Monoclonal Antibody Against Human PSP Protein and Its Application

Abstract

This application belongs to the field of immunoassay technology and discloses monoclonal antibodies against human PSP protein and their applications. The monoclonal antibodies against human PSP protein described herein can bind to human PSP protein with high specificity and affinity. The monoclonal antibodies against human PSP protein described herein can also be prepared into various immunoassay kits for detecting human PSP protein, particularly in immunoassay kits prepared using double-antibody sandwich ELISA or chemiluminescence methods, wherein the coating antibody is mAb21 and the detection antibody is mAb22. The detection sensitivity is less than 15 pg / ml, demonstrating 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 the body.

[0003] Depending on the source and preparation method, the molecular weight of PSP protein generally ranges from 25 to 30 kDa. PSP protein is primarily produced in the pancreas and intestines, but can also be expressed in other tissues such as the liver. In the human body, PSP protein is widely distributed in various tissues and organs, playing an important physiological role. PSP has a catalytic effect. PSP protein possesses phosphoserine phosphatase activity, catalyzing the dephosphorylation of O-phospho-L-serine to L-serine, the final step in L-serine biosynthesis. L-serine is a precursor for protein synthesis, the production of other amino acids, nucleotide metabolism, and glutathione synthesis. Therefore, PSP protein plays an important role in maintaining cellular 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 elevated, 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 that can secrete anti-human PSP monoclonal antibodies. It 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:

[0008] 1) an antibody or antigen-binding fragment mAb21, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, wherein the amino acid sequence of LCDR1-3 comprises SSVSY (SEQ ID NO: 1), DTS (SEQ ID NO: 2), and QQWSSNPPT (SEQ ID NO: 3), in sequence; and the heavy chain variable region comprises HCDR1-3, wherein the amino acid sequence of HCDR1-3 comprises GYTFSTYW (SEQ ID NO: 4), IFPGSGST (SEQ ID NO: 5), and VRKRYPYYFDY (SEQ ID NO: 6); and / or,

[0009] 2) an antibody or antigen-binding fragment mAb22, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, and the amino acid sequence of the LCDR1-3 comprises QSMSND (SEQ ID NO: 7), YAS (SEQ ID NO: 8), and QQDYSSPLT (SEQ ID NO: 9), in sequence; and the heavy chain variable region comprises HCDR1-3, and the amino acid sequence of the HCDR1-3 comprises GYTFTEYT (SEQ ID NO: 10), INPNNGGT (SEQ ID NO: 11), and ARYDFDVLFDV (SEQ ID NO: 12).

[0010] 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.

[0011] 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.

[0012] 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 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, preservation number: CGMCC No.46136.

[0013] 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 Culture Collection Administration of Microorganisms, 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, preservation number: CGMCC No.46137.

[0014] 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 secretion of the hybridoma cell line mAb22.

[0015] On the other hand, the present application also provides a kit for detecting human PSP protein, which comprises a coating antibody and a detection antibody, wherein the coating antibody is antibody mAb21, and the detection antibody is antibody mAb22.

[0016] Furthermore, the kit is a double antibody sandwich ELISA detection kit or an immunoassay kit prepared by chemiluminescence method.

[0017] 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.

[0018] 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.

[0019] Compared to existing technologies, the anti-human PSP monoclonal antibody of the present application can bind to human PSP protein with high specificity and affinity. The anti-human PSP monoclonal antibody of the present application can also be prepared into various immunoassay kits for detecting human PSP protein, particularly in immunoassay kits prepared using double-antibody sandwich ELISA or chemiluminescence, wherein the coating antibody is antibody mAb21 and the detection antibody is antibody mAb22. The detection sensitivity is less than 15 pg / ml, which has high clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The double antibody sandwich ELISA method is used to detect the human PSP standard antigen. The horizontal axis is the concentration of the human PSP standard antigen (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

[0021] The present invention will be further described below with reference to specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. Experimental procedures in the following examples, where specific conditions are not specified, were generally performed under conventional conditions, conditions described in laboratory manuals, or conditions recommended by the manufacturer.

[0022] Example 1 Preparation of anti-human PSP monoclonal antibodies

[0023] 1) Preparation of human PSP immunogen

[0024] The PSP gene NM_002909, encoding the full 166 amino acids (aa) of PSP, was selected from Genebank. The immunogen of the antibody described herein is PSP 1-160aa. This gene was cloned into the pET23a expression plasmid with a histag at the C-terminus and expressed in BL21 Escherichia coli. Purification was performed using a nickel column and the purity was assessed by SDS-PAGE electrophoresis. After electrophoresis, a target protein band with a molecular weight of approximately 18 kDa was observed on the gel, indicating a purity exceeding 90%, meeting the purity requirements for monoclonal antibody preparation.

