Monoclonal antibody of anti-human PSP protein and application thereof

Abstract

The invention belongs to the technical field of immunodetection, and discloses an anti-human PSP protein monoclonal antibody and application thereof. The monoclonal antibody for resisting the human PSP protein can be combined with the human PSP protein in a high-specificity mode and has high affinity. The anti-human PSP monoclonal antibody provided by the invention can also be prepared into various immunodetection kits for detecting human PSP protein, and especially can be applied to the immunodetection kits prepared by a double-antibody sandwich ELISA (Enzyme-Linked Immunosorbent Assay) or chemiluminescence method, a coating antibody is an antibody mAb21, a detection antibody is an antibody mAb22, the detection sensitivity is less than 15 pg / ml, and the anti-human PSP monoclonal antibody has very high clinical application value.

Description

Technical Field

[0001] The present application relates to the field of immunoassay technology, and in particular to anti-human PSP monoclonal antibodies and applications thereof. Background Art

[0002] PSP protein, whose full name is Phosphoserine Phosphatase or Pancreatic Stone Protein, is a protein with multiple functions in organisms.

[0003] Depending on the source and preparation method, the molecular weight of PSP protein is generally between 25-30kDa. PSP protein is mainly produced by the pancreas and intestines, and can also be expressed in other tissues such as the liver. In the human body, PSP protein is widely distributed in a variety of tissues and organs, playing an important physiological function. PSP has a catalytic effect. PSP protein has the activity of phosphoserine phosphatase, which can catalyze the dephosphorylation of O-phospho-L-serine to L-serine, which is the last step in the biosynthesis of L-serine. L-serine is a precursor for protein synthesis, other amino acid production, nucleotide metabolism and glutathione synthesis, so PSP protein plays an important role in maintaining cell metabolism and physiological function. PSP can also inhibit crystal growth. In the pancreas, PSP protein can inhibit the growth and nucleation of calcium carbonate crystals in pancreatic juice, thereby preventing the formation of pancreatic stones.

[0004] According to research, PSP protein has been shown to be a sensitive and specific biomarker for sepsis. In patients with sepsis, the level of PSP protein is significantly increased, and its increase precedes the onset of clinical symptoms. Compared with traditional biomarkers such as C-reactive protein (CRP) and procalcitonin (PCT), PSP protein has higher accuracy and specificity in diagnosing sepsis. PSP protein can also be used to predict the severity and prognosis of patients with sepsis and to guide treatment decisions. In addition to sepsis, the level of PSP protein may also change in other diseases such as acute pancreatitis, pancreatic cancer, gastric cancer, etc. Therefore, PSP protein may have a wider range of clinical application value.

[0005] At present, the detection methods of PSP protein mainly include enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, etc. These methods have the advantages of high sensitivity and strong specificity. Therefore, it is extremely important to have PSP protein antibodies with good specificity and high affinity. Summary of the invention

[0006] In order to overcome the above technical problems, the present application provides anti-human PSP monoclonal antibodies with good specificity and high affinity, as well as hybridoma cell lines capable of secreting anti-human PSP monoclonal antibodies, and also provides a kit containing anti-human PSP monoclonal antibodies, providing a new approach for the clinical detection of sepsis.

[0007] In one aspect, the present application provides an antibody or antigen-binding fragment against human PSP protein, wherein the antibody or antigen-binding fragment comprises: 1) an antibody or antigen-binding fragment mAb21, wherein the antibody or antigen-binding fragment mAb21 comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, wherein the amino acid sequences of the LCDR1-3 sequentially comprise SSVSY (SEQ ID NO: 1), DTS (SEQ ID NO: 2), and QQWSSNPPT (SEQ ID NO: 3), and the heavy chain variable region comprises HCDR1-3, wherein the amino acid sequences of the HCDR1-3 sequentially comprise GYTFSTYW (SEQ ID NO: 4), IFPGSGST (SEQ ID NO: 5), and VRKRYPYYFDY (SEQ ID NO: 6); and / or, 2) an antibody or antigen-binding fragment mAb22, wherein the antibody or antigen-binding fragment mAb22 comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, wherein the amino acid sequences of the LCDR1-3 sequentially comprise QSMSND (SEQ ID NO: 7), YAS (SEQ ID NO: 8), and QQDYSSPLT (SEQ ID NO: 9), and the heavy chain variable region comprises HCDR1-3, wherein the amino acid sequences of the HCDR1-3 sequentially comprise GYTFTEYT (SEQ ID NO: 10), INPNNGGT (SEQ ID NO: 11), and ARYDFDVLFDV (SEQ ID NO: 12).

