Antibody of anti-S100 protein and application thereof

Abstract

The invention discloses an anti-S100 protein antibody and application thereof, and relates to the field of antibodies. The anti-S100 protein antibody disclosed by the invention comprises a heavy chain complementarity determining region and a light chain complementarity determining region, provides an important raw material source for detection of S100 protein, and has good activity and sensitivity.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202311433224.7 filed with the Chinese Patent Office on October 31, 2023, entitled “An antibody against S100 protein and its application”, the entire contents of which are incorporated by reference into this application. Technical Field

[0003] The present invention relates to the field of antibody technology, and in particular to an antibody against S100 protein and an application thereof. Background Art

[0004] S100 protein was discovered by Moore in bovine brain in 1965. It was named because it can be dissolved in 100% ammonium sulfate solution. S100 protein is an acidic calcium-binding protein, composed of 21 members, with a relative molecular mass of 10,000-21,000. Its gene is located on chromosome 1q21 and belongs to the EF-hand motif superfamily. Each S100 protein has two calcium ion binding regions (α helix-loop-α helix) with EF-hand structure. The calcium ion binding region near the C-terminus has high affinity with calcium ions, while the calcium ion binding region near the N-terminus has the specificity of the S100 family itself. The intermediate hinge region and the C-terminal extension region between the two are different among different members, so they are responsible for their specific biological characteristics.

[0005] S100 protein is widely expressed in normal tissue cells derived from neuroectoderm, mesoderm, and ectoderm, such as glial cells, Schwann cells, melanocytes, mammary epithelial or myoepithelial cells, or tumors originating from these cells. Most of the S100 protein appears in cells and participates in the regulation of intracellular proliferation, differentiation, apoptosis, calcium homeostasis, energy metabolism, migration and other functions. When S100 protein is overexpressed in cells, it is released into the blood. Under normal circumstances, S100 protein is mainly metabolized in the kidneys, with a biological half-life of about 2 hours. The content in normal adult serum is less than 0.2ug / L, which is usually difficult to detect. It is considered pathological if it is greater than 0.5ug / L.

[0006] S100 protein can specifically bind to calcium ions and regulate the interaction between proteins by activating multiple enzymes, playing an important role in cell cycle growth and differentiation. Studies have found that 15 member genes of the S100 family are located in the long arm of chromosome 1, region 2, band 1 (1q21), which has poor stability. Tumor genes in this region often recombine frequently, which can easily cause uncontrolled expression of S100 genes. Disorders in the expression of S100 proteins are associated with many malignant tumors. Therefore, the S100 family is closely related to the occurrence and development of tumors. Studies have confirmed that S100 protein and P53 tumor suppressor protein interact at the nucleic acid level, inhibiting protein kinase phosphorylation, thereby affecting the regulation of cell growth. S100 protein can also change the adhesion of cells, making it easier for cancer cells to attach to tissues and cause clonal proliferation. The value of S100 protein serological detection in the diagnosis of central nervous system damage has been clinically recognized, and its value as a tumor marker has gradually attracted attention and experimental recognition. According to relevant research reports, the expression of S100 protein is significantly upregulated in invasive hepatocellular carcinoma, and it is believed that S100 protein may become a valuable molecular marker for liver cancer classification and invasion prediction. S100 protein is closely related to the occurrence of non-small cell lung cancer, etc. The expression of S100 protein is also significantly upregulated in esophageal cancer, colon cancer and malignant melanoma. With the deepening of research on S100 protein, S100 protein is also closely related to human heart disease, oral cancer, breast cancer and prostate cancer, mainly manifested in that their protein expression levels are significantly abnormal when the disease occurs, which is of great significance for clinical diagnosis. Therefore, in the current clinical and scientific research fields, accurate, convenient and fast detection methods for S100 protein are particularly important.

[0007] Traditional detection methods for S100 protein mainly include electrochemiluminescence, radioimmunoassay, immunoradiometric assay and fluorescent immunoassay. The electrochemiluminescence method has high accuracy in quantitative detection and has the advantages of quantitativeness and stability, but it is not suitable for modern automatic immunoassay instruments and requires the use of special supporting instruments. The investment cost is high, so it is difficult to promote and apply. Radioimmunoassay and immunoradiometric assay have the disadvantages of radioactive pollution and hazards, short half-life of nuclides, short stability period, and cumbersome operation, which limit their application. Fluorescent immunoassay requires special fluorescent equipment and is not easy to popularize clinically.

[0008] At present, the detection methods for S100 protein on the market mainly include enzyme-linked immunosorbent assay (ELISA), chemiluminescence, flow cytometry, time-resolved fluorescence immunoassay, photochemiluminescence immunoassay, colloidal gold, etc. These detection methods are simple to operate, fast and accurate, and do not require complex equipment. These different detection methods have their own advantages and disadvantages, but they all require antibodies against S100 protein.

[0009] Therefore, there is a strong demand in the art for anti-S100 protein antibodies with good performance. Summary of the invention

[0010] The present application provides an anti-S100 protein antibody, which provides an important source of raw materials for the detection of S100 protein and has good activity and sensitivity.

[0011] In order to achieve the above-mentioned object, according to one aspect of the present invention, an anti-S100 protein antibody is provided, which comprises three complementarity determining regions of a heavy chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33, and three complementarity determining regions of a light chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45.

[0012] In order to achieve the above object, according to a second aspect of the present invention, an anti-S100 protein antibody is provided, wherein the antibody comprises the following complementary determining regions:

[0013] HCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:1 (NYWIH);

[0014] HCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:2 (GEINPSNGRTKFNEKFKT);

[0015] HCDR3 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 3 (RFFIASIGGAMDY), 17 (RFFLASIGGAMDY), 18 (RFFIASLGGAMDY) or 19 (RFFLASLGGAMDY);

[0016] LCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:4 (SASSSVSSSYLY);

[0017] LCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:5 (GTSNLAS); and

[0018] LCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO:6 (HQWSSYPPT).