[0025] 2) Animal immunization

[0026] The purified PSP recombinant protein was emulsified in complete Freund's adjuvant and administered subcutaneously or intraperitoneally to 6-8-week-old BALB / c mice at a dose of 80 μg per mouse. Two weeks later, a second immunization was performed using incomplete Freund's adjuvant at a dose of 60 μg per mouse. After both immunizations, tail blood was collected and serum titers were determined by serial dilution ELISA. Based on the titer results, a booster immunization was determined, and mice with the highest antibody titer were selected for cell fusion.

[0027] 3) Cell fusion and clone screening

[0028] 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 fused with 50% PEG 4000. After centrifugation, resuspend them in HAT selective medium and inoculate them into 96-well microplates containing feeder cells. Culture them in a 37°C, 5% CO2 incubator. Replace half of the HAT medium on the 4th and 7th days after fusion, and replace it with HT medium after 10 days. Observe the growth of hybridoma cells. When the clones grow to 1 / 4 to 1 / 3 of the bottom area of the well, take the culture supernatant, perform antibody detection by ELISA, and screen the positive clone wells. Use the limiting dilution method to clone and culture 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 (i.e., the ELISA titer is 1:10). 4 The above positive clones) were used to obtain hybridoma cell lines mAb21, mAb22 and mAb23 that secreted anti-human PSP protein monoclonal antibodies.

[0029] 4) Preparation and purification of monoclonal antibodies

[0030] Hybridoma cell lines mAb21 and mAb22 were cultured in suspension in serum-free medium. After approximately one week, when the cell supernatant volume reached the desired volume and the cell mortality rate reached 60%-70%, the cell suspension was harvested and centrifuged to obtain the supernatant. Antibodies were purified by affinity chromatography using a column material selected based on the antibody subtype. The purified monoclonal antibody concentration was determined by the BCA assay, and the purified antibody was aliquoted and frozen to obtain monoclonal antibodies mAb21, mAb22, and mAb23.

[0031] Example 2 Identification of anti-human PSP monoclonal antibodies

[0032] 1) Determination of affinity constant of monoclonal antibodies

[0033] The affinity constant (Ka) was determined by non-competitive ELISA.

[0034] 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.

[0035] Blocking: Wash the plate 4 times with PBST, add BSA solution at 200 μL / well, and incubate at 37°C for 2 h;

[0036] Adding monoclonal antibodies: Wash the plate four times with PBST, dilute the monoclonal antibody OTIR3A11 starting at 100 μg / mL in carbonate buffer, add 100 μL to each well, and incubate at 37°C for 2 h;

[0037] Add enzyme-labeled secondary antibody: wash the plate four times with PBST, add 100 μL of 1:10000 diluted HRP enzyme-labeled Ig secondary antibody to each well, and incubate at 37°C for 30 min;

[0038] Color development and termination: Wash the plate four 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;

[0039] Detection: Measure the absorbance value at a wavelength of 450 nm (A450nm).

[0040] The logarithm of the antibody concentration was used as the horizontal axis and the OD value was used as the vertical axis to draw an S-shaped curve. After calculation, the affinity constants Ka of the 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.

[0041] 2) Antibody pairing

[0042] To select the optimal combination of coating and detection antibodies, a checkerboard pattern was used to coat ELISA plates with the three purified monoclonal antibodies mAb21, mAb22, and mAb23 overnight at 4°C. The next day, the plates were removed, washed once with PBST, blocked with 1% BSA solution at 37°C for 2 hours, and washed three times with PBST. 100 μl of full-length human PSP protein at a concentration of 1 ng / ml was added to each well and incubated at 37°C for 1 hour. After incubation, the plates were removed and washed three times with PBST. HRP-conjugated versions of the three antibodies were added as detection antibodies. Double-antibody sandwich ELISAs were performed, pairing the three monoclonal antibodies with their corresponding enzyme-conjugated monoclonal antibodies and incubating at 37°C for 1 hour. After washing five times with PBST, TMB substrate was added and color was developed at 37°C for 10 minutes. Stop solution was added and OD450 readings were measured on a microplate reader. The optimal monoclonal antibody pair was selected based on the sample OD value and the background value of the negative control. The results of the pairing screening are shown in Table 1.

[0043] Table 1 Antibody pairing experiment screening results

[0044]

[0045] It can be seen that the antibody mAb21 involved in this application is the best as the coating antibody, and mAb22 is the best as the detection antibody.

