 26 is P, Q, D or N, X 27 is S, D, Y or W; as well as The amino acid sequence of the light chain CDR3 includes X 28 SNDX 29 DX 30 X 31 W, where X 28 For Q or D, X 29 A, V, I or L, X 30 S or A, X 31 V, Y, W or R.

2. The antibody or antigen-binding fragment thereof according to claim 1, wherein the complementarity determining region has the amino acid sequence shown below: The heavy chain CDR1 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 1, 23-34; The heavy chain CDR2 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 2 and 35-48; The heavy chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 3 and 49-64; The light chain CDR1 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 4, 65-69; The light chain CDR2 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 5, 70-95; and The light chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 6, 96-109.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the amino acid sequence of the complementarity determining region is selected from at least one of the following groups (1) to (119): (1) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 6, respectively; (2) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 24, 2, 3, 4, 5, and 6, respectively; (3) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 25, 2, 3, 4, 5, and 6, respectively; (4) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 26, 2, 3, 4, 5, and 6, respectively; (5) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 29, 2, 3, 4, 5, and 6, respectively; (6) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 30, 2, 3, 4, 5, and 6, respectively; (7) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 31, 2, 3, 4, 5, and 6, respectively; (8) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 32, 2, 3, 4, 5, and 6, respectively; (9) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 33, 2, 3, 4, 5, and 6, respectively; (10) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 37, 3, 4, 5, and 6, respectively; (11) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 38, 3, 4, 5, and 6, respectively; (12) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 39, 3, 4, 5, and 6, respectively; (13) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 40, 3, 4, 5, and 6, respectively; (14) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 41, 3, 4, 5, and 6, respectively; (15) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 42, 3, 4, 5, and 6, respectively; (16) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 43, 3, 4, 5, and 6, respectively; (17) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 49, 4, 5, and 6, respectively; (18) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 50, 4, 5, and 6, respectively; (19) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 51, 4, 5, and 6, respectively; (20) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 52, 4, 5, and 6, respectively; (21) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 53, 4, 5, and 6, respectively; (22) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 54, 4, 5, and 6, respectively; (23) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 55, 4, 5, and 6, respectively; (24) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 56, 4, 5, and 6, respectively; (25) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 57, 4, 5, and 6, respectively; (26) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 66, 5, and 6, respectively; (27) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 70, and 6, respectively; (28) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 71, and 6, respectively; (29) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 72, and 6, respectively; (30) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 73, and 6, respectively; (31) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 74, and 6, respectively; (32) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 75, and 6, respectively; (33) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 76, and 6, respectively; (34) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 77, and 6, respectively; (35) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 96, respectively; (36) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 5, and 97, respectively; (37) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 98, respectively; (38) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 3, 4, 5, and 99, respectively; or (39) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 4, 5, and 100, respectively. (40) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 38, 3, 4, 5, and 97, respectively. (41) The heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are the amino acid sequences shown in SEQ ID NOs: 1, 2, 57, 4, 5, and 100, respectively. (42) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 5, and 99, respectively; (43) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 73, and 6, respectively; (44) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 2, 57, 4, 73, and 100, respectively; (45) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 3, 4, 73, and 100, respectively; (46) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 53, 66, 5, and 6, respectively; (47) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 4, 5, and 99, respectively; (48) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 66, 5, and 99, respectively; (49) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 38, 57, 4, 73, and 99, respectively; (50) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 4, 73, and 100, respectively; (51) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 31, 57, 4, 76, and 100, respectively; (52) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 4, 73, and 99, respectively; (53) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 53, 4, 73, and 100, respectively; (54) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 2, 53, 66, 5, and 99, respectively; (55) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 39, 53, 66, 5, and 100, respectively; (56) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 73, and 104, respectively; (57) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 66, 73, and 