[0019] In order to achieve the above-mentioned object, according to the third aspect of the present invention, there is provided an anti-S100 protein antibody, comprising a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33; and the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45.

[0020] In order to achieve the above-mentioned object, according to the fourth aspect of the present invention, there is provided an anti-S100 protein antibody, comprising a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 34, 35, 36, 37, 38, 39, and 40; and the amino acid sequence of the light chain is shown in any one of SEQ ID NOs: 46, 47, 48, 49, and 50.

[0021] In order to achieve the above object, according to the fifth aspect of the present invention, an antibody conjugate is provided, wherein the antibody conjugate comprises the above antibody.

[0022] In order to achieve the above object, according to the sixth aspect of the present invention, a reagent or a kit is provided, wherein the reagent or the kit comprises the above antibody or the above antibody conjugate.

[0023] In order to achieve the above object, according to an eighth aspect of the present invention, there is provided a use of the above antibody, antibody conjugate, reagent or kit in the preparation of a product for detecting S100 protein.

[0024] In order to achieve the above object, the present invention also provides a nucleic acid, a vector, a cell and a method for preparing the above antibody. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0026] Figure 1 These are the results of reducing SDS-PAGE of Anti-S100 4G8 Rmb1 to Anti-S100 4G8 Rmb11. DETAILED DESCRIPTION

[0027] In a first aspect, an embodiment of the present invention provides an anti-S100 protein antibody, comprising three complementary determining regions of a heavy chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33, and three complementary determining regions of a light chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45.

[0028] It should be noted that HCDR1, HCDR2 and HCDR3 are amino acid sequences consistent with HCDR1, HCDR2 and HCDR3 of the same heavy chain variable region defined in the antibody described in the first aspect, and LCDR1, LCDR2 and LCDR3 are amino acid sequences consistent with LCDR1, LCDR2 and LCDR3 of the same light chain variable region defined in the antibody described in the first aspect.

[0029] For example, the HCDR1, HCDR2, and HCDR3 are amino acid sequences consistent with HCDR1, HCDR2, and HCDR3 of the heavy chain variable region shown in SEQ ID NO:27; the LCDR1, LCDR2, and LCDR3 are amino acid sequences consistent with LCDR1, LCDR2, and LCDR3 of the light chain variable region shown in SEQ ID NO:41.

[0030] In the present invention, the term "antibody" is used in the broadest sense, and may include full-length monoclonal antibodies, bispecific antibodies, multispecific antibodies, chimeric antibodies, or antigen-binding fragments of antibodies, as long as they exhibit the desired antigen-binding activity. Antibody antigen-binding fragments generally have the same binding specificity as the antibody from which they are derived, and may be selected from any one of F(ab')2, Fab', Fab, Fv, and scFv. It is easy for a person skilled in the art to understand based on the contents described in the present invention that they can be obtained by methods such as enzymatic digestion (including pepsin or papain) and / or by chemical reduction to split disulfide bonds. Based on the structure of the complete antibody disclosed in the present invention, a person skilled in the art can easily obtain the antigen-binding fragment of the above-mentioned antibody.

[0031] Antigen-binding fragments of antibodies can also be synthesized by recombinant genetic techniques also known to those skilled in the art or by, for example, an automatic peptide synthesizer, such as those sold by Applied BioSystems and the like.

[0032] In the present invention, the term "complementarity determining region", "CDR" or "CDRs" refers to the highly variable region of the heavy and light chains of immunoglobulins, and refers to the region containing one or more or even all of the major amino acid residues that play a role in the binding of an antibody or antigen-binding fragment to its recognized antigen or epitope. In a specific embodiment of the present invention, CDRs refers to the highly variable region of the heavy and light chains of the antibody.

[0033] In the present invention, the heavy chain complementarity determining region is represented by HCDR, which includes HCDR1, HCDR2 and HCDR3; the light chain complementarity determining region is represented by LCDR, which includes LCDR1, LCDR2 and LCDR3.

[0034] The definition methods of CDR are well known in the art, and the CDR definition methods include: Kabat definition, Chothia definition, IMGT definition, Contact definition and AbM definition. As described herein, "Kabat definition" refers to the definition system described in Kabat et al., US Pept. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983). "Chothia definition" refers to Chothia et al., J Mol Biol 196: 901-917 (1987). There are other CDR definition methods that may not strictly follow one of the above schemes, but will still overlap with at least a portion of the CDR region defined by Kabat, although they may be shortened or extended based on predictions or experimental results of specific residues or residue groups. Exemplary defined CDRs are listed in Table 1 below, and the definitions in different documents are slightly different. Given the variable region amino acid sequence of an antibody, a person skilled in the art can routinely determine which residues contain a specific CDR. It should be noted that CDRs defined by other methods not limited to Table 1 also belong to the scope of protection of the present disclosure.

[0035] Table 1: CDR Definition 1

[0036] CDR Kabat AbM2 IMGT Chothia HCDR1 <![CDATA[H31~H35 3 ]]> <![CDATA[H26~H35 3 ]]> <![CDATA[H26~H33..5 5 ]]> <![CDATA[H26~H32..34 4 ]]> HCDR2 H50~H65 H50~H58 H51~H57 H52~H56 HCDR3 H95~H102 H95~H102 H93~H102 H95~H102 LCDR1 L24~L34 L24~L34 L27~L32 L24~L34 LCDR2 L50~L56 L50~L56 L50~L51 L50~L56 LCDR3 L89~L97 L89~L97 L89~L97 L89~L97

[0037] 1The numbering of all CDR definitions in Table 1 is based on the Kabat numbering system (see below), with the amino acid numbering on the heavy chain represented by "H+number" and the amino acid numbering on the light chain represented by "L+number". One of ordinary skill in the art can unambiguously map the Kabat numbering system to any variable region sequence without relying on any experimental data other than the sequence itself. As used herein, "Kabat numbering" refers to the numbering system described by Kabat et al., U.S. Patent. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983).