[0008] Furthermore, the light chain variable region of the antibody or antigen-binding fragment mAb21 comprises the amino acid sequence shown in SEQ ID NO:13, and the heavy chain variable region of the antibody or antigen-binding fragment mAb21 comprises the amino acid sequence shown in SEQ ID NO:14.

[0009] Furthermore, the light chain variable region of the antibody or antigen-binding fragment mAb22 comprises the amino acid sequence shown in SEQ ID NO:15, and the heavy chain variable region of the antibody or antigen-binding fragment mAb22 comprises the amino acid sequence shown in SEQ ID NO:16.

[0010] On the other hand, the present application also provides a hybridoma cell line mAb21 that secretes anti-human PSP protein monoclonal antibodies, and its preservation information is as follows: taxonomic name: anti-human PSP monoclonal hybridoma cell line, full name of the preservation unit: General Microbiology Center of China Microbiological Culture Collection Administration, abbreviation of the preservation unit: CGMCC, address of the preservation unit: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, preservation date: November 20, 2024, and preservation number: CGMCC No.46136.

[0011] On the other hand, the present application also provides a hybridoma cell line mAb22 that secretes anti-human PSP protein monoclonal antibodies, and its preservation information is as follows: taxonomic name: anti-human PSP monoclonal hybridoma cell line, full name of the preservation unit: General Microbiology Center of China Microbiological Culture Collection Administration, abbreviation of the preservation unit: CGMCC, address of the preservation unit: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, preservation date: November 20, 2024, and preservation number: CGMCC No.46137.

[0012] On the other hand, the present application also provides a monoclonal antibody against human PSP protein, wherein the monoclonal antibody against human PSP protein is obtained by secretion of the hybridoma cell line mAb21 and / or is obtained by secretion of the hybridoma cell line mAb22.

[0013] On the other hand, the present application also provides a kit for detecting human PSP protein, the kit comprising a coating antibody and a detection antibody, the coating antibody is antibody mAb21, and the detection antibody is antibody mAb22.

[0014] Furthermore, the kit is a double antibody sandwich ELISA detection kit or an immunoassay kit prepared by chemiluminescence method.

[0015] On the other hand, the present application also provides the use of the antibody or antigen-binding fragment or the monoclonal antibody in detecting human PSP protein or preparing a kit for detecting human PSP protein.

[0016] On the other hand, the present application also provides the use of the hybridoma cell line in preparing monoclonal antibodies for detecting human PSP protein.

[0017] Compared with the prior art, the monoclonal antibody against human PSP protein of the present application can bind to human PSP protein with high specificity and high affinity. The monoclonal antibody against human PSP of the present application can also be prepared into various immunoassay kits for detecting human PSP protein, especially, applied to immunoassay kits prepared by double antibody sandwich ELISA or chemiluminescence method, wherein the coating antibody is antibody mAb21, and the detection antibody is antibody mAb22, and the detection sensitivity is less than 15pg / ml, which has high clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The double antibody sandwich ELISA method is used to detect the human PSP standard antigen. The horizontal axis is the human PSP standard antigen concentration (pg / ml), and the vertical axis is the OD value of the test. 2 =0.9956, linear detection range 0-1000pg / mL, the sample concentration calculation formula is: y=0.0073x+0.1217. DETAILED DESCRIPTION

[0019] The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions, conditions described in laboratory manuals or conditions recommended by manufacturers.

[0020] Example 1 Preparation of anti-human PSP monoclonal antibody 1) Preparation of human PSP immunogen The PSP gene NM_002909 was selected from Genebank, which encodes a full length of 166 amino acids (aa) of PSP. The immunogen of the antibody of the present invention is PSP 1-160aa. The gene was cloned into the pET23a expression plasmid, with a histag at the C-terminus, expressed in BL21 Escherichia coli, purified by nickel column, and the purity was identified by SDS-PAGE electrophoresis. After electrophoresis, a target protein band with a molecular weight of about 18kDa was observed from the gel, and the purity was above 90%, which met the purity requirements for preparing monoclonal antibodies.

