Anti-s100a8 / a9 antibodies and uses thereof

Abstract

The invention provides an anti-S100A8 / A9 antibody and application of the anti-S100A8 / A9 antibody. The antibody or the antigen binding fragment and the mutant thereof provided by the invention can block the biological function of S100A8 / A9. The antibody or the antigen binding fragment provided by the invention is expected to be developed into a clinical drug, so that the cardiac infarction area of an AMI patient is reduced, the occurrence of heart failure is reduced, the AMI patient is benefited for the whole life, and the antibody or the antigen binding fragment has important clinical significance and development value.

Description

Technical Field

[0001] 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

[0002] Cardiovascular disease (CVD) is the leading cause of death and ranks first in the global disease burden. Acute myocardial infarction (AMI) is a severe type of CHD. When the coronary artery is acutely and persistently ischemic and hypoxic, it causes myocardial cell necrosis and is the main cause of death and disability. The current treatment method is to open the infarction-related artery as soon as possible to restore myocardial blood perfusion, and choose thrombolysis, percutaneous coronary intervention (PCI) or thrombolysis followed by PCI according to the patient's condition. The popularity of PCI surgery has improved the treatment of AMI, and reperfusion has effectively prevented further necrosis of myocardial cells. However, it still faces two problems: first, the average time for AMI patients to arrive at the hospital is 5.5 hours after the onset of the disease, so the blocked blood vessels are usually reopened 6 hours after the onset of myocardial infarction. During this period, myocardial cells have already undergone necrosis due to ischemia and hypoxia, but there is no early treatment drug that can reduce the area of ​​myocardial infarction; second, reperfusion injury (I / R) will also be induced after the blood vessels are reopened. In addition, part of the myocardium has already undergone necrosis before reperfusion. The occurrence of decreased cardiac function and even heart failure is still a prominent problem, and the current clinical drug application cannot improve this situation.

[0003] The heart damage caused by AMI is the result of the synergistic effects of multiple cells and pathological processes. For example, both AMI and I / R will cause the infiltration of a large number of inflammatory cells. In animal experiments, inhibiting inflammation can reduce post-MI damage and improve cardiac function. However, many clinical trials of anti-inflammatory treatments for patients with MI have ended in failure, suggesting that broad-spectrum inflammation inhibition is not advisable in the treatment of ischemic heart disease, and more specific and targeted targets need to be developed.

[0004] S100A8 and S100A9 (also known as MRP8 and MRP14) belong to the calcium-binding protein S100 family. Human S100A8 protein consists of 98 amino acids with a molecular weight of 10.8kD; S100A9 consists of 113 amino acids with a molecular weight of 13.2kD. The two can form heterodimers, heterotetramers, and oligomers, mainly in the form of dimers, and each molecule can also form homomers. Due to the poor stability of homodimers, they usually exert biological functions in the form of heterodimers and tetramers under pathological and physiological conditions. S100A8 / S100A9 is mainly expressed by neutrophils, accounting for about 40% of their cytoplasmic proteins. When inflammatory cells are stressed, S100A8 / S100A9 is released as an alarmin, acting on Toll-like receptor 4 (TLR4) and receptor for advanced glycation end products (RAGE) to regulate immune inflammatory responses.

[0005] 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 genes related to inflammatory response, and the increase in S100A8 and S100A9 was the most significant among the differentially expressed genes related to inflammation, reaching a peak 6 hours after I / R, then gradually decreasing, and basically returning to the baseline level in 7 days. Subsequent studies by multiple teams (Circulation, 2020, 141(13): 1080-1094; Eur Heart J, 2019, 40(32): 2713-2723) have 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 were significantly increased in AMI patients 1 day after PCI, and the increased S100A8 / A9 levels were associated with the incidence of major adverse cardiovascular events (MACE), and were independent predictors of the long-term MACE incidence in AMI patients (Circulation, 2019, 140(9): 751-764). In addition, we also demonstrated that S100A8 / A9 acts on myocardial cell TLR4, regulates mitochondrial dysfunction and mediates 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). All of the above 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.

[0006] However, there are still many problems to be solved in applying S100A8 / A9 as a target for clinical treatment: first, the homology between human and mouse S100A8 / A9 is not high, and the neutralizing antibodies used in previous studies for mice are difficult to act on humans, and there are no fully human or humanized monoclonal antibodies for the existing commercial S100A9 antibodies; second, S100A8 / A9 mainly exerts its cardiac damage effect in the form of heterodimers and heterotetramers, and the neutralizing antibodies targeting S100A9 in the past cannot specifically target S100A8 / A9 dimers and tetramers. Therefore, we need to develop neutralizing antibodies against human S100A8 / A9 and verify the therapeutic effect in vivo.

[0007] It should be noted that the methods described in this section are not necessarily methods that have been previously conceived or adopted. Unless otherwise indicated, it should not be assumed that any method described in this section is considered to be 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. Summary of the invention

[0008] 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 according to the KABAT system has the following amino acid sequence: the amino acid sequence of the heavy chain CDR1 includes GX 1 TX 2 SX 3 X 4 X 5 X 6 X 7 , where X 1 F or Y, X 2 F, H, Y or W, X 3 For S or D, X 4 is Y, H or F, X 5 A or V, X 6 M, I or L, X 7 S or A; the amino acid sequence of the heavy chain CDR2 includes AISGX 8 GGSX 9 X 10 YX 11 X 12 SVKG, where X 8 S or H, X 9 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 is 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, X19 For 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 is 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.

[0009] 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.

[0010] According to one embodiment of the present application, a recombinant vector is also provided, wherein the recombinant vector comprises the polynucleotide described in the present application.

[0011] 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.

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

[0013] According to one embodiment of the present application, a kit is also provided, wherein the kit comprises the antibody or antigen-binding fragment thereof described in the present application.

[0014] 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.

[0015] 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 part that extends the half-life, a detectable marker and a drug.

[0016] According to one embodiment of the present application, there is also provided use of the antibody or antigen-binding fragment thereof described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application in the preparation of drugs, reagents, and kits for detecting, alleviating, preventing or treating diseases.

[0017] According to one embodiment of the present application, there is also provided the use of the antibody or antigen-binding fragment thereof described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application in the preparation of drugs, reagents, and kits for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function.

[0018] According to one embodiment of the present application, a method for detecting, alleviating, preventing or treating a disease is also provided, the method comprising administering the antibody or antigen-binding fragment thereof described in the present application, the pharmaceutical composition described in the present application, the kit described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application to a subject in need thereof.

[0019] 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 the antibody or its antigen-binding fragment described in the present application, the pharmaceutical composition described in the present application, the kit described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application to a subject in need thereof.

[0020] The present application provides a fully human monoclonal antibody that can block the biological function of the S100A8 / A9 heterodimer. In the present application, the constructed S100A8 and S100A9 humanized mouse models are used to verify the in vivo therapeutic effect of the antibody on ischemic heart disease. For example, the application of the antibody in the acute phase of MI and I / R can inhibit the cardiac damage caused by the target molecule, reduce the area of ​​myocardial infarction and the degree of late fibrosis, and improve the cardiac function of experimental animals. Therefore, the antibody or antigen-binding fragment provided by the present application is expected to be developed into a clinical drug, in order to reduce the area of ​​myocardial infarction in AMI patients, reduce the occurrence of heart failure, and benefit AMI patients for life, which has important clinical significance and development value.

[0021] 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

[0022] The accompanying drawings exemplarily illustrate the embodiments and constitute a part of the specification, and together with the text description of the specification, are used to explain the exemplary implementation of the embodiments. The embodiments shown are for illustrative purposes only and do not limit the scope of the claims. In all drawings, the same reference numerals refer to similar but not necessarily identical elements.

[0023] Figure 1 This is a graph showing the binding activity of the four candidate monoclonal antibodies in Example 2 to human S100A8 / A9 (hS100A8 / A9) antigen.

[0024] Figure 2 This is a graph showing the binding specificity of the anti-S100A8 / A9 monoclonal antibody to the hS100A8 / A9 homologous family protein in Example 2.

[0025] Figure 3 This is a graph showing the binding specificity of the anti-S100A8 / A9 monoclonal antibody to an unrelated protein in Example 2.

[0026] Figure 4 This is a graph showing the results of species-specific detection of anti-S100A8 / A9 monoclonal antibodies in Example 2.

[0027] Figure 5A This is a graph showing the stability of the anti-S100A8 / A9 monoclonal antibody in PBS in Example 2; Figure 5B This is a graph showing the stability results of the anti-S100A8 / A9 monoclonal antibody in Example 2 in C57BL / 6 mouse serum (m-Serum).

[0028] Figure 6 This is a diagram showing the results of epitope analysis of the binding of anti-S100A8 / A9 monoclonal antibodies to hS100A8 / A9 in Example 3.