[0038] 2 "AbM" as used in Table 1 with a lowercase "b" refers to CDRs defined by Oxford Molecular's "AbM" antibody modeling software.

[0039] 3 If both H35A and H35B are absent, CDR-H1 ends at position 35; if only H35A is present, CDR-H1 ends at position 35A; if both H35A and H35B are present, CDR-H1 ends at position 35B.

[0040] 4 If both H35A and H35B are absent, CDR-H1 ends at position 32; if only H35A is present, CDR-H1 ends at position 33; if both H35A and H35B are present, CDR-H1 ends at position 34.

[0041] 5 If both H35A and H35B are absent, CDR-H1 ends at position 33; if only H35A is present, CDR-H1 ends at position 34; if both H35A and H35B are present, CDR-H1 ends at position 35.

[0042] According to an embodiment of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 is defined by any one system or a combination of multiple systems of Kabat, Chothia, IMGT, AbM or Contact.

[0043] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Kabat system.

[0044] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Chothia system.

[0045] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the IMGT system.

[0046] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the AbM system.

[0047] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a Contact system.

[0048] In some optional embodiments of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a combination of Kabat, Chothia, IMGT, AbM or Contact systems.

[0049] According to an embodiment of the present invention, the Kabat numbering positions corresponding to the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 defined by the Kabat, Chothia, AbM or IMGT systems are as follows:

[0050] CDR Kabat AbM IMGT Chothia HCDR1 H31~H35 H26~H35 H26~H33 H26~H32 HCDR2 H50~H65 H50~H58 H51~H57 H52~H56 HCDR3 H95~H102 H95~H102 H93~H102 H95~H102 LCDR1 L24~L34 L24~L34 L27~L32 L24~L34 LCDR2 L50~L56 L50~L56 L50~L51 L50~L56 LCDR3 L89~L97 L89~L97 L89~L97 L89~L97

[0051] In a second aspect, an embodiment of the present invention provides an antibody against S100 protein, wherein the antibody comprises the following complementary determining regions:

[0052] HCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:1 (NYWIH);

[0053] HCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:2 (GEINPSNGRTKFNEKFKT);

[0054] HCDR3 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 3 (RFFIASIGGAMDY), 17 (RFFLASIGGAMDY), 18 (RFFIASLGGAMDY) or 19 (RFFLASLGGAMDY);

[0055] LCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:4 (SASSSVSSSYLY);

[0056] LCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:5 (GTSNLAS); and

[0057] LCDR3 comprises or consists of the amino acid sequence shown in SEQ ID NO:6 (HQWSSYPPT).

[0058] According to an embodiment of the present invention, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Kabat system.

[0059] In the present invention, the "framework region" or "FR" region includes the heavy chain framework region and the light chain framework region, and refers to the region other than the CDR in the heavy chain variable region and the light chain variable region of the antibody; wherein the heavy chain framework region can be further subdivided into adjacent regions separated by CDR, including HFR1, HFR2, HFR3 and HFR4 framework regions; the light chain framework region can be further subdivided into adjacent regions separated by CDR, including LFR1, LFR2, LFR3 and LFR4 framework regions.

[0060] In the present invention, the heavy chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4; the light chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4.

[0061] In an alternative embodiment, the antibody of the first aspect or the second aspect further has HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4.

[0062] In an alternative embodiment, the HFR1 comprises / is SEQ ID NO:7 or an amino acid sequence having at least 80% identity thereto;

[0063] The HFR2 comprises / is SEQ ID NO:8 or an amino acid sequence having at least 80% identity thereto;

[0064] The HFR3 comprises / is SEQ ID NO:9 or an amino acid sequence having at least 80% identity thereto;

[0065] The HFR4 comprises / is SEQ ID NO: 10 or an amino acid sequence having at least 80% identity thereto;

[0066] The LFR1 comprises / is SEQ ID NO:11 or an amino acid sequence having at least 80% identity thereto;

[0067] The LFR2 comprises / is SEQ ID NO: 12 or an amino acid sequence having at least 80% identity thereto;

[0068] The LFR3 comprises / is SEQ ID NO: 13 or an amino acid sequence having at least 80% identity thereto; and

[0069] The LFR4 comprises / is SEQ ID NO: 14 or an amino acid sequence having at least 80% identity thereto.

[0070] It should be noted that, in other embodiments, the amino acid sequences of the framework regions of the anti-S100 protein antibodies provided by the present invention may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the above-mentioned corresponding framework regions (SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14).

[0071] In an optional embodiment, the HFR1 comprises / is represented by the amino acid sequence shown in SEQ ID NO:20.

[0072] In an optional embodiment, the HFR2 comprises / is represented by the amino acid sequence shown in SEQ ID NO:21.

[0073] In an optional embodiment, the HFR3 comprises / is represented by the amino acid sequence shown in SEQ ID NO:22.

[0074] In an optional embodiment, the LFR1 comprises / is an amino acid sequence as shown in SEQ ID NO:23, 24 or 25.

[0075] In an optional embodiment, the LFR2 comprises / is represented by the amino acid sequence shown in SEQ ID NO:26.