[0021] 2) Animal immunization The purified PSP recombinant protein was emulsified with complete Freund's adjuvant and immunized 6-8 week old BALB / c mice by subcutaneous or intraperitoneal injection, with an immunization dose of 80 μg / mouse. The second immunization was performed two weeks later, emulsified with incomplete Freund's adjuvant, with an immunization dose of 60 μg / mouse. After two immunizations, tail blood was collected and serum titer was determined by gradient dilution using ELISA method; whether to boost immunization was determined based on the titer results, and the mouse with the highest antibody titer was selected for cell fusion.

[0022] 3) Cell fusion and clone screening Take the spleen cells of immune mice and fuse them with mouse myeloma cells SP2 / 0. The spleen cells and SP2 / 0 cells are mixed at a ratio of 10:1, and the fusion is mediated by 50% PEG 4000. After centrifugation, resuspend them in HAT selective culture medium, inoculate them in a 96-well microplate containing feeder cells, and culture them in a 37°C, 5% CO2 incubator. On the 4th and 7th days after fusion, replace the medium with half of the HAT culture medium, and replace it with HT culture medium after 10 days. Observe the growth of hybridoma cells, wait for the clones to grow to 1 / 4 to 1 / 3 of the bottom area of ​​the well, take the culture supernatant, use ELISA method for antibody detection, and screen the positive clone wells. Use the limiting dilution method to clone the hybridoma cells in the positive wells until the antibody positivity rate of the cloned cells is 100%, and select the high secretion specific cell line (that is, the ELISA titer is 1:10 4 The above positive clones) were used to obtain hybridoma cell lines mAb21, mAb22 and mAb23 that secrete anti-human PSP protein monoclonal antibodies.

[0023] 4) Preparation and purification of monoclonal antibodies Hybridoma cell lines mAb21 and mAb22 were cultured in suspension in serum-free medium. When the cell supernatant volume was expanded to the task volume and the cell death rate reached 60%-70% in about one week, the cell suspension was collected, the supernatant was centrifuged, and the antibody was purified by affinity chromatography. The corresponding column material was selected for purification according to the antibody subtype. The concentration of the purified monoclonal antibody was determined by the BCA method, and then the monoclonal antibodies mAb21, mAb22 and mAb23 were obtained.

[0024] Example 2 Identification of anti-human PSP monoclonal antibodies 1) Determination of affinity constant of monoclonal antibodies The affinity constant (Ka) was determined by non-competitive ELISA.

[0025] Coating: Dilute the antigen with carbonate buffer to a concentration of 1, 0.5, 0.1, and 0.05 μg / mL, add 100 μL / well to a 96-well ELISA plate for coating, and incubate at 4°C for 24 h; Blocking: Wash the plate 4 times with PBST, add BSA solution at 200 μL / well, and incubate at 37°C for 2 h; Adding monoclonal antibodies: Wash the plate four times with PBST, dilute the monoclonal antibody OTIR3A11 starting at 100 μg / mL with carbonate buffer, add 100 μL to each well, and incubate at 37°C for 2 h; Add enzyme-labeled secondary antibody: wash the plate 4 times with PBST, add 100 μL of 1:10000-fold diluted HRP enzyme-labeled Ig secondary antibody to each well, and place at 37°C for 30 min; Color development and termination: Wash the plate 4 times with PBST, add 100 μL of substrate color development solution to each well, and react at 37°C in the dark for 15 min; add 50 μL of 1.0 mol / L H2SO4 stop solution to each well to terminate the reaction; Detection: Measure the absorbance value at a wavelength of 450nm (A450nm).

[0026] The S-shaped curve was drawn with the logarithm of antibody concentration as the horizontal axis and the OD value as the vertical axis. After calculation, the affinity constants Ka of anti-human PSP monoclonal antibodies mAb21, mAb22 and mAb23 were all 8×10 7.5 L / mol, 8×10 7.8 L / mol and 8×10 7.3 L / mol.

[0027] 2) Antibody pairing In order to select the best combination of coating antibody and detection antibody, the three purified monoclonal antibodies mAb21, mAb22 and mAb23 were coated on the ELISA plate respectively using the chessboard combination and incubated at 4°C overnight. The next day, the ELISA plate was taken out, washed once with PBST, blocked with 1% BSA solution at 37°C for 2 hours, and washed 3 times with PBST; 100 μl of human PSP full-length protein was added to each well at a concentration of 1 ng / ml, and incubated at 37°C for 1 hour; after the incubation, the ELISA plate was taken out, washed 3 times with PBST, and the above 3 antibodies labeled with HRP were added as detection antibodies, and the 3 monoclonal antibodies and the corresponding enzyme-labeled monoclonal antibodies were paired in pairs for double antibody sandwich ELISA experiments, and incubated at 37°C for 1 hour. Washed 5 times with PBST, TMB substrate was added, and color was developed at 37°C for 10 minutes. After taking out, stop solution was added and OD450 readings were measured on the ELISA instrument. According to the OD value of the sample and the background value of the negative control, the most ideal monoclonal antibody pair was selected, and the pairing screening results are shown in Table 1.