[0029] FIG. 7 is a graph showing the inhibitory effect of the anti-S100A8 / A9 monoclonal antibody in Example 4 on the release of inflammatory factors stimulated by hS100A8 / A9 in human PBMC. Fig. 7A This is a graph showing the inhibitory effect of the anti-S100A8 / A9 monoclonal antibody on the release of IL-6 from human PBMC stimulated by hS100A8 / A9 in Example 4. Figure 7B This is a graph showing the inhibitory effect of the anti-S100A8 / A9 monoclonal antibody in Example 4 on the release of TNFα from human PBMC stimulated by hS100A8 / A9.

[0030] Figure 8This is a graph showing the proliferative toxicity of the anti-S100A8 / A9 monoclonal antibody in Example 4 on mouse lymphoma cells EL-4.

[0031] FIG. 9 shows that the anti-S100A8 / A9 monoclonal antibody in Example 5 can block the binding of hS100A8 / A9 to TLR4 on the surface of the engineered cell line ( Fig. 9A ) or RAGE( Fig. 9B ) combined with the result graph.

[0032] FIG. 10 shows the blocking effect of anti-S100A8 / A9 monoclonal antibody on hS100A8 / A9 and PBMC cells in Example 5 ( Fig. 10A ), THP-1 cells ( Fig. 10B ) combined result diagram.

[0033] Fig.11 This is a graph showing the in vitro biological activity results of the genetically engineered single-point mutants of the anti-S100A8 / A9 monoclonal antibody in Example 6.2.

[0034] Figure 12 Fig. 12A and 12B ) is a graph showing the in vitro biological activity of the genetically engineered combination mutants of the anti-S100A8 / A9 monoclonal antibodies in Example 6.4.

[0035] Fig.13 This is a graph showing the metabolic status of the anti-S100A8 / A9 monoclonal antibody in mice in Example 7.

[0036] Fig.14 The results of Example 8 are shown in Figure 8 showing the effects of low, medium and high doses of anti-S100A8 / A9 monoclonal antibodies on left ventricular ejection fraction after myocardial infarction in S100A8 / A9 humanized mice.

[0037] Fig.15 The graph shows the effects of low, medium and high doses of anti-S100A8 / A9 monoclonal antibodies on the myocardial infarction area in S100A8 / A9 humanized mice after myocardial infarction in Example 8.

[0038] Fig.16 The graph shows 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.

[0039] Fig.17 The graph shows the effect of the anti-S100A8 / A9 monoclonal antibody administered 2, 4, 8, and 10 hours after surgery on the infarct size of myocardial infarction in S100A8 / A9 humanized mice in Example 8.

[0040] Fig.18This is a graph showing the effect of the anti-S100A8 / A9 monoclonal antibody on the left ventricular ejection fraction after myocardial ischemia / reperfusion in S100A8 / A9 humanized mice in Example 8. DETAILED DESCRIPTION

[0041] Unless otherwise indicated, all numbers used in this specification and claims to indicate content, concentration, ratio, mass, volume, time, temperature, thickness, technical effect, etc. should be understood as modified by the term "about" or "approximately" in any case. Therefore, unless otherwise indicated, the numerical parameters listed in the following specification and the attached 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 the present 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.

[0042] Although the numerical ranges and parameters setting forth the broad scope of the present disclosure are approximate, the numerical values ​​set forth in the specific embodiments are provided as accurately as possible. However, any numerical value will inherently contain certain errors, which are necessarily caused by the standard deviation found in its corresponding test measurements. Each numerical range given in this specification will include each narrower numerical range that falls within the broader numerical range, just as if these narrower numerical ranges were all clearly written herein.

[0043] Unless otherwise specified or contradictory to the context, the terms or expressions used herein should be read in conjunction with the overall content of this article and as understood by those of ordinary skill in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art.

[0044] 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.

[0045] As used herein, "nucleic acid" and "polynucleotide" are used interchangeably to refer to a polymeric form of nucleotides of any length, including deoxyribonucleotides, ribonucleotides, combinations thereof, and analogs thereof.

[0046] In this application, "polypeptide" and "peptide" are used interchangeably to refer to amino acid polymers of any length. Therefore, polypeptides, oligopeptides, proteins, antibodies and enzymes are all included in the definition of polypeptides.

[0047] As described in this application, 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.

[0048] As described in this application, sequence "identity" refers to the degree to which two (nucleotide or amino acid) sequences have the same residue at the same position in the alignment, and is usually expressed as a percentage. Preferably, the identity is determined over the entire length of the compared sequences. Therefore, two copies with exactly the same sequence have 100% identity. Those skilled in the art know that some algorithms can be used to determine sequence identity, 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 Clustal W.

[0049] The terms “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 the like as used herein 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.

[0050] "Antibody" as described herein refers to a specific immunoglobulin for an antigenic site. Antibodies can be manufactured according to methods known in the art. Antibodies may be in the form of 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 the antigen binding site, as long as the antibody exhibits the desired biological binding activity.

[0051] The term "multispecific" as used herein refers to an antigen-binding protein having multiple epitope specificities (i.e., being able to specifically bind to two, three or more different epitopes on a biological molecule or being able to specifically bind to epitopes on two, three or more different biological molecules). The term "bispecific" as used herein refers to an antigen-binding protein having two different antigen-binding specificities.

[0052] 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 Edition 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., "IMGTunique 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).

[0053] The boundary of given CDR or FR may be 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 based on the most common antibody region sequence length, wherein insertion (such as "30a") is provided by inserting letters and deletion occurs in some antibodies. These two schemes place some insertions and deletions (indel) in different positions, thereby producing different numbering. The Contact scheme is based on the analysis of complex crystal structures, and is similar to the Chothia numbering scheme in many respects. 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.

[0054] Therefore, unless otherwise specified, it should be understood that the "CDR" of a given antibody or its region (such as its variable region) covers CDRs defined by any of the above schemes or other known schemes. For example, in the case where a specific CDR (such as CDR3) is specified to contain 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 FRs 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 herein adopts the Kabat scheme.

[0055] The "humanized" antibody described in this application refers to an antibody 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 by corresponding residues from a non-human antibody (e.g., an antibody from which CDR residues are derived), 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.

[0056] "Gene" described in this application is the whole nucleotide sequence required to produce a polypeptide chain or functional RNA. Therefore, the expression of a gene 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.

[0057] 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.

[0058] "Foreign" or "heterologous" as used herein are used interchangeably and refer to a source different from the native (original) organism, such as an organism from another species. "Heterologous gene" or "foreign gene" as used herein refers to a gene that does not naturally occur in the host organism and is introduced into the host organism by gene transfer.

[0059] The "vector" described in this application 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. Among them, plasmids are the most common form of vectors, which refer to circular double-stranded DNA that can accept exogenous nucleic acid fragments and replicate in prokaryotic or eukaryotic cells.

[0060] The term "expression vector" and "recombinant vector" used herein are interchangeable and refer to a vector containing an exogenous gene and a regulatory element for expression in a designated host organism. Introducing an expression vector into a suitable host organism can enable it to express the inserted target gene.

[0061] "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 may be in the form of a plasmid retained in the host organism, or may be integrated into the host organism genome. Host organisms containing the transformed gene are referred to as "transgenic" or "recombinant" or "transformed" organisms or "engineered" organisms. Transformation of host organisms with expression vectors may be performed using conventional techniques well known to those skilled in the art. When the host is a prokaryotic organism, competent cells that can absorb DNA may be harvested after the exponential growth phase and treated with CaCl 2 The method is well known in the art. If necessary, 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 here.

[0062] "Relieve" and "treat" and their synonyms as used herein 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 physically (e.g., stabilizing recognizable symptoms), physiologically (e.g., stabilizing physical parameters), or both, inhibiting the development of a disease, disorder and / or condition. "Relieve" and "treat" can also be slowing down the development or reversal of a disease, disorder and / or condition.

[0063] 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.

[0064] 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.

[0065] Antibodies or antigen-binding fragments

[0066] According to one embodiment of the present application, a fully human antibody or an antigen-binding fragment thereof is provided, characterized in that the 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

[0067] GX 1 TX 2 SX 3 X 4 X 5 X 6 X 7 , where X 1 F or Y, X 2 F, H, Y or W, X 3 For S or D, X 4 is Y, H or F, X 5 A or V, X 6 M, I or L, X 7 S or A; the amino acid sequence of the heavy chain CDR2 includes AISGX 8 GGSX 9 X 10 YX 11 X 12 SVKG, where X 8 S or H, X 9 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 13RPX 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 is 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

[0068] SGDALX 18 DKX 19 X 20 X 21 , where X 18 G or H, X 19 For 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 is 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.