[0076] In a third aspect, an embodiment of the present invention provides an anti-S100 protein antibody, comprising a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33, and the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45.

[0077] In an optional embodiment, the heavy chain variable region and the light chain variable region described in the first aspect or the third aspect are selected from any combination of the following:

[0078]

[0079]

[0080] In an optional embodiment, the antibody described in the first aspect, the second aspect, and the third aspect further comprises a constant region.

[0081] In an alternative embodiment, the constant region includes a heavy chain constant region and / or a light chain constant region.

[0082] In an optional embodiment, the heavy chain constant region is selected from any one of the heavy chain constant regions of IgG, IgA, IgM, IgE, and IgD, or a combination of multiple constant region segments.

[0083] In an alternative embodiment, the heavy chain constant region includes CH1 of IgG, a hinge region of IgG, CH2 of IgM, CH3 of IgM and / or CH4 of IgM.

[0084] In an alternative embodiment, the IgG is selected from IgG1, IgG2, IgG3 or IgG4.

[0085] In an alternative embodiment, the light chain constant region is selected from a kappa-type or a lambda-type light chain constant region.

[0086] In an alternative embodiment, the species of origin of the constant region is cattle, horses, dairy cows, pigs, sheep, rats, mice, dogs, camels, cats, rabbits, donkeys, deer, minks, chickens, ducks, geese, turkeys, fighting cocks or humans.

[0087] In an alternative embodiment, the species origin of the constant region is mouse.

[0088] In an optional embodiment, the heavy chain constant region sequence (CH) is as shown in SEQ ID NO:15, and the light chain constant region (CL) sequence is as shown in SEQ ID NO:16.

[0089] It should be noted that, in other embodiments, the constant region sequence may have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the above-mentioned constant region (SEQ ID NO: 15 or 16).

[0090] In an alternative embodiment, the antibody comprises any one of F(ab)2, F(ab')2, Fab', Fab, Fv and scFv.

[0091] In a fourth aspect, the present invention provides an anti-S100 protein antibody, comprising a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 34, 35, 36, 37, 38, 39, and 40, and the amino acid sequence of the light chain is shown in any one of SEQ ID NOs: 46, 47, 48, 49, and 50.

[0092] In an optional embodiment, the antibody of the first aspect, the second aspect, the third aspect or the fourth aspect comprises any combination of the following heavy chains and light chains:

[0093]

[0094]

[0095] In a fifth aspect, the present invention provides an antibody conjugate, wherein the antibody conjugate comprises the above-mentioned antibody.

[0096] In an optional embodiment, the above-mentioned antibody conjugate further comprises biotin or a biotin derivative conjugated to the antibody.

[0097] In an optional embodiment, the antibody conjugate further comprises a label coupled to the antibody.

[0098] In an optional embodiment, the above-mentioned marker refers to a class of substances with properties that can be directly observed by the naked eye or detected or detected by an instrument, such as luminescence, color development, radioactivity, etc., through which qualitative or quantitative detection of the corresponding target can be achieved.

[0099] In an alternative embodiment, the label includes but is not limited to fluorescent dyes, enzymes, radioactive isotopes, chemiluminescent agents and nanoparticle labels.

[0100] In actual use, those skilled in the art can select a suitable marker according to the detection conditions or actual needs. No matter which marker is used, it belongs to the protection scope of the present invention.

[0101] In an optional embodiment, the fluorescent dye is not limited to fluorescein dyes and their derivatives (for example, including but not limited to fluorescein isothiocyanate (FITC), hydroxyfluorescein (FAM), tetrachlorofluorescein (TET), etc. or their analogs), rhodamine dyes and their derivatives (for example, including but not limited to red rhodamine (RBITC), tetramethylrhodamine (TAMRA), rhodamine B (TRITC), etc. or their analogs), Cy series dyes and their derivatives (for example, including but not limited to Cy2, Cy3, Cy3B, Cy3.5, Cy3.7, Cy3.8, Cy3.9, Cy3.10, Cy3.11, Cy3.12, Cy3.13, Cy3.14, Cy3.15, Cy3.16, Cy3.17, Cy3.18, Cy3.19, Cy3.20, Cy3.21, Cy3.22, Cy3.23, Cy3.24, Cy3.25, Cy3.26, Cy3.27, Cy3.28, Cy3.29, Cy3.30, Cy3.31, Cy3.32, Cy3.33, Cy3.34, Cy3.35, Cy3.36, Cy3.37, Cy3.38, Cy3.39, Cy3.40, Cy3.41, Cy3.42, Cy3.43, Cy3.44, Cy3.45, Cy3.46, Cy3.47, Cy3.48, Cy3.49, Cy3.50, Cy3.51, Cy3.52, Cy3.53, Cy3.54, Cy3.55, Cy3.56, Cy3.57, Cy3.58, Cy3.59, Cy3.59, Cy3.59, Cy3.59, Cy3.59, Cy3.5 y5, Cy5.5, Cy3, etc. or their analogs), Alexa series dyes and their derivatives (for example, including but not limited to AlexaFluor350, 405, 430, 488, 532, 546, 555, 568, 594, 610, 33, 647, 680, 700, 750, etc. or their analogs) and protein dyes and their derivatives (for example, including but not limited to phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC), pre-chlorophyll protein (preCP), etc.).

[0102] In alternative embodiments, the enzyme includes, but is not limited to, horseradish peroxidase, alkaline phosphatase, β-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and 6-phosphoglucose deoxygenase.