[0028] Table 1 Antibody pairing experiment screening results It can be seen that the antibody mAb21 involved in the present application is the best as a coating antibody, and mAb22 is the best as a detection antibody.

[0029] Example 3 Analysis of the variable region genes and amino acid sequences of monoclonal antibodies mAb21 and mAb22 The recombinant plasmids of mAb21 and mAb22 antibodies were used as DNA templates, and the sequencing primers for the light chain variable region and the heavy chain variable region were designed according to the vector sequences at the 5' ends of the light chain and heavy chain of the templates, and sequencing was performed using a sequencer ABI 3730. The nucleotide sequences of the light chain and heavy chain variable regions of monoclonal antibodies mAb21 and mAb22 were obtained by sequencing.

[0030] The nucleotide sequences of the light chain variable region and the heavy chain variable region were sequenced and analyzed using the IMGT / V-QUEST analysis software on the Internet at http: / / www.imgt.org. The monoclonal antibody mAb21 comprises a light chain variable region and a heavy chain variable region. The light chain variable region comprises LCDR1-3, and the LCDR1-3 respectively comprise the amino acid sequences shown in SEQ ID NOs: 1-3. The heavy chain variable region comprises HCDR1-3, and the HCDR1-3 respectively comprise the amino acid sequences shown in SEQ ID NOs: 4-6.

[0031] The light chain variable region of monoclonal antibody mAb21 comprises the amino acid sequence shown in SEQ ID NO:13, the full length of VL is 107 amino acids, the amino acid numbers of the four domains of FR are 26, 17, 31 and 11, respectively, the amino acid numbers of the three domains of LCDR are 5, 3 and 8, respectively, the regions of LCDR1, LCDR2 and LCDR3 are 27aa-31aa, 49aa-51aa and 88aa-96aa, respectively, and their amino acid sequences are: SSVSY (SEQ ID NO.1), DTS (SEQ ID NO.2), QQWSSNPPT (SEQ ID NO.3).

[0032] The heavy chain variable region of the monoclonal antibody mAb21 comprises the amino acid sequence shown in SEQ ID NO:14, the full length of VH is 115 amino acids, the amino acid numbers of the four domains of its FR are 24, 17, 36 and 11, respectively, the amino acid numbers of the three domains of HCDR are 8, 8 and 11, respectively, HCDR1, HCDR2 and HCDR3 are 25aa-32aa, 50aa-57aa and 94aa-104aa, respectively, and their amino acid sequences are GYTFSTYW (SEQ ID NO.4), IFPGSGST (SEQ ID NO.5), VRKRYPYYFDY (SEQ ID NO.6), respectively.

[0033] The monoclonal antibody mAb22 includes a light chain variable region and a heavy chain variable region, the light chain variable region includes LCDR1-3, and the LCDR1-3 respectively include the amino acid sequences shown in SEQ ID NOs: 7-9, and the heavy chain variable region includes HCDR1-3, and the HCDR1-3 respectively include the amino acid sequences shown in SEQ ID NOs: 10-12.

[0034] The light chain variable region of monoclonal antibody mAb22 comprises the amino acid sequence shown in SEQ ID NO:15, the full length of VL is 107 amino acids, the amino acid numbers of the four domains of FR are 26, 16, 36 and 11, respectively, the amino acid numbers of the three domains of LCDR are 6, 3 and 9, respectively, the regions of LCDR1, LCDR2 and LCDR3 are 27aa-32aa, 49aa-51aa and 88aa-96aa, respectively, and their amino acid sequences are: QSMSND (SEQ ID NO.7), YAS (SEQ ID NO.8), QQDYSSPLT (SEQ ID NO.9), respectively.

