Anti-TAT monoclonal antibody as well as coding sequence and application thereof

Abstract

The invention discloses an anti-TAT monoclonal antibody and a coding sequence and application thereof, the antibody or an antigen binding fragment of the antibody can be specifically bound with TAT, and the antibody or the antigen binding fragment of the antibody comprises heavy chain variable regions CDR-H1, CDRH2 and CDR-H3 and light chain variable regions CDR-L1, CDR-L2 and CDR-L3. The antibody or the antigen binding fragment thereof can be specifically bound with TAA and used for preparing a chemiluminescent reagent and quantitatively detecting TAT, the detection sensitivity of 0.4 ng / mL is achieved, the correlation coefficient R2 is larger than or equal to 0.950, the antibody or the antigen binding fragment thereof has high correlation with a measured value of a clinical plasma sample, and the requirement for clinical detection is met.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine or biopharmaceutical technology, and in particular to an anti-TAT monoclonal antibody, its encoding sequence, and its applications. Background Technology

[0002] Thrombin-antithin III complex (TAT) is an important substance produced during blood clotting. Its production involves a series of complex physiological processes. When blood vessels are damaged, the body's coagulation system is activated. Prothrombin is activated and converted into thrombin under the action of various coagulation factors. Thrombin is a key coagulation factor that catalyzes the conversion of fibrinogen into fibrin, which then forms a thrombus to stop bleeding. Antithrombin III (ATIII) is an important natural anticoagulant protein in the human body that can specifically bind to thrombin to form the thrombin-antithin III complex (TAT).

[0003] Changes in TAT levels play a crucial indicative role. In thrombotic diseases such as deep vein thrombosis (DVT) and pulmonary embolism, plasma TAT levels are often significantly elevated. Taking DVT as an example, studies have found that TAT levels can rise rapidly in the early stages of the disease, with some patients experiencing plasma TAT concentrations several times or even higher than normal. This is because during thrombosis, the coagulation system is overactivated, producing large amounts of thrombin, which binds to antithrombin III to generate even more TAT. By monitoring TAT levels, doctors can assist in diagnosing these diseases and assess the severity of the condition and the effectiveness of treatment based on its changing trends.

[0004] In disseminated intravascular coagulation (DIC), a serious clinical syndrome, TAT levels also change significantly. In DIC, both the systemic coagulation and fibrinolytic systems are disrupted, leading to a large amount of thrombin production and a sharp increase in TAT levels. Furthermore, TAT levels fluctuate continuously as the disease progresses. Dynamic monitoring of TAT levels helps in the timely detection of early signs of DIC and is crucial for guiding clinical treatment, such as the rational use of anticoagulants.

[0005] In addition, TAT levels can be abnormal in some patients with liver disease. The liver is the main site of antithrombin III synthesis; when liver function is impaired, antithrombin III synthesis decreases, and TAT production is also affected. For example, plasma TAT levels in patients with cirrhosis are usually lower than normal. This reflects the important influence of liver function on the coagulation and anticoagulation balance. Measuring TAT can help assess the coagulation function status of patients with liver disease, providing a basis for the prevention and treatment of potential bleeding complications.

[0006] TAT is a molecular marker that reflects the amount and activity of thrombin. Therefore, thrombin levels can be indirectly measured clinically by detecting TAT in the blood. TAT is a molecular marker that measures increased thrombin production and activity. Thrombosis is a pathological process involved in all thrombotic diseases, and excessive activation of the coagulation cascade reaction involving thrombin is one of the factors contributing to thrombosis. Detecting thrombin can reflect the state of the body's coagulation system. However, because thrombin is rapidly eliminated after being produced in the body by reacting with other substances in the blood, it is difficult to detect thrombin levels in the blood.

[0007] TAT has a blood half-life of 3–15 minutes and can be measured. This complex directly reflects the amount of thrombin generated or the degree of activation of the coagulation system, and is an early signal of thrombus formation. Its clinical significance includes: identifying the initiation of the coagulation system, serving as an early marker of coagulation system activation, and indicating thrombin generation; persistently elevated TAT levels indicate that the patient is in a hypercoagulable state, with a significantly increased risk of thrombosis, serving as an early signal of thrombus formation; a sensitive indicator to help determine the optimal timing of clinical anticoagulation therapy; and monitoring the efficacy of thrombolysis, especially the rethrombosis monitoring after thrombolytic therapy.