[0103] In an optional embodiment, the radioactive isotopes include but are not limited to 212Bi, 131I, 111In, 90Y, 186Re, 211At, 125I, 188Re, 153Sm, 213Bi, 32P, 94mTc, 99mTc, 203Pb, 67Ga, 68Ga, 43Sc, 47Sc, 110mIn, 97Ru, 62Cu, 64Cu, 67Cu, 68Cu, 86Y, 88Y, 121Sn, 161Tb, 166Ho, 105Rh, 177Lu, 172Lu and 18F.

[0104] In an optional embodiment, the chemiluminescent reagent includes but is not limited to luminol and its derivatives, lucigenin, crustacean fluorescein and its derivatives, bipyridine ruthenium and its derivatives, acridinium esters and their derivatives, dioxetanes and their derivatives, lophanes and their derivatives, and peroxyoxalates and their derivatives.

[0105] In an optional embodiment, the nanoparticle markers include, but are not limited to, nanoparticles, colloids, organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles, and rare earth complex nanoparticles.

[0106] In alternative embodiments, the colloid includes, but is not limited to, colloidal metals, colloidal carbon, disperse dyes, dye-labeled microspheres, and latex.

[0107] In an alternative embodiment, the colloidal metal includes, but is not limited to, colloidal gold, colloidal silver, and colloidal selenium.

[0108] In an optional embodiment, the colloidal metal is colloidal gold.

[0109] In an optional embodiment, the above-mentioned antibody conjugate further includes a solid phase carrier coupled to the antibody.

[0110] In an alternative embodiment, the solid support is selected from microspheres, plates and membranes.

[0111] In an optional embodiment, the solid phase carrier includes but is not limited to magnetic microspheres, plastic microspheres, plastic microparticles, microwell plates, glass, capillaries, nylon and nitrocellulose membranes.

[0112] In a sixth aspect, the present invention provides a reagent or a kit, wherein the reagent or the kit comprises the above-mentioned antibody or the above-mentioned antibody conjugate.

[0113] As mentioned above, the antibodies in some embodiments or examples of the present invention can effectively bind to the S100 protein, and therefore, the reagents or kits containing the S100 protein antibodies can effectively detect the S100 protein qualitatively or quantitatively. The reagents or kits provided by the present invention can be used, for example, for detections involving the specific binding properties of the S100 protein and its antibodies, such as immunoblotting and immunoprecipitation. As mentioned above, the antibodies in some embodiments or examples of the present invention have higher binding activity or affinity with the S100 protein, and therefore, the reagents or kits containing the antibodies have higher detection sensitivity or specificity.

[0114] In a seventh aspect, the present invention provides a method for detecting S100 protein, comprising: a) contacting the above-mentioned antibody, antibody conjugate, reagent or kit with S100 protein in a sample to be detected under conditions sufficient for an antibody / antigen binding reaction to form an immune complex; and b) detecting the presence of the immune complex, wherein the presence of the complex indicates the presence of the antigen in the test sample;

[0115] In an alternative embodiment, the immune complex further comprises a second antibody that binds to the antibody.

[0116] In an optional embodiment, the immune complex further comprises a second antibody, and the second antibody binds to the S100 protein.

[0117] In an eighth aspect, the present invention provides use of the above-mentioned anti-S100 protein antibody, antibody conjugate, or the above-mentioned reagent or kit in the preparation of a product for detecting S100 protein.

[0118] In a ninth aspect, the present invention provides a nucleic acid molecule encoding the above-mentioned antibody.

[0119] In a tenth aspect, the present invention provides a vector containing the above-mentioned nucleic acid molecule.

[0120] In an eleventh aspect, the present invention provides a cell containing the above-mentioned vector.

[0121] In a twelfth aspect, the present invention provides a method for preparing an anti-S100 protein antibody, comprising: culturing the cell as described above.

[0122] Based on the amino acid sequence of the anti-S100 protein antibody disclosed in the present invention, those skilled in the art can easily think of using genetic engineering technology or other technologies (chemical synthesis, recombinant expression) to prepare the anti-S100 protein antibody, for example, separating and purifying the antibody from the culture product of a recombinant cell that can recombinantly express the antibody as described in any of the above items, which is easy to achieve for those skilled in the art. Based on this, no matter what technology is used to prepare the anti-S100 protein antibody of the present invention, it belongs to the protection scope of the present invention.

[0123] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0124] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used for the practice or testing of the preparations or unit doses herein, some methods and materials are now described. Unless otherwise stated, the techniques adopted or considered herein are standard methods. Materials, methods and examples are illustrative and non-restrictive only.

[0125] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry, and immunology, which are within the capabilities of a skilled artisan. This technique is fully explained in the literature, such as Molecular Cloning: A Laboratory Manual, 2nd Edition (Sambrook et al., 1989); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Animal Cell Culture (RI Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JM Miller and MP Calos, eds., 1987); Current Protocols in Molecular Biology (FM Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction (PCR: The Polymerase Chain Reaction) (Academic Press, Inc.); Reaction" (Mullis et al., eds., 1994); and Current Protocols in Immunology (JE Coligan et al., eds., 1991), each of which is expressly incorporated herein by reference.

[0126] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0127] Example 1 Preparation of Anti-S100 4G8 Monoclonal Antibody

[0128] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TMThe RACE cDNA Amplification Kit was purchased from Takara. The pMD-18T vector was purchased from Takara. The plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were completed by Invitrogen. The hybridoma cell line secreting the Anti-S100 4G8 monoclonal antibody was prepared in this laboratory and revived for future use.

[0129] (1) Antibody gene preparation

[0130] mRNA was extracted from the hybridoma cell line secreting Anti-S100 4G8 monoclonal antibody, and the DNA product was obtained by RT-PCR. The product was inserted into the pMD-18T vector after A addition reaction with rTaq DNA polymerase and transformed into DH5α competent cells. After the colonies grew, the Heavy Chain and Light Chain gene clones were taken respectively, and 4 clones each were sent to a gene sequencing company for sequencing.