[0069] In some embodiments, the complementary 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.

[0070] In some embodiments, the complementary determining region of the antibody or its antigen-binding fragment has an amino acid sequence as follows: 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, 35-48; the heavy chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs:3, 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.

[0071] 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):

[0072] (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 SEQ ID NOs: 1, 2, 3, 4, 5, and 6, respectively;

[0073] (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 SEQ ID NOs: 24, 2, 3, 4, 5, and 6, respectively;

[0074] (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 SEQ ID NOs: 25, 2, 3, 4, 5, and 6, respectively;

[0075] (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 SEQ ID NOs: 26, 2, 3, 4, 5, and 6, respectively;

[0076] (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;

[0077] (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;

[0078] (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 SEQ ID NOs: 31, 2, 3, 4, 5, and 6, respectively;

[0079] (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;

[0080] (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;

[0081] (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;

[0082] (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;

[0083] (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;

[0084] (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;

[0085] (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 SEQ ID NOs: 1, 41, 3, 4, 5, and 6, respectively;

[0086] (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;

[0087] (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;

[0088] (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;

[0089] (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;

[0090] (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;

[0091] (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 SEQ ID NOs: 1, 2, 52, 4, 5, and 6, respectively;

[0092] (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 SEQ ID NOs: 1, 2, 53, 4, 5, and 6, respectively;

[0093] (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 SEQ ID NOs: 1, 2, 54, 4, 5, and 6, respectively;

[0094] (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 SEQ ID NOs: 1, 2, 55, 4, 5, and 6, respectively;

[0095] (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 SEQ ID NOs: 1, 2, 56, 4, 5, and 6, respectively;

[0096] (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;

[0097] (26) 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, 66, 5, and 6, respectively;

[0098] (27) the 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, 70, and 6, respectively;

[0099] (28) the 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;

[0100] (29) the 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;

[0101] (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 SEQ ID NOs: 1, 2, 3, 4, 73, and 6, respectively;

[0102] (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 SEQ ID NOs: 1, 2, 3, 4, 74, and 6, respectively;

[0103] (32) 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, 75, and 6, respectively;

[0104] (33) 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, 76, and 6, respectively;

[0105] (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 SEQ ID NOs: 1, 2, 3, 4, 77, and 6, respectively;

[0106] (35) the 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 96, respectively;

[0107] (36) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 5, and 97, respectively;

[0108] (37) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 5, and 98, respectively;

[0109] (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

[0110] (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.

[0111] (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.

[0112] (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.

[0113] (42) the 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, 5, and 99, respectively;

[0114] (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 SEQ ID NOs: 1, 42, 57, 4, 73, and 6, respectively;

[0115] (44) the 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, 57, 4, 73, and 100, respectively;

[0116] (45) 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, 73, and 100, respectively;

[0117] (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 SEQ ID NOs: 1, 42, 53, 66, 5, and 6, respectively;

[0118] (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;

[0119] (48) 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, 66, 5, and 99, respectively;

[0120] (49) the 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;

[0121] (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 SEQ ID NOs: 1, 42, 57, 4, 73, and 100, respectively;

[0122] (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;

[0123] (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;

[0124] (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 SEQ ID NOs: 1, 42, 53, 4, 73, and 100, respectively;

[0125] (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;

[0126] (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 SEQ ID NOs: 1, 39, 53, 66, 5, and 100, respectively;

[0127] (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 SEQ ID NOs: 1, 42, 57, 4, 73, and 104, respectively;

[0128] (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 SEQ ID NOs: 1, 42, 57, 66, 73, and 104, respectively;

[0129] (58) 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, 46, 57, 4, 73, and 104, respectively;

[0130] (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 SEQ ID NOs: 1, 42, 64, 4, 73, and 104, respectively;

[0131] (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 SEQ ID NOs: 1, 42, 63, 4, 73, and 104, respectively;

[0132] (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;

[0133] (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;

[0134] (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 SEQ ID NOs: 1, 46, 57, 4, 73, and 106, respectively;

[0135] (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 SEQ ID NOs: 1, 42, 63, 66, 73, and 100, respectively;

[0136] (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;

[0137] (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 SEQ ID NOs: 25, 42, 63, 4, 73, and 104, respectively;

[0138] (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 SEQ ID NOs: 29, 42, 62, 4, 87, and 100, respectively;

[0139] (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 SEQ ID NOs: 32, 42, 63, 4, 73, and 105, respectively;

[0140] (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 SEQ ID NOs: 1, 46, 63, 4, 85, and 100, respectively;

[0141] (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;

[0142] (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 SEQ ID NOs: 1, 42, 63, 66, 73, and 104, respectively;

[0143] (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;

[0144] (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;

[0145] (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 SEQ ID NOs: 1, 46, 64, 4, 87, and 104, respectively;

[0146] (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 SEQ ID NOs: 1, 48, 63, 66, 85, and 100, respectively;

[0147] (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 SEQ ID NOs: 1, 43, 63, 66, 73, and 106, respectively;

[0148] (77) the 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, 88, and 100, respectively;

[0149] (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 SEQ ID NOs: 1, 42, 57, 4, 89, and 100, respectively;

[0150] (79) 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, 42, 57, 4, 90, and 100, respectively;

[0151] (80) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are represented by the amino acid sequences of SEQ ID NOs: 1, 42, 57, 4, 92, and 100, respectively;

[0152] (81) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are amino acid sequences represented by SEQ ID NOs: 1, 42, 57, 4, 91, and 100, respectively;

[0153] (82) the 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, 94, and 100, respectively;

[0154] (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 SEQ ID NOs: 1, 42, 57, 4, 93, and 100, respectively;

[0155] (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 SEQ ID NOs: 1, 42, 57, 4, 93, and 109, respectively;

[0156] (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;

[0157] (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 SEQ ID NOs: 29, 42, 57, 4, 73, and 100, respectively;

[0158] (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 SEQ ID NOs: 27, 42, 57, 4, 73, and 108, respectively;

[0159] (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;

[0160] (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;

[0161] (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;

[0162] (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;

[0163] (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;

[0164] (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;

[0165] (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;

[0166] (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;

[0167] (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;

[0168] (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;

[0169] (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 SEQ ID NOs: 27, 42, 57, 4, 94, and 108, respectively;

[0170] (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 SEQ ID NOs: 27, 45, 57, 4, 73, and 100, respectively;

[0171] (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;

[0172] (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;

[0173] (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 SEQ ID NOs: 27, 45, 57, 4, 73, and 108, respectively;

[0174] (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 SEQ ID NOs: 27, 45, 57, 4, 73, and 107, respectively;

[0175] (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 SEQ ID NOs: 27, 45, 57, 4, 73, and 109, respectively;

[0176] (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;

[0177] (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;

[0178] (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 SEQ ID NOs: 27, 45, 57, 4, 86, and 100, respectively;

[0179] (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;

[0180] (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;

[0181] (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;

[0182] (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;

[0183] (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;

[0184] (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 SEQ ID NOs: 1, 45, 57, 69, 73, and 100, respectively;

[0185] (114) 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, 4, 86, and 100, respectively;

[0186] (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;

[0187] (116) 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, 59, 4, 73, and 108, respectively;

[0188] (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 SEQ ID NOs: 27, 42, 58, 4, 73, and 108, respectively;

[0189] (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;

[0190] (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;

[0191] 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 applied to the present application. In some embodiments, the FR region of the heavy chain comprises the FR region in the heavy chain variable region domain shown in SEQ ID NO: 7. In some embodiments, the FR region of the light chain comprises the FR region in the light chain variable region domain shown in SEQ ID NO: 8.

[0192] 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%, 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%, 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.

[0193] 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.

[0194] 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.

[0195] 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 shown in SEQ ID NO:9, and the amino acid sequence of the light chain constant region is shown in SEQ ID NO:10.

[0196] In some embodiments, the antibody or antigen-binding fragment thereof may further optionally comprise a signal peptide. When the antibody or antigen-binding fragment thereof is expressed and purified using a prokaryotic expression system, suitable signal peptide sequences are known in the art, and any suitable signal peptide sequence may be applied to 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 a sequence as shown in SEQ ID NO: 15, and the light chain comprising the signal peptide has a sequence as shown in SEQ ID NO: 16.

[0197] 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 the light chain may not contain a signal peptide. In some preferred embodiments, the heavy chain not containing a signal peptide has a sequence as shown in SEQ ID NO: 11, and the light chain not containing a signal peptide has a sequence as shown in SEQ ID NO: 12.

[0198] In some embodiments, the antibody or its antigen-binding fragment includes but is not limited to a single-chain antibody, a single-domain antibody, a double-chain antibody, a three-chain antibody, 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 antibody's antigen-binding site, or a whole antibody immunoglobulin IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD.