[0008] During thrombolytic therapy, a decrease in TAT levels indicates effective treatment; a subsequent increase suggests an increased risk of re-occlusion of the blood vessel. Plasma TAT levels are significantly elevated in disseminated intravascular coagulation (DIC), deep vein thrombosis (DVT), or pulmonary embolism, and a persistently elevated level indicates a poor prognosis.

[0009] TAT detects thrombosis risk earlier than traditional coagulation markers (such as D-dimer) and has high sensitivity. Therefore, the preparation of highly specific and sensitive monoclonal antibodies for the development of high-quality TAT immunoassay reagents is of great significance in the clinical diagnosis, disease assessment and treatment monitoring of various coagulation-related diseases. Summary of the Invention

[0010] To address one or more problems existing in the prior art, the present invention provides an anti-TAT monoclonal antibody. The technical solution adopted by the present invention to solve the above problems is: an anti-TAT monoclonal antibody, wherein the antibody or its antigen-binding fragment can specifically bind to TAT, and the antibody or its antigen-binding fragment is any one of the following two: The heavy chain variable region of the first antibody or its antigen-binding fragment comprises: CDR-H1 with an amino acid sequence of SEQ ID NO:1 or a variant thereof, CDR-H2 with an amino acid sequence of SEQ ID NO:2 or a variant thereof, and CDR-H3 with an amino acid sequence of SEQ ID NO:3 or a variant thereof; The light chain variable region of the first antibody or its antigen-binding fragment comprises: CDR-L1 with an amino acid sequence of SEQ ID NO:4 or a variant thereof, CDR-L2 with an amino acid sequence of SEQ ID NO:5 or a variant thereof, and CDR-L3 with an amino acid sequence of SEQ ID NO:6 or a variant thereof; The heavy chain variable region of the second type of antibody or its antigen-binding fragment comprises: CDR-H1 with an amino acid sequence of SEQ ID NO:7 or a variant thereof, CDR-H2 with an amino acid sequence of SEQ ID NO:8 or a variant thereof, and CDR-H3 with an amino acid sequence of SEQ ID NO:9 or a variant thereof; The second type of antibody or its antigen-binding fragment comprises a light chain variable region containing: CDR-L1 with an amino acid sequence of SEQ ID NO:10 or a variant thereof, CDR-L2 with an amino acid sequence of SEQ ID NO:11 or a variant thereof, and CDR-L3 with an amino acid sequence of SEQ ID NO:12 or a variant thereof.

[0011] In some embodiments, the variant of the amino acid sequence contains an amino acid mutation compared to its source amino acid sequence, the amino acid mutation being any one or more of the substitution, deletion, or addition of one or more amino acids.

[0012] Furthermore, the substitutions in the amino acid mutations are conservative substitutions.

[0013] Furthermore, the identity of the variant of the amino acid sequence to its derived amino acid sequence is any one of at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, and 100%.

[0014] In some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain constant region and a light chain constant region, wherein the constant region is a heavy chain constant region and a light chain constant region of any of the following species: mouse, rat, or human.

[0015] Furthermore, the heavy chain constant region is the IgG heavy chain constant region, and the light chain constant region is the κ or λ light chain constant region.

[0016] In some embodiments, the antibody or its antigen-binding fragment is any one of IgG, IgM, IgE, IgD, and IgA antibodies.

[0017] In some embodiments, the antigen-binding fragment is selected from any one of scFv, Fab, (Fab')2, Fd, Fv, CDR fragment, nanobody, disulfide-linked Fv(dsFv), biantibody, bispecific antibody, and multispecific antibody; and / or, the antibody is any one of murine antibody, chimeric antibody, and humanized antibody.

[0018] Application of an anti-TAT monoclonal antibody, wherein the antibody or its antigen-binding fragment described above is applied to any one or more of chemiluminescent reagent detection, quantitative detection of TAT, and immunological detection.

[0019] A nucleic acid sequence for encoding the antibody or its antigen-binding fragment described above, wherein the nucleic acid sequence SEQ ID NO:13-24 sequentially encodes the amino acid sequences SEQ ID NO:1-12.