[0131] (2) Sequence analysis of the variable region gene of Anti-S100 4G8 antibody

[0132] The gene sequences obtained by the above sequencing were placed in the kabat antibody database for analysis, and the VNTI11.5 software was used for analysis to determine that the genes amplified by the heavy chain and light chain primer pairs were correct. Among the gene fragments amplified by the Light Chain, the VL gene sequence was 324bp, and there was a 57bp leader peptide sequence in front of it; in the gene fragments amplified by the Heavy Chain primer pair, the VH gene sequence was 366bp, belonging to the VH1 gene family, and there was a 57bp leader peptide sequence in front of it.

[0133] (3) Construction of recombinant antibody expression plasmid

[0134] pcDNA TM 3.4 vector is a recombinant antibody eukaryotic expression vector constructed, which has introduced multiple cloning restriction sites such as HindIII, BamHI, and EcoRI, and is named pcDNA3.4A expression vector, hereinafter referred to as 3.4A expression vector; according to the sequencing results of the antibody variable region genes in the above pMD-18T, the VL and VH gene-specific primers of the antibody were designed, with HindIII, EcoRI restriction sites and protective bases at both ends, respectively, and the 0.70kb Light Chain gene fragment and the 1.40kb Heavy Chain gene fragment were amplified by the PCR amplification method.

[0135] The Heavy Chain and Light Chain gene fragments were double-digested with HindIII / EcoRI, and the 3.4A vector was double-digested with HindIII / EcoRI. After the fragments and vectors were purified and recovered, the Heavy Chain gene and the Light Chain gene were respectively connected to the 3.4A expression vector to obtain the recombinant expression plasmids of Heavy Chain and Light Chain, respectively.

[0136] 2. Recombinant Antibody Production

[0137] Resuscitate HEK293 cells in advance and subculture them to 200 ml system to make the cell density reach 3-5×10 6 cells / ml, the cell density reaches the selected antibody concentration and cells, and the cell viability is >95%; the cells are washed by centrifugation and re-dissolved with culture medium, and the cell density is adjusted to 2.9×10 6 cells / ml, wash the cells, re-dissolve with culture medium, and use it as cell diluent. Prepare plasmid DNA and transfection reagent diluents with culture medium respectively. Add transfection reagent diluent to plasmid DNA diluent, mix well and let stand at room temperature for 15 minutes; slowly add the mixture to cell diluent within 1 minute, mix well and take samples to count, record and observe the viability of cells after transfection, and place them in a 35℃ constant temperature incubator for culture, speed 120rmp, CO2 content 8%, and centrifuge after 13 days. The supernatant of the centrifugation was affinity purified with proteinA affinity chromatography column. Take 6ug layer of purified antibody for reducing SDS-PAGE, and the electrophoresis is shown in the figure. After reducing SDS-PAGE, two bands are shown, one Mr is 50KD (heavy chain) and the other Mr is 28KD (light chain).

[0138] The obtained antibody was named Anti-S100 4G8Rmb1. Anti-S100 4G8Rmb1 was mutated to obtain a mutant antibody. The sequences of the heavy chain (H) and light chain (L) of the above antibody are shown in the following table:

[0139] Table 2: Antibody sequences

[0140] Antibody Name Heavy chain Light chain Anti-S100 4G8Rmb1 SEQ ID NO:34 SEQ ID NO:46 Anti-S100 4G8Rmb2 SEQ ID NO:36 SEQ ID NO:46 Anti-S100 4G8Rmb3 SEQ ID NO:37 SEQ ID NO:46 Anti-S100 4G8Rmb4 SEQ ID NO:40 SEQ ID NO:46 Anti-S100 4G8Rmb5 SEQ ID NO:39 SEQ ID NO:46 Anti-S100 4G8Rmb6 SEQ ID NO:35 SEQ ID NO:46 Anti-S100 4G8Rmb7 SEQ ID NO:38 SEQ ID NO:46 Anti-S100 4G8Rmb8 SEQ ID NO:34 SEQ ID NO:48 Anti-S100 4G8Rmb9 SEQ ID NO:34 SEQ ID NO:50 Anti-S100 4G8Rmb10 SEQ ID NO:34 SEQ ID NO:49 Anti-S100 4G8Rmb11 SEQ ID NO:34 SEQ ID NO:47

[0141] Example 2 Performance testing of antibodies

[0142] 1. Activity identification

[0143] The coating solution (main component NaHCO3) was used to dilute S100 antigen (purchased from Hytest, Cat.#8S9h) to 3ug / ml, 100uL per well, and incubated at 4°C overnight; the next day, the cells were washed twice with washing solution (main component Na2HPO4+NaCl), and patted dry; blocking solution (20% BSA+80% PBS) was added, 120uL per well, and incubated at 37°C for 1h, and patted dry; the diluted purified antibody and Antibody, 100uL / well, 37℃, 30min; wash 5 times with detergent, pat dry; add goat anti-mouse IgG-HRP, 100uL per well, 37℃, 30min; wash 5 times with detergent, pat dry; add colorimetric solution A (50uL / well), add colorimetric solution B (50uL / well), 10min; add stop solution, 50uL / well; read OD value at 450nm (reference 630nm) on the microplate reader.