[0035] The heavy chain variable region of monoclonal antibody mAb22 comprises the amino acid sequence shown in SEQ ID NO:16, the full length of VH is 115 amino acids, the amino acid numbers of the four domains of FR are 24, 17, 36 and 11, respectively, the amino acid numbers of the three domains of HCDR are 8, 8 and 11, respectively, HCDR1, HCDR2 and HCDR3 are 25aa-32aa, 50aa-57aa and 94aa-104aa, respectively, and their amino acid sequences are GYTFTEYT (SEQ ID NO.10), INPNNGGT (SEQ ID NO.11), and ARYDFDVLFDV (SEQID NO.12), respectively.

[0036] Example 4 Anti-human PSP monoclonal antibodies are used to prepare double antibody sandwich ELISA detection kit Based on the detection technology of ELISA double antibody sandwich method, a human PSP detection kit is prepared for clinical auxiliary diagnosis of diseases related to human PSP.

[0037] 1) Kit composition 1. Strips coated with mAb21 antibody: dilute the antibody to 1 μg / ml with PBS buffer, coat on microtiter plates, 100 μl per well, incubate overnight at 4°C, wash 3 times with PBST, and spin dry; add PBS containing 1% BSA, 5% sucrose, and 0.05% Proclin300 for blocking, react at 37°C for 2h, discard the blocking solution in the wells, and spin dry; place the coated plate in a 37°C oven for 2h to complete the coating, seal it with an aluminum foil bag, and store it at 4°C for future use.

[0038] 2. Reagent Preparation 2.1 Preparation of enzyme conjugates The anti-human PSP detection antibody mAb22 was labeled with a simple sodium periodate method, and the working concentration was titrated to prepare a 1:20 concentrate. The diluent was PBS containing 1% BSA, 5% glycerol, and 0.05% Proclin 300, which was filtered and sterilized.

[0039] 2.2 The washing buffer is conventional PBST at pH 7.4, containing 0.05% Proclin 300, and prepared into a 20-fold concentrated solution.

[0040] 2.3 Enzyme substrate solution (color developing solution): single-component TMB (purchased from the market).

[0041] 2.4 The sample dilution solution contains 1% BSA and 0.05% Proclin 300 in PBS, which is sterilized by filtration.

[0042] 2.5 Stop solution: Add 10 ml of HCl (36-38%) to 110 ml of distilled water and mix slowly to make 1N HCl.

[0043] 2.6 Standard full-length human PSP protein, dilute the sample to 20 μg / ml with PBS containing 1% BSA, 5% sucrose, 10% glycerol, and 0.05% Proclin300, filter sterilize, and aseptically package.

[0044] 2) Key points of detection operation 1. To ensure the accuracy of the test results, it is recommended to set up double-hole determination for both the standard and the sample. A standard curve should be made for each test.

[0045] 2. If the content of the substance to be tested in the specimen is too high, please dilute it with sample diluent first to make the sample meet the detection range of the kit, and then multiply it by the corresponding dilution factor in the final calculation.

[0046] 3. Sample addition: When adding samples, please use disposable clean pipette tips to avoid cross contamination. When adding samples, be as gentle as possible to avoid foaming. Add the sample to the bottom of the ELISA plate well, and do not add along the well wall.

[0047] 4. Incubation: To prevent sample evaporation or contamination, the ELISA plate must be covered with a plate sticker during the incubation process. The ELISA plate should not be in a dry state during the experiment. During the incubation process, the incubator temperature should be observed at any time to see if it is constant at 37°C and adjusted in time. During the incubation process, the incubator should not be opened too many times to avoid affecting the temperature balance.

[0048] 5. Washing: The washing process is very important. Insufficient washing can easily cause false positives.

[0049] 6. Color development: To ensure the accuracy of the experimental results, the stop solution should be added as soon as possible after the substrate reaction time is up. You can observe the color development at regular intervals after adding the substrate solution to control the reaction time (for example, every 10 minutes). When the naked eye can see that the first 3-4 wells of the standard have a clear gradient of blue, and the color development of the last 3-4 wells is not obvious, you can add the stop solution to terminate the reaction. At this time, the blue color immediately turns yellow. The order of adding the stop solution should be as consistent as possible with the order of adding the substrate solution.

[0050] 7. The substrate solution should be light blue or colorless. If the color becomes seriously darker, it must be discarded. The substrate solution is easily contaminated, so please store it properly away from light.