[0020] The technical advantages achieved by this invention are: the antibody or its antigen-binding fragment provided in this application can specifically bind to TAA and be used to prepare chemiluminescent reagents and quantitatively detect TAT, achieving a detection sensitivity of 0.4 ng / mL and a correlation coefficient R. 2 ≥ 0.950, which shows a high correlation with the measured value of clinical plasma samples and meets the requirements of clinical testing. Attached Figure Description

[0021] Figure 1 Example 1 - Reagent Correlation Test: Correlation analysis between reagents prepared using combination YM0243 / AT3-19-4E4 and clinical measurements; Figure 2 Example - Reagent Correlation Test: Correlation analysis between reagents prepared by combination YM0243 / AT3-19-4C5 and clinical measurements. Detailed Implementation

[0022] To make the above-mentioned objects, features, and advantages of the present invention more readily understood, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0023] This invention discloses an anti-TAT monoclonal antibody, wherein the antibody or its antigen-binding fragment can specifically bind to TAT, and the antibody or its antigen-binding fragment is any one of the following two (the amino acid and nucleic acid sequences of the TAT antibody CDR region in Table 1). The heavy chain variable region of the first antibody or its antigen-binding fragment comprises: CDR-H1 with an amino acid sequence of SEQ ID NO:1 or a variant thereof, CDR-H2 with an amino acid sequence of SEQ ID NO:2 or a variant thereof, and CDR-H3 with an amino acid sequence of SEQ ID NO:3 or a variant thereof; The light chain variable region of the first antibody or its antigen-binding fragment comprises: CDR-L1 with an amino acid sequence of SEQ ID NO:4 or a variant thereof, CDR-L2 with an amino acid sequence of SEQ ID NO:5 or a variant thereof, and CDR-L3 with an amino acid sequence of SEQ ID NO:6 or a variant thereof; The heavy chain variable region of the second type of antibody or its antigen-binding fragment comprises: CDR-H1 with an amino acid sequence of SEQ ID NO:7 or a variant thereof, CDR-H2 with an amino acid sequence of SEQ ID NO:8 or a variant thereof, and CDR-H3 with an amino acid sequence of SEQ ID NO:9 or a variant thereof; The light chain variable region of the second type of antibody or its antigen-binding fragment comprises: CDR-L1 with an amino acid sequence of SEQ ID NO:10 or a variant thereof, CDR-L2 with an amino acid sequence of SEQ ID NO:11 or a variant thereof, and CDR-L3 with an amino acid sequence of SEQ ID NO:12 or a variant thereof. Each of the above includes a heavy chain variable region and a light chain variable region, wherein both the heavy chain variable region and the light chain variable region contain three CDRs and are defined by the Kabat numbering system.

[0024] A nucleic acid sequence encoding the antibody described above or its antigen-binding fragment, as shown in Table 1 for the amino acid and nucleic acid sequences of the TAT antibody CDR region, wherein the nucleic acid sequence SEQ ID NO:13-24 sequentially encodes the amino acid sequences SEQ ID NO:1-12.

[0025] Application of an anti-TAT monoclonal antibody, wherein the antibody or its antigen-binding fragment is used in any one or more of chemiluminescent reagent detection, quantitative detection of TAT, and immunological detection. "Immunological detection" refers to assays that utilize the specific interaction / binding between antigen and antibody, and are generally used to detect the presence or level of a specific antigen or antibody in a sample. Such immunological assays are well known to those skilled in the art and include, but are not limited to, enzyme immunoassay (EIA), chemiluminescent immunoassay (CLIA), radioimmunoassay (RIA), fluorescence immunoassay (FIA), Western blotting, immunoturbidimetry, and surface plasmon resonance assays.

[0026] It should be noted that the variant of the amino acid sequence contains amino acid mutations compared to its source amino acid sequence. These mutations are any one or more substitutions, deletions, or additions of one or more amino acids (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, or 5 amino acids). Preferably, the substitutions in the amino acid mutations are conserved substitutions. The identity of the variant of the amino acid sequence to its source amino acid sequence is any one of at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%.

[0027] Specifically, in some embodiments, the antibody or its antigen-binding fragment comprises a heavy chain constant region (CH) and a light chain constant region (CL), wherein the constant region is a heavy chain constant region and a light chain constant region of any species of mouse, rat, or human; wherein the heavy chain constant region is an IgG heavy chain constant region, such as the IgG1, IgG2, IgG3, or IgG4 heavy chain constant region, or for example, the mouse IgG1, IgG2a, IgG2b, or IgG3 constant region; and the light chain constant region is a κ or λ light chain constant region, such as the human λ light chain constant region. In some embodiments, the antibody or its antigen-binding fragment is any one of IgG, IgM, IgE, IgD, or IgA antibodies.

[0028] In some embodiments, the antigen-binding fragment is selected from any one of scFv, Fab, (Fab')2, Fd, Fv, CDR fragment, nanobody, disulfide-linked Fv (dsFv), diabody, bispecific antibody, and multispecific antibody; and / or, the antibody is any one of murine antibody, chimeric antibody, and humanized antibody.