[0144] Note: Solution A (main ingredients: citric acid + sodium acetate + acetanilide + urea peroxide); Solution B (main ingredients: citric acid + EDTA·2Na + TMB + concentrated HCL); Stop solution (EDTA·2Na + concentrated H2SO4)

[0145] Table 3: Activity data

[0146] Concentration (ng / ml) 500.00 250.00 125.00 62.50 31.25 0.00 Comparison 0.922 0.529 0.312 0.187 0.023 0.005 Anti-S100 4G8Rmb1 1.946 1.755 1.199 0.703 0.434 0.011 Anti-S100 4G8Rmb2 1.090 0.583 0.388 0.288 0.105 0.014 Anti-S100 4G8Rmb3 2.095 1.907 1.670 1.083 0.624 0.006 Anti-S100 4G8Rmb4 1.272 0.644 0.269 0.137 0.102 0.008 Anti-S100 4G8Rmb5 1.953 1.727 1.037 0.526 0.295 0.003 Anti-S100 4G8Rmb6 1.709 1.232 0.699 0.338 0.253 0.004 Anti-S100 4G8Rmb7 1.864 1.250 0.780 0.487 0.223 0.012 Anti-S100 4G8Rmb8 1.861 1.629 0.804 0.480 0.270 0.006 Anti-S100 4G8Rmb9 2.017 1.891 1.077 0.654 0.420 0.000 Anti-S100 4G8Rmb10 2.070 1.438 0.657 0.314 0.241 0.000 Anti-S100 4G8Rmb11 1.686 0.961 0.541 0.266 0.195 0.000

[0147] 2. Stability assessment

[0148] The above antibodies were placed at 4°C (refrigerator), -80°C (refrigerator), and 37°C (constant temperature box) for 21 days, and samples were taken for 7 days, 14 days, and 21 days for status observation, and the activity of the 21-day samples was tested. The results showed that there was no obvious protein state change in the antibodies under the three test conditions for 21 days, and the activity did not show a downward trend with the increase of the test temperature, indicating that the above antibodies are stable. Table 4 below shows the OD results of the enzyme immunoassay activity test of the antibody Anti-S100 4G8Rmb3 for 21 days.

[0149] Table 4: Stability data

[0150] Sample concentration (ng / ml) 125 62.5 0 4℃, 21-day samples 1.627 1.031 0.017 -80℃, 21 days sample 1.633 1.047 0.006 37℃, 21 days sample 1.636 1.039 0.035

[0151] 3. Chemiluminescence platform performance test

[0152] 3.1 Antibody coating process

[0153] 10mg / mL carboxyl magnetic beads, washed 3 times with MES buffer. Resuspend the magnetic beads in MES buffer, add EDC with a final concentration of 1mg / mL, shake at 25℃, and react for 30min. Resuspend the magnetic beads in MES buffer, add antibody Anti-S100-A (from Feifeipeng Bio) solution with a final concentration of 0.2mg / mL, use this solution as magnetic particle working solution, shake at 25℃, and react for 120min. Store in Tris buffer at 2-8℃.

[0154] 3.2 Antibody labeling process

[0155] 3mg / ml of the antibodies Anti-S100 4G8Rmb1~Anti-S100 4G8Rmb11 and the control antibody Anti-S100-B (from Feipeng Bio) were replaced with PBS (100mM PB, 50mM sodium chloride, pH8.0) using a Zeba desalting column (10K MWCO). Acridinium ester was prepared into a 4mM solution using DMSO. 10eq of acridinium ester was added to the Anti-NTProBNP-A antibody solution and reacted at 25°C for 2 hours. The excess reagent was removed by desalting, and the acridinium ester-modified antibody was used as the acridinium ester working solution and stored at 4°C for later use.

[0156] 3.3 Detection process

[0157] Detection was performed on the Yingkai Shine i2910 fully automatic chemiluminescence immunoassay analyzer:

[0158] 1) 50μl S100 antigen quality control + 50μL sample treatment agent + 50μl magnetic bead-labeled antibody (concentration 0.05%), react at 37℃ for 20min; 2) Add 50μl acridinium ester-labeled antibody, react at 37℃ for 20min, wash 3 times with PBST; 3) Add 100μl pre-excitation solution and 100μl excitation solution, detect relative luminescence intensity (RLU), and read the instrument value.

[0159] Remark:

[0160] Samples: Refers to clinical samples containing different concentrations of S100 calibrators and Roche values.

[0161] 3.4 Test results

[0162] 3.4.1 Test results of calibration materials

[0163] The detection results of antibodies Anti-S100 4G8Rmb1 to Anti-S100 4G8Rmb11 on the S100 calibrator are shown in Table 5. The results show that within the concentration range of 2 to 20 ng / ml, antibodies Anti-S100 4G8Rmb1 to Anti-S1004G8Rmb11 can effectively detect.

[0164] Table 5: Chemiluminescence platform calibrator test results

[0165] Calibrator concentration (ng / ml) 0 0.2 2 20 Anti-S100 4G8Rmb1 923 81827 615086 6585527 Anti-S100 4G8Rmb2 684 55517 440239 4182502 Anti-S100 4G8Rmb3 678 82668 629387 5982944 Anti-S100 4G8Rmb4 1133 71811 517713 5320443 Anti-S100 4G8Rmb5 967 72437 544070 5502263 Anti-S100 4G8Rmb6 642 74656 600588 5486459 Anti-S100 4G8Rmb7 1176 90545 690169 6781368 Anti-S100 4G8Rmb8 967 89921 708367 6544050 Anti-S100 4G8Rmb9 639 74408 592781 5297158 Anti-S100 4G8Rmb10 905 75831 603801 5701452 Anti-S100 4G8Rmb11 380 47299 389368 3428602

[0166] 3.4.2 Clinical sample test results

[0167] The test results of clinical samples using antibodies Anti-S100 4G8Rmb1 to Anti-S100 4G8Rmb11 are shown in Table 6. The results show that the clinical correlation R2 of the chemiluminescence reagent composed of antibodies Anti-S100 4G8Rmb1 to Anti-S100 4G8Rmb11 and the Roche S100 reagent is greater than 0.98, which is significantly better than the control antibody.