[0046] Example 3 Analysis of the Gene and Amino Acid Sequences of the Variable Regions of Monoclonal Antibodies mAb21 and mAb22. Recombinant plasmids of mAb21 and mAb22 antibodies were used as DNA templates. Sequencing primers for the light and heavy chain variable regions were designed based on the vector sequences at the 5' ends of the light and heavy chains of the templates. Sequencing was performed using an ABI 3730 sequencer. The nucleotide sequences of the light and heavy chain variable regions of the monoclonal antibodies mAb21 and mAb22 were obtained through sequencing.

[0047] The nucleotide sequences of the light chain variable region and the heavy chain variable region were sequenced and analyzed using IMGT / V-QUEST analysis software at http: / / www.imgt.org on the Internet. The monoclonal antibody mAb21 comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, which in turn comprise the amino acid sequences shown in SEQ ID NOs: 1-3, respectively; and the heavy chain variable region comprises HCDR1-3, which in turn comprise the amino acid sequences shown in SEQ ID NOs: 4-6, respectively.

[0048] The light chain variable region of monoclonal antibody mAb21 comprises the amino acid sequence shown in SEQ ID NO:13. The total length of VL is 107 amino acids. The amino acid numbers of the four FR domains are 26, 17, 31, and 11, respectively. The amino acid numbers of the three LCDR domains are 5, 3, and 8, respectively. The regions of LCDR1, LCDR2, and LCDR3 are 27aa-31aa, 49aa-51aa, and 88aa-96aa, respectively. Their amino acid sequences are: SSVSY (SEQ ID NO.1), DTS (SEQ ID NO.2), and QQWSSNPPT (SEQ ID NO.3), respectively.

[0049] The heavy chain variable region of 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 FR domains are 24, 17, 36, and 11, respectively. The amino acid numbers of the three HCDR domains are 8, 8, and 11, respectively. The HCDR1, HCDR2, and HCDR3 are 25aa-32aa, 50aa-57aa, and 94aa-104aa, respectively. Their amino acid sequences are GYTFSTYW (SEQ ID NO.4), IFPGSGST (SEQ ID NO.5), and VRKRYPYYFDY (SEQ ID NO.6), respectively.

[0050] The monoclonal antibody mAb22 includes a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises LCDR1-3, and the LCDR1-3 respectively comprise the amino acid sequences shown in SEQ ID NOs: 7-9, and the heavy chain variable region comprises HCDR1-3, and the HCDR1-3 respectively comprise the amino acid sequences shown in SEQ ID NOs: 10-12.

[0051] The light chain variable region of monoclonal antibody mAb22 comprises the amino acid sequence set forth in SEQ ID NO:15. The VL is 107 amino acids in length. The four FR domains contain 26, 16, 36, and 11 amino acids, respectively. The three LCDR domains contain 6, 3, and 9 amino acids, respectively. The LCDR1, LCDR2, and LCDR3 regions 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), and QQDYSSPLT (SEQ ID NO.9).

[0052] 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 FR domains are 24, 17, 36, and 11, respectively. The amino acid numbers of the three HCDR domains are 8, 8, and 11, respectively. The HCDR1, HCDR2, and HCDR3 are 25aa-32aa, 50aa-57aa, and 94aa-104aa, respectively. Their amino acid sequences are GYTFTEYT (SEQ ID NO. 10), INPNNGGT (SEQ ID NO. 11), and ARYDFDVLFDV (SEQ ID NO. 12), respectively.

[0053] Example 4: Anti-human PSP monoclonal antibodies are used to prepare a double antibody sandwich ELISA detection kit. A human PSP detection kit is prepared based on the ELISA double antibody sandwich method for clinical auxiliary diagnosis of diseases related to human PSP.

[0054] 1) Kit Composition

[0055] 1. Coating the strips with mAb21: Dilute the antibody to 1 μg / ml in PBS buffer and coat 100 μl per well on a microplate. Incubate overnight at 4°C, wash three times with PBST, and spin dry. Block with PBS containing 1% BSA, 5% sucrose, and 0.05% Proclin 300 at 37°C for 2 h. Discard the blocking solution and spin dry. Place the coated plate in a 37°C oven for 2 h to complete the coating. Seal the plate in aluminum foil and store at 4°C until needed.

[0056] 2. Reagent Preparation

[0057] 2.1 Preparation of enzyme conjugates The anti-human PSP detection antibody mAb22 was labeled using a simple sodium periodate method. 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, and the solution was sterilized by filtration.

[0058] 2.2 The washing buffer was conventional PBST at pH 7.4, containing 0.05% Proclin 300, and prepared into a 20-fold concentrated solution.

[0059] 2.3 Enzyme substrate solution (developing solution): single-component TMB (purchased from the market).

[0060] 2.4 Sample dilution solution: PBS containing 1% BSA and 0.05% Proclin 300, filter sterilize.