[0051] 3) Determination of standard curves of different dilutions of standard human PSP protein The prepared double antibody sandwich ELISA test kit for detecting human PSP protein was taken out from the 4°C refrigerator and equilibrated to room temperature. The standard was diluted from 15 ng / ml to 250 pg / ml with PBS to prepare 6 series of standard concentrations of 0, 15, 30, 60, 125, and 250. According to the above detection method, 100 μl of the standard was added to each well for incubation, and then the liquid was discarded, and the HRP-labeled detection antibody working solution was added for incubation, and the liquid in the well was discarded, and the substrate solution was added to each well after drying. After color development in the dark, the stop solution was added to each well. Within 5 minutes after the reaction was terminated, the optical density OD value of each well was measured in sequence at a wavelength of 450 nm using an enzyme reader to make a standard curve. The results are shown in Table 2.

[0052] Table 2 Detection of human PSP standards by double antibody sandwich ELISA According to Table 2, the PSP detection standard curve was prepared. Figure 1 The R of the standard curve is shown in 2 =0.9956, linear detection range 0-1000pg / mL, sample concentration calculation formula: y=0.0073x+0.1217, detection sensitivity is less than 15pg / ml.

[0053] 4) Sample testing The above method was used to detect the serum of sepsis patients, and the results are shown in Table 3. According to the results, the mAb21 antibody of the present invention was used as the coating antibody, and the mAb22 antibody was used as the detection antibody. The detection by magnetic microparticles had a good correlation with clinical comparison, and the kit provided an auxiliary means for the diagnosis of diseases related to PSP protein.

[0054] Table 3 Sample testing This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. An antibody or antigen-binding fragment against human PSP protein, characterized in that: The antibody or antigen-binding fragment comprises, 1) an antibody or antigen-binding fragment mAb21, wherein the antibody or antigen-binding fragment mAb21 comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, wherein the amino acid sequences of the LCDR1-3 sequentially comprise SSVSY, DTS, QQWSSNPPT, and the heavy chain variable region comprises HCDR1-3, wherein the amino acid sequences of the HCDR1-3 sequentially comprise GYTFSTYW, IFPGSGST, VRKRYPYYFDY; and / or, 2) The antibody or antigen-binding fragment mAb22, wherein the antibody or antigen-binding fragment mAb22 comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, and the amino acid sequences of the LCDR1-3 comprise QSMSND, YAS, and QQDYSSPLT in sequence, and the heavy chain variable region comprises HCDR1-3, and the amino acid sequences of the HCDR1-3 comprise GYTFTEYT, INPNNGGT, and ARYDFDVLFDV in sequence.

2. The antibody or antigen-binding fragment according to claim 1, characterized in that The light chain variable region of the antibody or antigen-binding fragment mAb21 comprises the amino acid sequence shown in SEQ ID NO:13, and the heavy chain variable region of the antibody or antigen-binding fragment mAb21 comprises the amino acid sequence shown in SEQ ID NO:

14.

3. The antibody or antigen-binding fragment according to claim 1, characterized in that The light chain variable region of the antibody or antigen-binding fragment mAb22 comprises the amino acid sequence shown in SEQ ID NO:15, and the heavy chain variable region of the antibody or antigen-binding fragment mAb22 comprises the amino acid sequence shown in SEQ ID NO:

16.

4. A hybridoma cell line mAb21 secreting anti-human PSP protein monoclonal antibody, characterized in that: The hybridoma cell line mAb21 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on November 20, 2024, with the deposit number CGMCC No.46136.

5. A hybridoma cell line mAb22 secreting anti-human PSP protein monoclonal antibody, characterized in that: The hybridoma cell line mAb22 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on November 20, 2024, with the deposit number CGMCC No.46137.

6. A monoclonal antibody against human PSP protein, characterized in that: The anti-human PSP protein monoclonal antibody is obtained by secreting the hybridoma cell line mAb21 according to claim 4 and / or is obtained by secreting the hybridoma cell line mAb22 according to claim 5.

7. A kit for detecting human PSP protein, characterized in that: The kit comprises a coating antibody and a detection antibody, wherein the coating antibody is the antibody mAb21 described in claim 1, and the detection antibody is the antibody mAb22 described in claim 1.

8. The kit according to claim 7, characterized in that The kit is a double antibody sandwich ELISA detection kit or an immunoassay kit prepared by chemiluminescence method.

9. Use of the antibody or antigen-binding fragment of claim 1 or the monoclonal antibody of claim 6 in detecting human PSP protein or preparing a kit for detecting human PSP protein.

10. Use of the hybridoma cell line according to claim 4 or 5 in preparing a monoclonal antibody for detecting human PSP protein.

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