[0168] Table 6: Clinical sample test results

[0169]

[0170] The amino acid sequences involved in this application are shown in Table 7:

[0171] Table 7: Amino acid sequence

[0172]

[0173]

[0174]

[0175] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An anti-S100 protein antibody, comprising three complementary determining regions of a heavy chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33, and three complementary determining regions of a light chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45.

2. The antibody according to claim 1, characterized in that The complementarity determining regions of the variable regions are defined by any one of the Kabat, Chothia, IMGT, AbM or Contact systems or a combination of multiple systems.

3. An antibody against S100 protein, characterized in that: The antibody comprises the following complementarity determining regions: HCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:1 (NYWIH); HCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:2 (GEINPSNGRTKFNEKFKT); HCDR3 comprising or consisting of the amino acid sequence shown in SEQ ID NO: 3 (RFFIASIGGAMDY), 17 (RFFLASIGGAMDY), 18 (RFFIASLGGAMDY) or 19 (RFFLASLGGAMDY); LCDR1, which comprises or consists of the amino acid sequence shown in SEQ ID NO:4 (SASSSVSSSYLY); LCDR2, which comprises or consists of the amino acid sequence shown in SEQ ID NO:5 (GTSNLAS); and LCDR3, which comprises or consists of the amino acid sequence shown in SEQ ID NO:6 (HQWSSYPPT); Optionally, the antibody further has HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4; Optionally, the HFR1 comprises SEQ ID NO:7 or an amino acid sequence having at least 80% identity thereto; The HFR2 comprises SEQ ID NO: 8 or an amino acid sequence having at least 80% identity thereto; The HFR3 comprises SEQ ID NO:9 or an amino acid sequence having at least 80% identity thereto; The HFR4 comprises SEQ ID NO: 10 or an amino acid sequence having at least 80% identity thereto; The LFR1 comprises SEQ ID NO: 11 or an amino acid sequence having at least 80% identity thereto; The LFR2 comprises SEQ ID NO: 12 or an amino acid sequence having at least 80% identity thereto; The LFR3 comprises SEQ ID NO: 13 or an amino acid sequence having at least 80% identity thereto; and The LFR4 comprises SEQ ID NO: 14 or an amino acid sequence having at least 80% identity thereto.

4. An anti-S100 protein antibody comprising a heavy chain variable region and / or a light chain variable region, characterized in that: The amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NOs: 27, 28, 29, 30, 31, 32, and 33; the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NOs: 41, 42, 43, 44, and 45; Optionally, the combination of the heavy chain variable region and the light chain variable region is selected from any one of the following combinations: Optionally, the antibody further comprises a constant region; Optionally, the constant region includes a heavy chain constant region and / or a light chain constant region; Optionally, the heavy chain constant region is selected from any one of the heavy chain constant regions of IgG, IgA, IgM, IgE, and IgD, or a combination of multiple constant region segments; Optionally, the heavy chain constant region includes CH1 of IgG, hinge region of IgG, CH2 of IgM, CH3 of IgM and / or CH4 of IgM; Optionally, the species of the constant region is cattle, horse, pig, sheep, goat, rat, mouse, dog, camel, cat, rabbit, donkey, deer, mink, chicken, duck, goose or human; Optionally, the species origin of the constant region is mouse; Optionally, the heavy chain constant region sequence is as shown in SEQ ID NO: 15 or has at least 80% identity thereto; Optionally, the light chain constant region sequence is as shown in SEQ ID NO: 16 or has at least 80% identity thereto.

5. An antibody against S100 protein, comprising a heavy chain and / or a light chain, characterized in that: The amino acid sequence of the heavy chain is shown in any one of SEQ ID NOs: 34, 35, 36, 37, 38, 39, and 40; the amino acid sequence of the light chain is shown in any one of SEQ ID NOs: 46, 47, 48, 49, and 50.

6. An antibody conjugate, characterized in that: The antibody conjugate comprises the antibody according to any one of claims 1 to 5; Optionally, the antibody conjugate further comprises biotin or a biotin derivative conjugated to the antibody; Optionally, the antibody conjugate further comprises a label coupled to the antibody; Optionally, the label is selected from fluorescent dyes, enzymes, radioisotopes, chemiluminescent agents and nanoparticle labels; Optionally, the antibody conjugate further comprises a solid phase carrier coupled to the antibody.

7. A reagent or a kit, characterized in that: The reagent or kit comprises the antibody according to any one of claims 1 to 5 or the antibody conjugate according to claim 6.

8. Use of the antibody according to any one of claims 1 to 5, the antibody conjugate according to claim 6, or the reagent or kit according to claim 7 in the preparation of a product for detecting S100 protein; Optionally, the use includes: a) contacting the antibody according to any one of claims 1 to 5, the antibody conjugate according to claim 6, or the reagent or kit according to claim 7 with the S100 protein in a sample to be detected under conditions sufficient for an antibody / antigen binding reaction to form an immune complex; and b) detecting the presence of the immune complex, the presence of the complex indicating the presence of the antigen in the test sample; Optionally, the immune complex further comprises a second antibody, which binds to the antibody; Optionally, the immune complex further comprises a second antibody, which binds to the S100 protein.

9. A nucleic acid, a vector, a cell or a method for preparing the antibody according to any one of claims 1 to 5, wherein the nucleic acid encodes the antibody according to any one of claims 1 to 5; the vector contains a nucleic acid encoding the antibody according to any one of claims 1 to 5; the cell contains the above nucleic acid or vector; the method comprises the above cell.

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