[0199] Polynucleotide, recombinant vector, host cell

[0200] 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.

[0201] According to one embodiment of the present application, a recombinant vector is also provided, the recombinant vector comprising the polynucleotide described in the present application. Recombinant vectors suitable for expressing the antibody or antigen-binding fragment described in the present application are well known in the art and are not limited here.

[0202] In some embodiments, the recombinant vector further comprises a promoter. The promoter may be any suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by a host cell expressing the nucleic acid sequence. The promoter sequence contains a transcriptional regulatory sequence that mediates the expression of the antibody or antigen-binding fragment thereof. The promoter may be any nucleic acid sequence that is transcriptionally active in the selected host cell, including mutant, truncated, and hybrid promoters, and may be derived from a gene encoding an extracellular or intracellular protein or polypeptide that is homologous or heterologous to the host cell.

[0203] The recombinant vector of the present application can be constructed by methods 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.

[0204] According to one embodiment of the present application, a host cell is also provided, the host cell comprising the polynucleotide described in the present application and / or the recombinant vector described in the present application. The host cell can be selected according to the type of the expression vector. The polynucleotide and / or the expression vector can be delivered into the host cell using any suitable method known in the art, which is not limited here.

[0205] Pharmaceutical compositions, kits

[0206] According to one embodiment of the present application, a pharmaceutical composition is also provided, characterized in that the pharmaceutical composition comprises the antibody or antigen-binding fragment thereof described in the present application.

[0207] 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 by the present application to a mammal (eg, a human).

[0208] The term "pharmaceutically acceptable carrier" refers to a carrier, diluent or adjuvant used for the preparation or administration of the polypeptide, polynucleotide or recombinant vector provided in the present application, which is not an essential active ingredient and has no excessive toxicity after administration. Suitable pharmaceutically acceptable carriers are well known to those of ordinary skill in the art.

[0209] Among them, "physiologically acceptable carrier" refers to a carrier or diluent or adjuvant that does not cause significant stimulation to the organism and does not eliminate the pharmaceutical activity and properties of the polypeptide, polynucleotide or recombinant vector provided by the application. Suitable physiologically acceptable carriers are also well known to those of ordinary skill in the art.

[0210] 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.

[0211] 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.

[0212] 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.

[0213] Multispecific antibodies, antibody conjugates

[0214] 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.

[0215] 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, miniantibody, diabody or sdAb.

[0216] 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 by the present application with the second antibody or antigen-binding fragment thereof to form a multispecific antibody.

[0217] 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 part that extends the half-life, a detectable marker and a drug.

[0218] Suitable Fc, radioactive isotopes, structural parts that extend half-life, detectable markers 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 in the present application to the second functional structure using methods known in the art.

[0219] 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 the immunoglobulin heavy chain, which includes native Fc and variant Fc. "Native Fc" refers to a molecule or sequence containing a non-antigen binding fragment produced by digesting a whole antibody, whether in monomeric or multimeric form. The immunoglobulin source for producing native Fc is preferably derived from humans. The native Fc fragment is composed of monomeric polypeptides that can be connected in the form of dimers or multimers by covalent linkage (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), there are 1-4 intermolecular disulfide bonds between the monomer subunits of the native Fc molecule. 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 monomers, dimers and multimer forms. "Variant Fc" refers to an amino acid sequence that is different 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 a 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.

[0220] 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, cammycin, 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.

[0221] In some embodiments, the antibody conjugate comprises the anti-S100A8 / A9 antibody provided in the present application and a radioactive isotope to form a radionuclide drug conjugate (RDC). Examples of radioactive isotopes that can be used in the present application include, but are not limited to, At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, 99mTc, 123I, 18F, and 68Ga.

[0222] In some embodiments, the antibody conjugate comprises the anti-S100A8 / A9 antibody provided herein and a half-life extending structural portion to improve the half-life of the anti-S100A8 / A9 antibody. The half-life extending structural portion is selected from the group consisting of albumin binding structure, transferrin binding structure, polyethylene glycol molecule, recombinant polyethylene glycol molecule, human serum albumin, human serum albumin fragments and albumin polypeptides binding to human serum albumin.

[0223] In some embodiments, the antibody conjugate comprises an anti-S100A8 / A9 antibody provided herein and a detectable marker, wherein the detectable marker is selected from a fluorophore, a chemiluminescent compound, a bioluminescent compound, an enzyme, an antibiotic resistance gene, and a contrast agent. The term "detectable marker" in the present application means 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 according to the detection system used.

[0224] use

[0225] According to one embodiment of the present application, there is also provided use of the antibody or antigen-binding fragment thereof described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application in the preparation of drugs, reagents, and kits for detecting, preventing or treating diseases.

[0226] In some embodiments, the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers greater than tetramers 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 above tetramers, 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 (AdvSci (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 cardio-cerebral diseases (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.

[0227] According to one embodiment of the present application, there is also provided the use of the antibody or antigen-binding fragment thereof described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application in the preparation of drugs, reagents, and kits for reducing myocardial infarction area, improving cardiac function, or improving microcirculation function.

[0228] In the purposes described in the present application, the dosage of the antibody or antigen-binding fragment provided by the present application can depend on several factors, including the severity and responsiveness of the symptoms, the route of administration, the treatment time (several days to several months to several years), and the time to improve the symptoms. Those skilled in the art can adjust the dosage regimen to provide a therapeutic response according to the specific circumstances of the patient. For example, a single administration can be performed, several separate doses can be administered within a predetermined time period, or the dosage can 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 can also vary with the type and severity of the condition to be alleviated. For any specific subject, the specific dosage regimen can be adjusted over time according to individual needs and the professional judgment of the treating clinician.

[0229] Disease detection, prevention or treatment methods

[0230] According to one embodiment of the present application, a method for detecting, preventing or treating a disease is also provided, the method comprising administering the antibody or antigen-binding fragment thereof described in the present application, the pharmaceutical composition described in the present application, the kit described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application to a subject in need thereof.

[0231] In some embodiments, the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers greater than tetramers 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 above tetramers, 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 (AdvSci (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 cardio-cerebral 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, and S100A9.

[0232] 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 the antibody or its antigen-binding fragment described in the present application, the pharmaceutical composition described in the present application, the kit described in the present application, the multispecific antibody described in the present application, and / or the antibody conjugate described in the present application to a subject in need thereof.

[0233] 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, the route of administration, the treatment time (several days to several months to several years), and the time to improve the symptoms. Those skilled in the art may adjust the dosage regimen to provide a therapeutic response according to the specific circumstances of the patient. 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.

[0234] 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 as part of the disclosure of this application.

[0235] The exemplary embodiments of the present application will be described below 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 by no means intended to limit the scope of protection of the present application. The scope of protection of the present application is limited only by the claims. Therefore, it should be recognized by those of ordinary skill in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope of the present application. Similarly, for clarity and conciseness, the description of well-known functions and structures is omitted in the following description.

[0236] Example

[0237] If no specific techniques or conditions are specified in this embodiment, the techniques or conditions described in the literature in this field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be obtained through commercial purchase.

[0238] Example 1: Preparation of prokaryotic recombinant antigens from different species

[0239] S100A8 (Sino Biological, Catalog No. HG11138-CH, Uniprot: P05109, its amino acid sequence is shown in SEQ ID No: 110, its encoding nucleic acid sequence is shown in SEQ ID No: 111) and S100A9 (Sino Biological, Catalog No. HG11145-CH, Uniprot: P06702, its amino acid sequence is shown in SEQ ID No: 112, its encoding nucleic acid sequence is 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) cut with NdeI and BamHI to construct pET21-S100A8 or pET21-S100A9 vectors, and the co-expression experiment used the co-expression vector pET21-S100A8-S100A9. A similar method was used to construct and prepare recombinant proteins of different species, including human S100A8 / A9 (hS100A8 / A9), mouse S100A8 / A9 (mS100A8 / A9) and monkey S100A8 / A9 (cS100A8 / A9). A His-tag was designed at the C-terminus of the recombinant protein for purification. E. coli BL21 (DE3) T7 expression cells were used to transform the expression plasmid. The colony was inoculated in 20 mL LB containing 100 μg / mL ampicillin and cultured in a shake flask at 37°C for 2 h. 20 mL of the preculture was inoculated in 800 mL LB containing 100 μg / mL ampicillin. When the cell density reached OD600 of 0.5, 0.5 mM isopropyl β-d-1-thiogalactopyranoside (IPTG) was added and induced for 3 h. The E. coli was harvested by centrifugation and the centrifuged cells were washed with 0.15 M NaCl. Resuspend the cells in 80 mL of 50 mM Tris-HCl buffer (pH 7.5) containing 50 mM NaCl and 5 mM MgSO 4 , and the bacteria were broken by ultrasound on ice. The supernatant collected by centrifugation after ultrasound was purified by metal chelate affinity chromatography (Ni-NTA) to obtain a fusion recombinant protein with a His-tag tag. The target protein collected after purification was dialyzed and the solution of the target protein was replaced with PBS, and then aliquoted and stored at -80°C. The recombinant protein was depyrogenated according to the needs of different experiments, and then the pyrogen-free recombinant protein was aliquoted and stored at -80°C.