[0029] The term "complementarity-determining region" or "CDR" refers to the amino acid residues in the variable region of an antibody responsible for antigen binding. Each of the heavy and light chain variable regions contains three CDRs, designated CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, such as the Kabat numbering system. For a given antibody, those skilled in the art will readily identify the CDRs as defined by each numbering system. In this invention, the CDRs contained in the antibody or its antigen-binding fragment can be determined according to various numbering systems known in the art.

[0030] As used in this application, the term "framework region" or "FR" residue refers to those amino acid residues in the antibody variable region other than the CDR residues as defined above. The term "antibody" is not limited to any particular method of antibody production. For example, it includes recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. Antibodies can be different isotypes of antibodies, such as IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies.

[0031] Table 1: Amino acid and nucleic acid sequences of the CDR region of TAT antibody Example 1 - Preparation of anti-TAT monoclonal antibody: 1. Animal immunization, synthesize the gene sequence of antithrombin III (sequence 1) and insert it into the expression vector pCAG (with BamHI / EcoRI restriction sites on both sides), construct and extract the expression vector pCAG-ATIII; Babl / c mice were immunized with the extracted expression vector at a dose of 60-100 μg / mouse. A second immunization was performed two weeks later. Three days after the fourth immunization, tail vein blood was collected to test serum titers. Cell fusion was initiated when the titer was greater than 400,000. Three days prior to cell fusion, intraperitoneal pulse immunization was performed using TAT complex antigen (catalog number YA0212, Shenzhen Yawei Century Technology Co., Ltd.) at a dose of 60-100 μg / mouse. Sequence 1: MYSNVIGTVTSGKRKVYLLSLLLIGFWDCVTCHGSPVDICTAKPRDIPMNPMCIYRSPEKKATEDEGSEQKIPEATNRRVWELSKANSRFATTFYQHLADSKNDNDNIFLSPLSIS TAFAMTKLGACNDTLQQLMEVFKFDTISEKTSDQIHFFFAKLNCRLYRKANKSSKLVSANRLFGDKSLTFNETYQDISELVYGAKLQPLDFKENAEQSRAAINKWVSNKTEGRITD VIPSEAINELTVLVLVNTIYFKGLWKSKFSPENTRKELFYKADGESCSASMMYQEGKFRYRRVAEGTQVLELPFKGDDITMVLILPKPEKSLAKVEKELTPEVLQEWLDELEEMML VVHMPRFRIEDGFSLKEQLQDMGLVDLFSPEKSKLPGIVAEGRDDLYVSDAFHKAFLEVNEEGSEAAASTAVVIAGRSLNPNRVTFKANRPFLVFIREVPLNTIIFMGRVANPCVK 2. SP2 / 0 cell treatment: Mouse myeloma cells (SP2 / 0 cells) were cultured for one week, then digested with trypsin, collected, centrifuged and the supernatant was discarded. The SP2 / 0 cells were washed three times with 30 ml LDMEM basal medium, and finally resuspended in 20 ml LDMEM basal medium.

[0032] 3. Spleen cell isolation and shock immunization: The spleen of the above-mentioned experimental mice was taken, ground into a homogenate, and 30 ml of 1XPBS (buffered saline) was added. Red blood cells were lysed using methods known in the art, and then resuspended in culture medium to obtain isolated spleen cells.

[0033] 4. Cell fusion: Mix spleen cells and SP2 / 0 cells at a ratio of 2:1, centrifuge to remove the supernatant, and slowly add PEG to mix; add 14 mL of DMEM basal medium and incubate at 37°C; add 20 mL of DMEM basal medium, centrifuge to remove the supernatant, and plate the cells with DMEM medium containing HAT. Culture the cells for 6-10 days. In this example, the DMEM basal medium was purchased from Hyclone.

[0034] 5. Select fusion cells, plate them with TAT complex antigen (catalog number YA0212, Shenzhen Yawei Century Technology Co., Ltd.), screen positive clones by indirect ELISA, and prepare single clones by limiting dilution method and ELISA method; obtain cell lines AT3-19-4E4 and AT3-19-4C5 that can specifically bind to TAT complex and antithrombin III at the same time.

[0035] 6. Antibody preparation: Paraffin was injected into the peritoneal cavity of mice, and then the hybridoma cells obtained above were injected into the peritoneal cavity of mice. After the mouse abdomen swelled, the ascites fluid was collected. The mouse ascites fluid was centrifuged at 4°C, the supernatant was collected, filtered through a 0.45µm filter membrane, and air bubbles in the solution were removed by sonication. The purified antibody was obtained by elution with 50mM glycine solution using the Protein G pre-packed column purification method.