[0061] 2.5 Stop solution: Add 10 ml of HCl (36-38%) to 110 ml of distilled water and mix slowly to make 1N HCl.

[0062] 2.6 Standard: Full-length human PSP protein. Dilute the sample to 20 μg / ml using PBS containing 1% BSA, 5% sucrose, 10% glycerol, and 0.05% Proclin 300. Filter sterilize and aliquot aseptically.

[0063] 2) Key points of detection operation

[0064] 1. To ensure the accuracy of the test results, it is recommended to set up double-well measurement for both standards and samples. A standard curve should be prepared for each test.

[0065] 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 when calculating.

[0066] 3. Sample addition: When adding samples, please use clean, disposable pipette tips to avoid cross-contamination. Add samples gently to avoid foaming. Add samples to the bottom of the wells of the ELISA plate, not along the well walls.

[0067] 4. Incubation: To prevent sample evaporation or contamination, the ELISA plate must be covered with a plate sticker during incubation. The plate should not be allowed to dry out during the experiment. During incubation, the incubator temperature should be monitored to ensure it remains constant at 37°C and adjusted promptly. The incubator should not be opened too many times during incubation to avoid disrupting temperature balance.

[0068] 5. Washing: The washing process is very important. Insufficient washing can easily cause false positives.

[0069] 6. Color Development: To ensure the accuracy of the experimental results, the stop solution should be added as soon as the substrate reaction time expires. After adding the substrate solution, the color development can be observed at regular intervals (e.g., every 10 minutes) to control the reaction time. When a clear blue gradient is visible in the first 3-4 wells of the standard sample and the color development is less obvious in the last 3-4 wells, the stop solution can be added to terminate the reaction. The blue color will immediately turn yellow. The stop solution should be added in the same order as the substrate solution.

[0070] 7. The substrate solution should be light blue or colorless. If it becomes significantly darker, it must be discarded. The substrate solution is easily contaminated; please store it away from light.

[0071] 3) Determination of standard curves for different dilutions of standard human PSP protein

[0072] The prepared double-antibody sandwich ELISA kit for detecting human PSP protein was removed from a 4°C refrigerator and equilibrated to room temperature. The standard was diluted with PBS from 15 ng / ml to 250 pg / ml to prepare six concentration series of standards: 0, 15, 30, 60, 125, and 250. According to the above detection method, 100 μl of the standard was added to each well and incubated. The liquid was then discarded, and HRP-labeled detection antibody working solution was added and incubated. The liquid in the wells was discarded, and the substrate solution was added to each well after drying. After color development in the dark, stop solution was added to each well. Within 5 minutes after the reaction was terminated, the optical density (OD) of each well was measured sequentially at a wavelength of 450 nm using a microplate reader to create a standard curve. The results are shown in Table 2.

[0073] Table 2 Detection of human PSP standards by double antibody sandwich ELISA

[0074]

[0075]

[0076] According to Table 2, the PSP detection standard curve was prepared. Figure 1 The R of the standard curve is shown. 2 =0.9956, linear detection range 0-1000pg / mL, sample concentration calculation formula is derived: y=0.0073x+0.1217, detection sensitivity is less than 15pg / ml.

[0077] 4) Sample testing

[0078] The above method was used to test sera from sepsis patients, and the results are shown in Table 3. Based on the results, the mAb21 antibody of the present invention, used as the coating antibody, and the mAb22 antibody as the detection antibody, were detected by magnetic microparticles and showed good correlation with clinical comparisons. This kit provides an auxiliary means for the diagnosis of diseases related to the PSP protein.

[0079] Table 3 Sample testing

[0080]

[0081] 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 non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

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, comprising 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 LCDR1-3 are, in order: SSVSY, DTS, QQWSSNPPT; and the heavy chain variable region comprises HCDR1-3, wherein the amino acid sequences of HCDR1-3 are, in order: GYTFSTYW, IFPGSGST, VRKRYPYYFDY; and / or, 2) 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 LCDR1-3 are QSMSND, YAS, and QQDYSSPLT, respectively; and the heavy chain variable region comprises HCDR1-3, and the amino acid sequences of HCDR1-3 are GYTFTEYT, INPNNGGT, and ARYDFDVLFDV, respectively.

2. The antibody or antigen-binding fragment according to claim 1, wherein 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, wherein 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 the China 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 the China 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 secretion of the hybridoma cell line mAb21 according to claim 4 and / or is obtained by secretion of 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 according to claim 1, and the detection antibody is the antibody mAb22 according to 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 according to claim 1 or the monoclonal antibody according to claim 6 in the preparation of a kit for detecting human PSP protein.

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

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