[0240] Example 2: Preparation and activity detection of anti-S100A8 / A9 antibodies

[0241] 2.1 Screening of anti-hS100A8 / A9 antibodies using phage antibody library

[0242] Immunotubes were coated with hS100A8 / A9-His (hereinafter referred to as hS100A8 / A9) antigen prepared in Example 1, 20 μg / mL, 1 mL / tube, overnight at 4°C; the next day, PBS was washed 4 times, 3 minutes / time, and then blocked with 1.5 mL of 2% BSA at 37°C for 2 hours. The phage library displaying single-chain antibody (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, with the input amount of 10 9 -10 12 / tube, combined at 37℃ for 1h. After washing with PBST and PBS, 1mL0.2mol / L glycine-hydrochloric acid (pH2.2) was added for elution, and oscillated at room temperature for 10min. Immediately after aspiration, it was neutralized with 1mol / LTris of pH 8.8 to about pH7.0. The neutralized phage was infected with XL1-Blue activated bacteria in the logarithmic growth phase, and the cells were placed in a 37℃ incubator for 20-30min, and then placed in a shaker for 30min / 37℃ / 150rpm shaking culture; then 1% and 0.1% of the bacterial solution were plated for counting, and the remaining bacterial solution was centrifuged at 4000rpm for 10min, the supernatant was discarded, and the bacteria were spread on a 2YTCG solid plate, and then cultured overnight at 37℃. The next day, the bacteria on the plate were collected and inoculated into 2YTCG liquid medium with shaking until the logarithmic growth phase, then infected with M13KO7, left at room temperature for 15-30 minutes, then cultured at 37°C / 150rpm for 1 hour, and then 50μg / mL kanamycin was added and cultured at 30°C overnight. The next day, PEG8000 / NaCl was used to precipitate and purify the phage, which can be used for the next round of screening. A total of three rounds of phage library enrichment screening were performed.

[0243] 2.2 Identification of monoclonal anti-hS100A8 / A9 single-chain antibody

[0244] A single colony obtained after three rounds of phage screening was selected and inoculated into 200 μL of 2YTCTG medium. The culture was continued at 37°C and 220 rpm until the logarithmic growth phase. Then 10 8The helper phage M13KO7 was infected at room temperature for 20 minutes, then cultured at 37°C, 150rpm for 1 hour, and then an equal volume of 2×YTCTKI (kanamycin 50μg / mL, IPTG 0.2mmol / L) was added, and cultured at 200rpm and 30°C overnight. The next day, the supernatant was collected by centrifugation, BSA was added to a final concentration of 2%, Tween-20 was added to a final concentration of 0.1%, and it was incubated at 37°C for 15 minutes for ELISA identification. The hS100A8 / A9 antigen was coated on a 96-well ELISA plate, 200ng / well, overnight at 4°C. The next day, the ELISA plate was blocked with PBST-4% milk, 250μL / well, 37°C, 2h. The blocking solution was discarded, and the blocked phage antibody was added, and it was incubated at 37°C for 1h. The liquid was discarded and washed 3 times with PBST. Anti-M13 Antibody (HRP) (Sino Biological, Cat: 11973-MM05T-H) was diluted with blocking solution and added to the ELISA plate, and then incubated at 37°C for 30 minutes. Washed with PBST three times, OPD substrate colorimetric solution was added, and the color was incubated at room temperature for about 10 minutes, and then stained with 2M H 2 SO 4 Stop color development and measure the light absorption value with an enzyme-labeled instrument.

[0245] 2.3 Conversion of single-chain antibodies to whole antibodies and expression and purification

[0246] The anti-hS100A8 / A9 single-chain antibody obtained after screening was used to prepare the full-length antibody. Primers LF (the corresponding nucleotide sequence is shown in SEQ ID No: 19) and LR (the corresponding nucleotide sequence is shown in SEQ ID No: 20) were used to amplify the VL variable region gene; primers HF (the corresponding nucleotide sequence is shown in SEQ ID No: 21) and HR (the corresponding nucleotide sequence is 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 respectively by homologous recombination method, and after transforming Escherichia coli Top10, single clones were picked to sequence and identify the recombinant plasmids, and the correct heavy chain and light chain expression vectors of the fully human antibody were selected. 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 whole antibody. The expression supernatant was purified by Protein A affinity chromatography, and the purified whole 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.

[0247] 2.4 Binding activity detection of anti-S100A8 / A9 mAbs

[0248] The binding activity of the four candidate monoclonal antibodies to the hS100A8 / A9 antigen was analyzed by ELISA. The monoclonal antibodies expressed and purified were coated on 96-well plates (0.5 μg / mL, 50 μL / well, coated overnight at 4°C), and the affinity of the hS100A8 / A9 antigen at different dilutions to the monoclonal antibodies was detected. The starting concentration of the hS100A8 / A9 antigen was 100 μg / mL, and 5-fold or 3-fold gradient dilutions were performed to set 8 dilutions. The HRP-conjugated secondary antibody used was HRP-conjugated 6×His, His-Tag Monoclonal antibody (proteintech, Cat No: HRP-66005) (results see Figure 1 ), the sequence related information is shown in Table 1 below.

[0249] Table 1 Antibody related sequences

[0250]

[0251]

[0252]

[0253] Note: In the amino acid sequences of the heavy chain / light chain above, the part marked with double underlined lines at the downstream of the sequence is the constant region sequence; the part marked with bold font in the middle of the sequence is the complementarity-determining region (CDR) sequence.

[0254] 2.5 Activity analysis of anti-S100A8 / A9 mAbs

[0255] 2.5.1 Reichert 4SPR detection of anti-S100A8 / A9 monoclonal antibody affinity experiment

[0256] Anti-S100A8 / A9 monoclonal antibodies were covalently coupled to the SR7000 GOLD SENSOR SLIDE biosensor chip (Reichert, PART NO: 13206066). When the target molecules flowed through the chip surface, they could be captured by the anti-S100A8 / A9 monoclonal antibodies on the chip. The reaction signals were detected in real time using a Reichert 4SPR instrument to obtain the antigen-antibody binding and dissociation curves, and then the affinity value was obtained by fitting. The present application further used Reichert 4SPR to detect the affinity of the screened anti-S100A8 / A9 monoclonal antibodies to the human heterodimeric antigen hS100A8 / A9 and the monkey heterodimeric antigen cS100A8 / A9, respectively. 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 the affinity to cS100A8 / A9 is weaker than that to hS100A8 / A9.

[0257] Table 2 Affinity test results

[0258] Antibody antigen KD(M) Anti-S100A8 / A9 monoclonal antibody hS100A8 / A9 (8.95E±0.54)-09 Anti-S100A8 / A9 monoclonal antibody cS100A8 / A9 (2.28E±0.13)-08

[0259] 2.5.2 Specificity of anti-S100A8 / A9 mAbs and proteins in the same family

[0260] 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: P06704) were coated on 96-well plates. niprot: P31151; S100A8, Uniprot: P05109; S100A9, Uniprot: P06702; S100A10, Uniprot: P6090 3; S100A11, Uniprot: P31949; S100A12, Uniprot: P80511; S100A13, Uniprot: Q99584; S100A14, Uniprot: Q9HCY8; S100A15, Uniprot: Q86SG5; S100A16, Uniprot: Q96FQ6; S100A7L2, Uniprot: Q 5SY68; S100B, Uniprot: P04271; S100G, Uniprot: P29377; S100P, Uniprot: P25815; S100Z, Unip rot: Q8WXG8; S100A8 / A9) (0.5μg / mL, 50μL / well, coated overnight at 4℃), the next day, the liquid in the plate was discarded, and the plate was blocked with blocking solution (4% milk-PBS) at 37℃ for 1 hour, and anti-S100A8 / A9 monoclonal antibody was added, with a concentration of 20μg / mL, and bound to the plate at 37℃ for 1h, then washed 4 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 this application specifically bound to hS100A8 / A9, and had no obvious binding to other proteins in the S100 family ( Figure 2 ).