[0036] 7. Sequence acquisition: Total RNA was extracted from the monoclonal hybridoma cells obtained in this example using a commercially available RNA Isolater Total RNA Extraction Reagent kit. The total RNA was reverse transcribed, amplified by PCR, and the amplified products were sequenced to obtain the amino acid sequence of the antibody produced by the hybridoma cells and the corresponding DNA coding sequence.

[0037] Example 2 - Preparation of chemiluminescent reagents: 1. Preparation of magnetic bead reagent: Purchased monoclonal antibody YM0243 (Shenzhen Yawei Century Technology Co., Ltd.) was conjugated with toluenesulfonyl magnetic beads (manufacturer: JSR or Thermo). The conjugation reaction buffer was CBS, pH 9.6, and the conjugation catalyst was a conjugation buffer solution containing 3M ammonium sulfate. The magnetic bead conjugation procedure was as follows: 20 μg antibody / mg magnetic bead, magnetic bead reaction volume was 10 mg / ml, reaction buffer consisted of 2 / 3 conjugation reaction buffer and 1 / 3 conjugation catalyst, reaction was carried out at 37℃ for 16-24 h, excess unconjugated magnetic beads were washed with PBST, and excess unconjugated magnetic beads were blocked with PBS containing 5% casein and 5% BSA, pH 7.4, for 12-24 h. After the blocking process is complete, add 50 mM TBS, 5% BSA, 0.02% Tween 20, and 0.05% Proclin 300 to dilute the magnetic beads to a concentration of 0.15-0.3 mg / ml, thus obtaining the working solution of reagent 1 magnetic beads.

[0038] 2. Preparation of luminescent labeling reagent: Take 1 mg of AT3-19-4E4 or AT3-19-4C5 antibody and add 30 μg of NHS-SA-acridine. Incubate at room temperature for 2 h. After purification by molecular sieve, obtain the labeled antibody solution. Dilute with acridine diluent to 0.1-0.4 μg / ml according to the antibody concentration. The acridine diluent is 50 mM MES, 0.15 M NaCl, 0.5% BSA, 0.05% Tween 20, and 0.5% casein to obtain the acridine working solution.

[0039] 3. Luminescent substrate reagents: Luminescent reagent 1: 0.1 mol / L hydrochloric acid + 0.3% hydrogen peroxide; Luminescent reagent 2: 0.15 mol / L sodium hydroxide + 0.05% Triton 100.

[0040] 4. Reagent kit reaction procedure: 50 μL sample, 50 μL magnetic bead working solution, 50 μL acridine working solution, incubate for 20 min, magnetically separate and wash 3 times, add 1-100 μL luminescent reagent, read the luminescence signal value after adding 2-100 μL luminescent reagent; the entire reaction system can be completed in 25 min.

[0041] Example 3 - Reagent Sensitivity Test: Test method: Five samples with concentrations close to the detection limit of 0.4 ng / mL were tested, with each sample tested five times. Test standard: The number of test results below the blank limit of 0.2 ng / mL should be less than or equal to three.

[0042] The test results of the reagent prepared by combination YM0243 / AT3-19-4E4 are shown in Table 2, and the test results of the reagent prepared by combination YM0243 / AT3-19-4C5 are shown in Table 3. It can be seen that the detection sensitivity of both antibody combinations reached 0.4 ng / mL.

[0043] Table 2: Sensitivity Test Results of YM0243 / AT3-19-4E4 Combination Table 3: Sensitivity Test Results of YM0243 / AT3-19-4C5 Combination Example 4 - Reagent Correlation Test: Forty to fifty clinical plasma samples were selected and tested using a self-prepared luminescent reagent. Correlation analysis was performed on the test results, and the results are shown in Table 4.

[0044] Table 4: Antibody-pair correlation detection Correlation analysis between the reagent preparation of combination YM0243 / AT3-19-4E4 and clinical measurements (detection method: Sysmex TAT chemiluminescence reagent kit) is shown in [link to relevant documentation]. Figure 1 Correlation coefficient R 2 = 0.9771; Correlation analysis between the reagents prepared by combination YM0243 / AT3-19-4C5 and clinical measurements is shown in [link to relevant data]. Figure 2 Correlation coefficient R 2 =0.9856. The reagents prepared from both antibody combinations showed good correlation with clinical measurements.