104, respectively; (58) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 46, 57, 4, 73, and 104, respectively; (59) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 64, 4, 73, and 104, respectively; (60) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 63, 4, 73, and 104, respectively; (61) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 57, 4, 85, and 100, respectively; (62) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 31, 42, 57, 4, 87, and 100, respectively; (63) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 57, 4, 73, and 106, respectively; (64) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 63, 66, 73, and 100, respectively; (65) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 57, 4, 85, and 104, respectively; (66) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 25, 42, 63, 4, 73, and 104, respectively; (67) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 29, 42, 62, 4, 87, and 100, respectively; (68) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 32, 42, 63, 4, 73, and 105, respectively; (69) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 63, 4, 85, and 100, respectively; (70) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 47, 63, 66, 73, and 100, respectively; (71) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 63, 66, 73, and 104, respectively; (72) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 63, 66, 73, and 104, respectively; (73) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 25, 42, 63, 4, 85, and 104, respectively; (74) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 46, 64, 4, 87, and 104, respectively; (75) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 48, 63, 66, 85, and 100, respectively; (76) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 43, 63, 66, 73, and 106, respectively; (77) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 88, and 100, respectively; (78) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 89, and 100, respectively; (79) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 4, 90, and 100, respectively; (80) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 92, and 100, respectively; (81) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 42, 57, 4, 91, and 100, respectively; (82) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 94, and 100, respectively; (83) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 93, and 100, respectively; (84) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 42, 57, 4, 93, and 109, respectively; (85) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 23, 42, 57, 4, 73, and 100, respectively; (86) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 29, 42, 57, 4, 73, and 100, respectively; (87) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 57, 4, 73, and 108, respectively; (88) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 73, and 107, respectively; (89) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 73, and 109, respectively; (90) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 73, and 100, respectively; (91) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 69, 73, and 100, respectively; (92) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 73, and 109, respectively; (93) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 93, and 109, respectively; (94) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 65, 86, and 100, respectively; (95) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 86, and 100, respectively; (96) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 67, 73, and 108, respectively; (97) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 57, 4, 95, and 108, respectively; (98) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 57, 4, 94, and 108, respectively; (99) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 100, respectively; (100) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 73, and 100, respectively; (101) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 68, 73, and 100, respectively; (102) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 108, respectively; (103) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 107, respectively; (104) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 73, and 109, respectively; (105) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 69, 73, and 100, respectively; (106) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 73, and 109, respectively; (107) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 45, 57, 4, 86, and 100, respectively; (108) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 4, 93, and 109, respectively; (109) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 45, 57, 65, 86, and 100, respectively; (110) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 34, 42, 57, 65, 73, and 100, respectively; (111) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 44, 57, 68, 73, and 100, respectively; (112) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 1, 45, 57, 65, 73, and 100, respectively; (113) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 1, 45, 57, 69, 73, and 100, respectively; (114) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 1, 45, 57, 4, 86, and 100, respectively; (115) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 28, 42, 57, 4, 73, and 108, respectively; (116) heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 have the amino acid sequences shown in SEQ ID NOs: 27, 42, 59, 4, 73, and 108, respectively; (117) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are shown in SEQ ID NOs: 27, 42, 58, 4, 73, and 108, respectively; (118) the amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 60, 4, 73, and 108, respectively; (119) The amino acid sequences of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are SEQ ID NOs: 27, 42, 61, 4, 73, and 108, respectively.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein the heavy chain variable region has an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the sequence shown in SEQ ID NO: 7, and the light chain variable region has an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to the sequence shown in SEQ ID NO:

8. 5 . The antibody or antigen-binding fragment thereof according to claim 1 , further comprising a heavy chain constant region as shown in SEQ ID NO: 9 and a light chain constant region as shown in SEQ ID NO:

10.

6. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein the antibody or antigen-binding fragment thereof comprises a single-chain antibody, a single-domain antibody, a diabody, a triabody, a disulfide-stabilized antibody, a nanobody, a Fab fragment, a Fab' fragment, a (Fab')2 fragment, a minibody, a monoclonal antibody, a polyclonal antibody, a monospecific antibody, a bispecific antibody, a multispecific antibody, a monovalent antibody, a multivalent antibody, a chimeric antibody, a fusion protein comprising the antigen-binding site of an antibody, or a whole antibody immunoglobulin IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD.

7. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 6, wherein the heavy chain is an amino acid sequence obtained by mutation at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 positions in the sequence shown in SEQ ID NO: 11, wherein the mutation comprises an insertion, deletion and / or substitution.

8. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, wherein the mutation is a substitution, and the substituted amino acid position is selected from at least one of positions 27, 29, 31, 32, 33, 34, 35, 54, 58, 59, 61, 62, 100, 103, 105, 106, and 108 of the sequence shown in SEQ ID NO:

11.

9. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 8, wherein the light chain variable region is an amino acid sequence obtained by mutation at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 positions in the sequence shown in SEQ ID NO: 12, wherein the mutation comprises an insertion, deletion and / or substitution.

10. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, wherein the mutation is a substitution, and the substituted amino acid position is selected from at least one of positions 28, 31, 32, 33, 49, 50, 51, 52, 54, 55, 88, 92, 94, and 95 of the sequence shown in SEQ ID NO:

12.

11. A polynucleotide, characterized in that The polynucleotide encodes the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10.

12. A recombinant vector, characterized in that The recombinant vector comprises the polynucleotide according to claim 11.

13. A host cell, characterized in that The host cell comprises the polynucleotide according to claim 11 and / or the recombinant vector according to claim 12.

14. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10.

15. A kit, characterized in that The kit comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10.

16. A multispecific antibody, characterized in that It comprises the antibody or antigen-binding fragment according to any one of claims 1 to 10, and further comprises one or more second antibodies or antigen-binding fragments thereof that specifically bind to other antigens.

17. The multispecific antibody according to claim 16, wherein the second antibody or antigen-binding fragment thereof is selected from a full-length antibody, Fab, Fab", (Fab")2, Fv, scFv, scFv-scFv, miniantibody, diabody or sdAb.

18. An antibody conjugate, characterized in that The invention comprises the antibody or antigen-binding fragment according to any one of claims 1 to 10 and a second functional structure, wherein the second functional structure is selected from Fc, a radioisotope, a half-life extending moiety, a detectable label and a drug.

19. The antibody conjugate according to claim 18, wherein: The half-life-extending structural moiety is selected from the group consisting of an albumin binding structure, a transferrin binding structure, a polyethylene glycol molecule, a recombinant polyethylene glycol molecule, human serum albumin, a fragment of human serum albumin, and an albumin polypeptide that binds to human serum albumin; The detectable label is selected from the group consisting of a fluorophore, a chemiluminescent compound, a bioluminescent compound, an enzyme, an antibiotic resistance gene, and a contrast agent; and / or The drug is selected from the group consisting of cytotoxins and immunomodulators.

20. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, the multispecific antibody according to claim 16 or 17, and / or the antibody conjugate according to claim 18 or 19 in the preparation of a drug, reagent, or kit for detecting, preventing, alleviating, or treating a disease. The use according to claim 20, wherein the disease is associated with increased expression levels of oligomers, dimers, tetramers or multimers of tetramers or higher of S100A8 and S100A9.

22. The use according to claim 20, wherein the disease comprises myocardial infarction and myocardial ischemia / reperfusion injury.

23. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, the multispecific antibody according to claim 16 or 17, and / or the antibody conjugate according to claim 18 or 19 in the preparation of a drug, reagent, or kit for reducing myocardial infarction area, improving cardiac function, or improving microcirculatory function.

24. A method for detecting, preventing, alleviating or treating a disease, the method comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, the pharmaceutical composition according to claim 14, the kit according to claim 15, the multispecific antibody according to claim 16 or 17, and / or the antibody conjugate according to claim 18 or 19.

25. The method of claim 24, wherein the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers of tetramers or greater of S100A8 and S100A9.

26. The method of claim 24, wherein the disease comprises ischemic organ damage and organ ischemia / reperfusion injury associated with S100A8 / A9, S100A8, S100A9.

27. A method for reducing myocardial infarction size, improving cardiac function, or improving microcirculatory function, the method comprising administering to a subject in need thereof the antibody or antigen-binding fragment thereof of any one of claims 1-10, the pharmaceutical composition of claim 14, the kit of claim 15, the multispecific antibody of claim 16 or 17, and / or the antibody conjugate of claim 18 or 19.

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