[0261] 2.5.3 Specificity of anti-S100A8 / A9 mAbs and other unrelated proteins

[0262] The binding specificity of anti-S100A8 / A9 mAbs to unrelated proteins was detected by ELISA. 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 the plate was blocked with blocking solution (4% milk-PBS) at 37°C for 1 hour, and anti-S100A8 / A9 monoclonal antibody was added, with a concentration of 20 μg / mL, and bound to the plate at 37°C for 1 hour, then washed 4 times with PBST, and then horseradish enzyme-labeled goat anti-human IgG (Zhongshan Jinqiao, ZB-2304) was added. The ELISA results showed that the anti-S100A8 / A9 monoclonal antibody prepared in this application specifically bound to hS100A8 / A9, and had no obvious binding to random unrelated proteins ( Figure 3 ).

[0263] 2.5.4 Species-specific analysis of anti-S100A8 / A9 mAbs

[0264] The species specificity of anti-S100A8 / A9 monoclonal antibody was detected by ELISA. Anti-S100A8 / A9 monoclonal antibody (0.5 μg / mL, 50 μL / well, coated overnight at 4°C) was coated on a 96-well plate. The liquid in the plate was discarded the next day, and the plate was blocked with blocking solution (4% milk-PBS) at 37°C for 1 hour. S100A8 / A9 antigens of three species (human heterodimeric antigen hS100A8 / A9, mouse heterodimeric antigen mS100A8 / A9, monkey heterodimeric antigen cS100A8 / A9) were added respectively. The initial concentration of antigen S100A8 / A9 was 20 μg / mL. A 5-fold gradient was set, with a total of 8 gradients. The plate was bound at 37°C for 1 hour, and then washed 4 times with PBST. The secondary antibody used was HRP-conjugated 6×His, His-Tag Monoclonal antibody (proteintech, Cat No: HRP-66005). The ELISA results showed that the anti-S100A8 / A9 monoclonal antibody screened in this application had the strongest affinity with hS100A8 / A9, followed by cS100A8 / A9, while the anti-S100A8 / A9 monoclonal antibody had no obvious binding with mS100A8 / A9 ( Figure 4 ).

[0265] 2.5.5 Stability analysis of anti-S100A8 / A9 mAbs in serum and PBS

[0266] 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 filtered and sterilized anti-S100A8 / A9 monoclonal antibody samples and dilute them to 100μg / mL with sterile PBS and mouse serum, respectively, with a volume of 200μL / tube, 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 method ( Figure 5A and 5B ). The results showed that the hS100A8 / A9 antibody obtained in the present application had good antigen binding stability in both C57BL / 6 mouse serum and PBS.

[0267] Example 3: Epitope Analysis of Anti-S100A8 / A9 Monoclonal Antibodies Binding to hS100A8 / A9

[0268] The binding epitopes of antigens and antibodies were analyzed by cryo-electron microscopy. The implementation method was summarized as follows: S100A8 / A9 samples were dropped onto a glow-treated grid. The grid specifications were Quantifoil Au300R1.2 / 1.3, and the photos 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 photos were collected, and the electron dose for each photo was The focal length range is -1.0μm--2.0μm. The 6258 good micrographs collected were further processed using cryoSMART (Shuimu Biotechnology Co., Ltd.). Blob-picker automatically picked particles, and then after two rounds of 2D classification, the best particles were selected as templates for Template picking for 3D reconstruction, and the final resolution was 2.54777777. Density map. AlphaFold2 prediction was performed based on the amino acid sequence, and the predicted results were matched with the calculated map using UCSFChimera 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 show ( Figure 6 ), the antigen core epitope is S100A8: D14, L21, G24, S100A9: E60, K72, D98, E99, P101, G102, H103. The antibody core binding site is light chain: K52, heavy chain: S54, S57, Y59, K99, R101, P102, R104.

[0269] Example 4: In vitro biological activity assay of anti-S100A8 / A9 mAbs

[0270] 4.1 Inhibitory effect of anti-S100A8 / A9 mAbs on the release of inflammatory factors from hS100A8 / A9 activated human PBMCs

[0271] The anti-S100A8 / A9 monoclonal antibody was prepared to a starting concentration of 2000 μg / mL and diluted 5 times in a gradient, and 8 dilutions were added to a 96-well plate (purchased from Corning, catalog number 3599), 50 μL per well. The hS100A8 / A9 protein was prepared at a concentration of 60 μg / mL, and 50 μL per well was added to a 96-well plate and mixed with the gradient concentrations of anti-S100A8 / A9 monoclonal antibodies. The antigen and antibody pyrogens used in this experiment were less than 2 EU / mL.

[0272] The ligand protein and gradient diluted antibody were mixed in equal volumes and incubated at room temperature for 30 min. Human PBMC cells (purchased from Maishun Biotechnology Co., Ltd., catalog number PB010C) were resuscitated and the cell density was adjusted to 1.0×10 6 cells / mL, 100 μL per well was plated in 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 set up at the same time, and the volume of the culture medium in the control group was supplemented. The cell plate was placed at 37°C CO 2 Incubate in an incubator for about 20 hours before testing. All drugs, 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 ThermoFisher, catalog number 10099141C). After the incubation, take the cell supernatant and perform a certain dilution. Use human IL-6 ELISA detection kit (purchased from Beijing Dakowei Biotechnology Co., Ltd., catalog number 1110603) and human TNF-α ELISA detection kit (purchased from Beijing Dakowei Biotechnology Co., Ltd., catalog number 1117203) to refer to the detection instructions, perform cytokine detection, read OD450, and draw a dose-effect curve. The results show that the anti-S100A8 / A9 monoclonal antibody in this application can inhibit the binding of hS100A8 / A9 to PBMC cell surface receptors and inhibit the inflammatory factor IL-6 ( Fig. 7A ), TNF-α( Figure 7B )’s release.

[0273] 4.2 Anti-S100A8 / A9 mAbs inhibit the proliferation and toxicity of hS100A8 / A9 on mouse lymphoma cells EL-4

[0274] 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 5 cells / mL, 100 μL per well, added to a 96-well plate, mixed evenly with human S100A8 / A9 protein, and incubated at 37°C CO 2Incubate in an incubator for 6 hours. Prepare the anti-S100A8 / A9 monoclonal antibody to a starting concentration of 9240 μg / mL, dilute the antibody 3-fold with antibody diluent, a total of 8 gradients, and add 50 μL to the corresponding position in the 96-well plate. All drug, antigen dilution and cell density adjustment were performed using DMEM complete medium (purchased from Thermo Fisher, catalog number C11995500BT) containing 10% horse serum (HS, purchased from EallBio, catalog number 03.16002A). At the same time, set up a blank group (EL-4only) and a negative control group (EL-4+S100A8 / A9), make up the volume of the culture medium in the control group, and place the cell plate at 37°C CO 2 After incubation for about 17 hours in the incubator, CCK-8 reagent (purchased from EallBio, catalog number 03.17002DC) was added, and OD450 was measured after incubation for about 4 hours to draw a dose-effect curve. The results showed that ( Figure 8 ), the hS100A8 / A9 antibody in the present application can bind to hS100A8 / A9, block the proliferation toxicity of hS100A8 / A9 on mouse lymphocytes EL-4, promote the proliferation of EL-4 cells, and show an obvious dose effect.

[0275] Example 5: Anti-S100A8 / A9 monoclonal antibody blocks the binding of hS100A8 / A9 to receptors

[0276] 5.1 Anti-S100A8 / A9 mAbs block the binding of hS100A8 / A9 to cell surface TLR4 receptors or RAGE receptors

[0277] Anti-S100A8 / A9 monoclonal antibody and isotype control antibody (IgG) were prepared to a starting concentration of 800 μg / mL, and the antibody was diluted 3-fold (to interact with TLR4 receptor) or 5-fold (to interact with RAGE receptor) with antibody diluent, with a total of 10 or 8 gradients, and added to a 96-well plate (purchased from Corning, catalog number 3894), 50 μL per well. Human S100A8 / A9 ligand protein was prepared to a concentration of 32 μg / mL (to interact with TLR4 receptor) or 8 μg / mL (to interact with RAGE receptor), 50 μL per well, and added to a 96-well plate. The ligand protein and the gradient diluted antibody were mixed evenly in equal volumes and incubated at room temperature for 30 minutes.

[0278] HEK293-TLR4 engineering cell line was used to detect the binding and blocking of hS100A8 / A9 to surface TLR4 receptors, and HEK293-RAGE engineering cell line was used to detect the binding and blocking of hS100A8 / A9 to surface RAGE receptors. Single cell suspension was obtained and counted, and resuspended in PBS buffer after centrifugation to adjust the cell density to 2.0×10 6cells / mL, 100 μL per well was added to the corresponding position of the 96-well plate, and incubated at 4°C for 1 hour. After incubation, the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. 200 μL PBS buffer was added to each well, and the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. The pre-prepared FITC anti-His-Tag antibody (purchased from Biolegend, catalog 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 the 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 interaction between hS100A8 / A9 and TLR4 on the surface of the engineered cell line ( Fig. 9A ) or RAGE( Fig. 9B ) showed a significant dose effect. No blocking effect was observed with the isotype control antibody.