[0045] In summary, the antibody or its antigen-binding fragment provided in this application can specifically bind to TAA and be used to prepare chemiluminescent reagents and quantitatively detect TAT, achieving a detection sensitivity of 0.4 ng / mL and a correlation coefficient R. 2 ≥ 0.950, which shows a high correlation with the measured value of clinical plasma samples and meets the requirements of clinical testing.

[0046] The embodiments described above are merely illustrative of one or more implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the appended claims.

Claims

1. An anti-TAT monoclonal antibody, characterized in that, The antibody or its antigen-binding fragment can specifically bind to TAT, and the antibody or its antigen-binding fragment is any one of the following two: The heavy chain variable region of the first antibody or its antigen-binding fragment comprises: heavy chain variable region CDR1-3, the amino acid sequences of which are shown in SEQ ID NO:1-3 respectively; The light chain variable region of the first antibody or its antigen-binding fragment comprises: light chain variable regions CDR1-3, whose amino acid sequences are shown in SEQ ID NO:4-6 respectively; The second type of antibody or its antigen-binding fragment comprises a heavy chain variable region CDR1-3, the amino acid sequences of which are shown in SEQ ID NO:7-9 respectively; The second type of antibody or its antigen-binding fragment comprises a light chain variable region CDR1-3, the amino acid sequences of which are shown in SEQ ID NO:10-12 respectively. Among them, SEQ ID NO:1: GYSFTDYN, SEQ ID NO:2: IDPYNGGT, SEQ ID NO:3: AAYSDTYHDY, SEQ ID NO:4: SSVSSSY, SEQ ID NO:5: STS, SEQ ID NO:6: QQYSDYPLT, SEQ ID NO:7: GYSFTDYN, SEQ ID NO:8: IDLYNGGS, SEQ ID NO:9: AAYSSTYHDY, SEQ ID NO:10: SSVSY, SEQ ID NO:11: TTS, SEQ ID NO:12: QQRSSYPPT.

2. The anti-TAT monoclonal antibody according to claim 1, characterized in that, The antibody or its antigen-binding fragment comprises a heavy chain constant region and a light chain constant region, wherein the constant region is the heavy chain constant region and light chain constant region of any of the following species: mouse, rat, or human.

3. The anti-TAT monoclonal antibody according to claim 2, characterized in that, The heavy chain constant region is the IgG heavy chain constant region, and the light chain constant region is the κ or λ light chain constant region.

4. The anti-TAT monoclonal antibody according to claim 1, characterized in that, The antibody or its antigen-binding fragment is any one of IgG, IgM, IgE, IgD, or IgA antibodies.

5. The anti-TAT monoclonal antibody according to claim 1, characterized in that, The antigen-binding fragment is selected from any one of scFv, Fab, (Fab')2, Fd, Fv, CDR fragment, nanobody, disulfide-linked Fv (dsFv), biantibody, bispecific antibody, and multispecific antibody; And / or, the antibody is any one of mouse antibody, chimeric antibody, or humanized antibody.

6. The application of an anti-TAT monoclonal antibody, characterized in that, The antibody or its antigen-binding fragment as described in claim 1 may be applied to any one or more of chemiluminescent reagent detection, quantitative detection of TAT, and immunological detection.

7. A nucleic acid sequence, characterized in that, The nucleic acid sequence is used to encode the antibody or its antigen-binding fragment as described in claim 1, wherein the nucleic acid sequence SEQ ID NO:13-24 sequentially encodes the amino acid sequence SEQ ID NO:1-12; Among them, SEQ ID NO:13: GGTTACTCATTCACTGACTACAAC, SEQ ID NO:14: ATTGATCCTTACAATGGTGGTACT, SEQ ID NO:15: GCAGCCTACTCTGATACCTACCATGACTAC, SEQ ID NO:16: TCAAGTGTAAGTTCCAGTTAC, SEQ ID NO:17: AGCACATCC, SEQ ID NO:18: CAGCAGTACAGTGATTACCCACTCACG, SEQ ID NO:19: AGGCCACATTGACTGTTGACAAGT, SEQ ID NO:20: ATTGATCTTTACAATGGTGGTTCT, SEQ ID NO:21: GCAGCCTACTCTAGTACCTACCATGACTAC, SEQ ID NO:22: TCAAGTGTAAGTTAC, SEQ ID NO:23: ACCACATCC, SEQ ID NO:24: CAGCAAAGGAGTAGTTACCCACCAACG.

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