[0279] 5.2 Anti-S100A8 / A9 mAbs block the binding of hS100A8 / A9 to PBMC cells and THP-1 cells

[0280] The anti-S100A8 / A9 monoclonal antibody was prepared to a starting concentration of 800 μg / mL, and the antibody was diluted 5-fold (for interaction with PBMC) or 4-fold (for interaction with THP-1 cells) with antibody diluent, with a total of 8 gradients, and added to a 96-well plate (purchased from Corning, catalog number 3894), 50 μL per well. The human S100A8 / A9 ligand protein was prepared at a concentration of 50 μg / mL, 50 μL per well, and added to a 96-well plate. The ligand protein and the gradient diluted antibody were mixed evenly in equal volumes and incubated at room temperature for 30 minutes.

[0281] PBMC cells and THP-1 cells were taken separately 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 of the 96-well plate, and incubated at 4°C for 1 hour. After incubation, the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. 200 μL PBS buffer was added to each well, and the cells were washed once by centrifugation at 3000 rpm, and the cell pellets were collected. The pre-prepared FITC anti-His-Tag antibody (purchased from Biolegend, catalog 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 resuspension in 200 μL PBS buffer, the cells were detected on the 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 with PBMC cells ( Fig. 10A ), and can also compete with hS100A8 / A9 for binding to THP-1 cells ( Fig. 10B ). There was a significant dose effect.

[0282] Example 6: Activity of anti-S100A8 / A9 monoclonal antibody genetically engineered mutants

[0283] 6.1 Binding activity of single-point mutations in the CDR region of anti-S100A8 / A9 monoclonal antibodies

[0284] Alanine scanning and single-site directed mutagenesis were performed on the six CDR regions to obtain single-point mutant antibodies, which were expressed and purified using the method described in 2.3 of Example 2, and the binding activities of the purified mutant antibodies and the parental antibodies were compared using the ELISA method described in 2.4 of Example 2. After screening, it was found that compared with the parental antibody (CDR sequences are shown in SEQ ID Nos: 1, 2, 3, 4, 5, and 6, respectively), the mutant antibodies shown in the following Table 3 all retained the binding activity of the parental antibody.

[0285] 6.2 Biological activity of single-point mutation antibodies in the CDR region of anti-S100A8 / A9 monoclonal antibodies

[0286] The method described in 2.3 of Example 2 was used to express and purify the 38 single-point mutant antibodies in Table 3 below, and the biological activity of the single-point mutant antibodies was reflected by detecting the activity of inhibiting hS100A8 / A9 from activating human PBMC cells to release IL-6. The specific detection method is as follows: Different mutants of anti-S100A8 / A9 monoclonal antibodies were prepared into antibody diluents with a starting concentration of 800 μg / mL and 5× gradient dilutions, with a total of 7 gradients, and added to a 96-well plate (purchased from Corning, item number 3799), 50 μL per well. The hS100A8 / A9 ligand protein was prepared at a concentration of 80 μg / mL, 50 μL per well, and added to a 96-well plate. The antigen and antibody pyrogens used in this experiment were all less than 2 EU / ml. The ligand protein and the gradient diluted antibody were mixed evenly 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 in the corresponding position of a 96-well plate and mixed evenly with the pre-incubation mixture. A blank control group (PBMC only) and a positive control group (PBMC+S100A8 / A9) were set up at the same time, and the volume of the culture medium in the control group was supplemented. The cell plate was placed at 37°C CO 2 Incubate in an incubator for about 20 hours before testing. All antibody 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, take the cell supernatant, dilute it to a certain extent, and use the human IL-6 ELISA detection kit (purchased from Beijing Dakoway Biotechnology Co., Ltd., catalog number 1110603) to refer to the detection instructions to detect cytokines, read OD450, and draw a dose-effect curve. The results show ( Fig.11 and Table 3), the 38 mutants of the anti-S100A8 / A9 monoclonal antibody in the present application (as shown in Table 3 below) can inhibit the binding of hS100A8 / A9 to PBMC cell surface receptors and the release of IL-6, showing a significant dose-dependent effect.

[0287] The core sites of the HCDR region are F29, Y32, A33, M34, Y59, A61, D62, Y100, S103, V105, F106, and S108. The cytological activity of the core amino acid sites increased by about 10 times after mutation: F29Y, F29W, Y59W, A61V, A61H, V105A, and F106Y. The cytological activity of the sites increased by about 2 times after mutation: F29H, A33V, Y100F, Y100H, and F106H. The antibodies obtained after mutation of other sites also retained good cytological activity: 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.

[0288] The core sites of LCDR region are Y31, E49, D50, S51, K52, A92, Q88. After mutation of the core amino acid sites, the cytological activity is increased by about 5 times: A92I, the cytological activity is increased by about 2 times after site mutation: Y31F, E49Q, the cytological activity is increased by less than 2 times after site mutation: K52H, A92L, and the antibodies obtained after mutation of other sites also retain good cytological activity: D50Q, D50H, K52P, Q88D, S51A, S51H, K52R, A92V, S94A. The sequences of the LCDR regions of antibodies with good in vitro biological activity are shown in Table 3 below.

[0289] Table 3 Changes in antibody cytological activity after single-point mutation of CDR

[0290]

[0291]

[0292] 6.3 Binding activity of anti-S100A8 / A9 monoclonal antibody CDR region combined mutant antibodies

[0293] Combined with the results of Examples 6.1 and 6.2, the CDR region was subjected to combined mutations at multiple mutation sites to obtain a mutant antibody with combined mutations, the combined mutant antibody was expressed and purified using the method described in 2.3 of Example 2, and the binding activity of the purified combined mutant antibody and the parent antibody was compared 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, 6, respectively), the combined mutant antibodies shown in Table 4 below all retained the binding activity of the parent antibody.

[0294] 6.4 Biological activity of anti-S100A8 / A9 monoclonal antibody CDR region combination mutant antibodies

[0295] The method described in 2.3 of Example 2 was used to express and purify the 80 multi-point combination mutant antibodies in Table 4 below, and the biological activity of the multi-point combination mutant antibodies was detected by referring to the method described in 6.2 of Example 6. The results showed (Figure 12 and Table 4) that the 80 combination mutants of the anti-S100A8 / A9 monoclonal antibodies in the present application (as shown in Table 4 below) were able to inhibit the binding of hS100A8 / A9 to the PBMC cell surface receptors and inhibit the release of IL-6, showing a significant dose-dependent effect.

[0296] 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.

[0297] 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.

[0298] Table 4 Changes in antibody cytological activity after CDR combination mutation

[0299]

[0300]

[0301]

[0302] Example 7: Pharmacokinetics of anti-S100A8 / A9 monoclonal antibodies

[0303] In order to investigate the metabolism of the anti-S100A8 / A9 monoclonal antibody of the present application in mice, 5 male C57BL / 6 mice were selected and administered by tail vein injection at a dose of 10 mg / kg. The antibody concentration in serum was detected by taking blood at different time points, and the PK blood collection points were 5 minutes, 2 hours, 4 hours, 8 hours, 1 day, 3 days, 7 days and 14 days after administration.

[0304] The total antibody and active antibody concentrations of serum samples at the drug-generation blood collection point were detected by ELISA. Goat anti-human Fc antibody (1:2000, Sigma-Aldrich, Product number: I8885) and antigen (1μg / mL, hS100A8 / A9) were diluted with PBS, 100μL / well, and coated overnight at 4℃. 3% skim milk powder (diluted with PBS) was added to the well plate, 300μL / well, and blocked at 37℃ for 2h. Antibody standards and mouse serum with known concentrations were added to the well plate, 100μL / well, and incubated at 37℃ for 1h. Wash the plate, add horseradish enzyme-labeled goat anti-human IgG (H+L) (1:5000, Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., No.: ZB-2304), 100μL / well, and incubate at 37℃ for 45min. 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 an ELISA reader, and 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.

[0305] The results show that Fig.13 ), 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.

[0306] Example 8: Effect of anti-S100A8 / A9 monoclonal antibody on acute myocardial infarction and myocardial ischemia / reperfusion in S100A8 / A9 humanized mice Improvement of cardiac injury and cardiac function in patients with perfusion injury

[0307] 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

[0308] In order to investigate the dose-dependent improvement effect 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 4 groups, namely, IgG negative control group, anti-S100A8 / A9 monoclonal antibody low, medium and high dose groups, 10 mice in each group, 1 / 2 of males and females, and the left coronary artery was ligated to establish an acute myocardial infarction model. Some mice died during the operation, and 8 mice were finally administered in each group. Immediately after the myocardial infarction surgery, anti-S100A8 / A9 monoclonal antibody or IgG control was administered by medial canthal vein injection. 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 dosage was 24 mg / kg. The state of the mice was observed daily. The cardiac function of the mice was detected by echocardiography 14 days after myocardial infarction, and the left ventricular ejection fraction was calculated. The mice were killed, and the heart tissues were obtained, fixed, paraffin-embedded, sectioned, and stained with Masson's staining. The infarct area was measured and calculated.

[0309] The results showed that low, medium and high doses of anti-S100A8 / A9 monoclonal antibody 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 left ventricular ejection fraction of the high dose group of anti-S100A8 / A9 monoclonal antibody was increased compared with the low dose group, and the difference was statistically significant (p<0.05) ( Fig.14 ).

[0310] Compared with the IgG negative control group, the low, medium and high doses of anti-S100A8 / A9 monoclonal antibody could reduce the myocardial infarction area, and the difference was statistically significant (p<0.05). Compared with the low dose group, the high dose group of anti-S100A8 / A9 monoclonal antibody had a smaller myocardial infarction area, and the difference was statistically significant (p<0.05) ( Fig.15 ).

[0311] 8.2 Effects of different administration times of anti-S100A8 / A9 monoclonal antibodies on the improvement of cardiac injury and cardiac function in S100A8 / A9 humanized mice with acute myocardial infarction

[0312] In order to explore the administration time window of the anti-S100A8 / A9 monoclonal antibody of the present application for improving acute myocardial infarction in mice, 50 hS100A8 / A9 humanized mice were selected and randomly divided into 5 groups, 1 / 2 of males and 1 / 2 of females, respectively, IgG negative control group, anti-S100A8 / A9 monoclonal antibody administration group 2 hours (h), 4h, 8h, 10h after surgery, all left coronary arteries were ligated, and acute myocardial infarction models were established. Some mice died during the myocardial infarction surgery, and the remaining IgG group had 9 mice, 7 mice in the 2h and 4h groups, and 8 mice in the 8h and 10h groups. The IgG negative control group was given IgG control by medial canthal vein injection 2 hours after myocardial infarction surgery, with a dose of 20 mg / kg, and the other 4 groups were given anti-S100A8 / A9 monoclonal antibodies at 2, 4, 8, and 10 hours after surgery, respectively, with a dose of 20 mg / kg. 14 days after myocardial infarction, the cardiac function of mice was detected by echocardiography, and the left ventricular ejection fraction was calculated. The mice were killed, and the heart tissues were obtained, fixed, paraffin-embedded, sliced, and stained with Masson's to measure and calculate the infarct area.

[0313] The results showed that the 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 could not improve the ejection fraction compared with the IgG negative control group, and the group administered 2 hours after surgery had a statistically significant improvement in the left ventricular ejection fraction compared with the 10 hours group (p<0.05) ( Fig.16 ).

[0314] Anti-S100A8 / A9 monoclonal antibody administration at 2, 4, and 8 hours after surgery can reduce the myocardial infarction area of ​​mice compared with the IgG negative control group, and the difference is statistically significant (p<0.05); there is no significant improvement in the myocardial infarction area in the 10-hour group compared with the IgG negative control group; the myocardial infarction area in the 2-hour and 4-hour groups after surgery is reduced compared with the 8-hour group, and the difference is statistically significant (p<0.05); the myocardial infarction area in the 2-hour and 4-hour groups after surgery is reduced compared with the 10-hour group, and the difference is statistically significant (p<0.05) ( Fig.17 ).

[0315] The results showed that the therapeutic time window for administration of anti-S100A8 / A9 monoclonal antibodies after myocardial infarction is within 8 hours.

[0316] 8.3 Effect of anti-S100A8 / A9 monoclonal antibody on the improvement of cardiac function after myocardial ischemia / reperfusion in S100A8 / A9 humanized mice

[0317] In order to investigate the effect of the anti-S100A8 / A9 monoclonal antibody of the present application on the improvement of cardiac function after myocardial infarction / vascular recanalization in mice, 20 S100A8 / A9 humanized mice were selected and randomly divided into 2 groups, IgG negative control group and anti-S100A8 / A9 monoclonal antibody treatment group, 10 mice in each group, 1 / 2 of male and female, all of which were ligated for 60 minutes on the left coronary artery, and then the ligation was untied, and the blood vessels were recanalized, simulating the recanalization of blood vessels after thrombolysis or PCI surgery after myocardial infarction. During the operation, some mice died or the modeling was unsuccessful, and 7 mice remained in the IgG negative control group, and 9 mice in the anti-S100A8 / A9 monoclonal antibody group were administered. Anti-S100A8 / A9 monoclonal antibody or IgG was given immediately after vascular recanalization, and the dosage was 20 mg / kg. The state of the mice was observed every day after surgery. The cardiac function of the mice was detected by echocardiography 28 days after administration, and the left ventricular ejection fraction was calculated.

[0318] The results showed that anti-S100A8 / A9 monoclonal antibody could significantly increase the left ventricular ejection fraction of mice after myocardial ischemia / reperfusion (p<0.05) and improve cardiac function ( Fig.18 ).

[0319] 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 implementations have been described, for the applicant or other persons skilled in the art, there may be or are currently no foreseeable substitutions, modifications, changes, improvements, and substantial equivalents of the above-mentioned implementations. Therefore, the attached claims submitted and the claims that may be amended are intended to cover all such substitutions, modifications, changes, improvements, and substantial equivalents. It is important that with the evolution of technology, 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 an 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 is 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 is 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 is 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, 35-48; The heavy chain CDR3 is selected from at least one of the amino acid sequences shown in SEQ ID NOs: 3, 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, 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 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 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 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 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 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 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 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 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 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 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 SEQ ID NOs: 1, 2, 57, 4, 5, and 6, respectively; (26) 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, 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 SEQ ID NOs: 1, 2, 3, 4, 70, and 6, respectively; (28) 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, 71, and 6, respectively; (29) 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, 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 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 SEQ ID NOs: 1, 2, 3, 4, 74, and 6, respectively; (32) 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, 75, and 6, respectively; (33) 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, 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 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 SEQ ID NOs: 1, 2, 3, 4, 5, and 96, respectively; (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; (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 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 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 SEQ ID NOs: 1, 42, 57, 4, 73, and 6, respectively; (44) the 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, 57, 4, 73, and 100, respectively; (45) 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, 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 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) 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, 66, 5, and 99, respectively; (49) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are amino acid sequences represented by SEQ ID NOs: 1, 38, 57, 4, 73, and 99, respectively; (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 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 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 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 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 SEQ ID NOs: 1, 42, 57, 66, 73, and 104, respectively; (58) 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, 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 SEQ ID NOs: 1, 42, 64, 4, 73, and 104, respectively; (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 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 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 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 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 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 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 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 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 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 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 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 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 SEQ ID NOs: 1, 42, 57, 4, 89, and 100, respectively; (79) 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, 42, 57, 4, 90, and 100, respectively; (80) the heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR1, and light chain CDR3 are represented by the amino acid sequences of 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 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 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 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 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 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 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 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 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 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 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 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 SEQ ID NOs: 1, 45, 57, 69, 73, and 100, respectively; (114) 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, 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) 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, 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 SEQ ID NOs: 27, 42, 58, 4, 73, and 108, respectively; (118) the 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: 27, 42, 60, 4, 73, and 108, respectively; (119) 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: 27, 42, 61, 4, 73, and 108, respectively.

4. The antibody or antigen-binding fragment thereof according to claim 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 4 , 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 double-chain antibody, a three-chain antibody, 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.

7. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein the heavy chain 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 sites in the sequence shown in SEQ ID NO: 11, wherein the mutation comprises insertion, deletion and / or substitution.

8. The antibody or antigen-binding fragment thereof according to claim 7, wherein the mutation is a substitution, and the substituted amino acid site 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 5, wherein the light chain variable region 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 sites in the sequence shown in SEQ ID NO: 12, wherein the mutation comprises insertion, deletion and / or substitution.

10. The antibody or antigen-binding fragment thereof according to claim 9, wherein the mutation is a substitution, and the substituted amino acid site 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: It 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 radioactive isotope, a structural part that prolongs the half-life, a detectable marker 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 marker 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.

21. The use according to claim 20, wherein the disease is associated with an increased expression level 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 microcirculation 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 of any one of claims 1 to 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.

25. The method of claim 24, wherein the disease is associated with elevated expression levels of oligomers, dimers, tetramers, or multimers greater than tetramers 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 area, 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 to 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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