Anti-human cd61 antibodies and uses thereof

Abstract

The application discloses an anti-human CD61 antibody and application thereof, and the antibody comprises a heavy chain variable region and a light chain variable region; CDR1, CDR2 and CDR3 of the light chain variable region are respectively amino acid sequences as shown in SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3; and CDR1, CDR2 and CDR3 of the heavy chain variable region are respectively amino acid sequences as shown in SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6. The antibody of the application has the same binding activity as a commercially available antibody under the same concentration, and the good antigen binding capacity is verified; the antibody of the application has higher fluorescence intensity and stronger affinity, and is suitable for trace sample detection or low-dose diagnostic reagents.

Description

Technical Field

[0001] This invention belongs to the field of in vitro diagnostic technology, and in particular relates to an anti-human CD61 antibody and its application. Background Technology

[0002] CD61 (platelet glycoprotein GPIIIa) is the β3 subunit of integrin, with a molecular weight of 105-110 kDa, and is mainly expressed in platelets, megakaryocytes, osteoclasts, and vascular endothelial cells. CD61 binds to the integrin αIIb chain (CD41) to form the GPIIb / IIIa complex (αIIb / β3 integrin), participating in the function of high-affinity receptors for platelets and fibrinogen, playing a crucial role in coagulation and thrombosis. As a platelet-specific antigen, CD61 is commonly used in the diagnosis and monitoring of diseases such as thrombocytopenia and myelodysplastic syndromes. It is also involved in bone marrow biopsy analysis for diseases such as megakaryocytic leukemia, serving as a reference indicator for the degree of decalcification.

[0003] Pancreatic cancer is a malignant tumor originating from the epithelial cells and acinar cells of the pancreatic ducts, often referred to as the "king of cancers" in the medical community. Its causes are not fully understood, but research suggests that long-term smoking, poor dietary habits, high body fat percentage, and chronic pancreatic damage may increase the risk. Symptoms of pancreatic cancer typically appear in later stages, including jaundice, weight loss, and abdominal discomfort or pain. Data shows that over 80% of patients are diagnosed at a locally advanced or metastatic stage, missing the optimal time for surgery. Furthermore, pancreatic cancer has an extremely high mortality rate, making it a leading cause of cancer death. Traditional surgical resection is the only potentially curative treatment, but it is only suitable for a small number of early-stage patients. Most patients are diagnosed at a point when surgery is no longer an option, relying instead on palliative treatments such as chemotherapy and radiotherapy, with often unsatisfactory outcomes.

[0004] There are two distinct types of cancer cells in pancreatic cancer: one is the docile epithelial cells responsible for tumor growth, and the other is the rebellious stromal cells that dominate metastasis and drug resistance. The latter is a key factor leading to the extremely poor prognosis of pancreatic cancer, and these two types of cells can also transform into each other. Studies have found that epithelial cancer cells secrete a key protein called SPP1 to maintain the survival of stromal cells. Clinical data also confirms that significantly elevated SPP1 levels in the plasma of advanced patients are closely related to disease progression. In mouse model experiments, once the SPP1 gene is knocked out, stromal cancer cells almost completely disappear, and the tumor not only shrinks but also stops metastasizing. Conversely, if SPP1 is overexpressed, a large number of epithelial cells will transform into stromal cells, resulting in a surge in the tumor's metastatic ability. Summary of the Invention

[0005] In view of this, the present invention aims to provide an anti-human CD61 antibody and its application to solve at least one technical problem in the background art.

[0006] To achieve the above objectives, the technical solution of the present invention is implemented as follows: An anti-human CD61 antibody, comprising a heavy chain variable region and a light chain variable region; The CDR1, CDR2, and CDR3 of the light chain variable region are the amino acid sequences shown in SEQ ID NO.1, SEQ ID NO.2, and SEQ ID NO.3, respectively. The amino acid sequence of SEQ ID NO.1 is: TASSSVSSSNLH; The amino acid sequence of SEQ ID NO.2 is: RTSNLAS; The amino acid sequence of SEQ ID NO.3 is: HQYHRSPLT.

[0007] The CDR1, CDR2, and CDR3 of the heavy chain variable region are the amino acid sequences shown in SEQ ID NO.4, SEQ ID NO.5, and SEQ ID NO.6, respectively.

[0008] The amino acid sequence of SEQ ID NO.4 is: GYTFTSYWIH; The amino acid sequence of SEQ ID NO.5 is: YINPSTGHSEYNQKFKD; The amino acid sequence of SEQ ID NO.6 is: WTTEGA.

[0009] Furthermore, the FR1, FR2, FR3, and FR4 of the light chain variable region are the amino acid sequences shown in SEQ ID NO.7, SEQ ID NO.8, SEQ ID NO.9, and SEQ ID NO.10, respectively; The amino acid sequence of SEQ ID NO.7 is: QIVLTQSPEIMSASLGERVTMTC; The amino acid sequence of SEQ ID NO.8 is: WFQQKPGSSPKLWIY; The amino acid sequence of SEQ ID NO.9 is as follows: GVPPRFSGSGSGTSYSLTISSMEAEDAATYYC; The amino acid sequence of SEQ ID NO.10 is: FGAGTKLELKR.

[0010] The FR1, FR2, FR3, and FR4 of the heavy chain variable region are the amino acid sequences shown in SEQ ID NO.11, SEQ ID NO.12, SEQ ID NO.13, and SEQ ID NO.14, respectively.

[0011] The amino acid sequence of SEQ ID NO.11 is as follows: QVQLQQSGAELAKPGASVKMSCKAS; The amino acid sequence of SEQ ID NO.12 is: WVKQRPGQGLEWIG; The amino acid sequence of SEQ ID NO.13 is as follows: KATLTADKSSSTAYMQLNSLTSEDSAVYYCGR; The amino acid sequence of SEQ ID NO.14 is: WGQGTTLTVSS.

[0012] Furthermore, the nucleotide sequence of the light chain variable region of the antibody is shown in SEQ ID NO.15, and the nucleotide sequence of the heavy chain variable region is shown in SEQ ID NO.16.

[0013] The nucleotide sequence of SEQ ID NO.15 is as follows: CAAATTGTTCTCACCCAGTCTCCAGAGATCATGTCTGCATCTCTAGGGGAACGGGTCACCATGACCTGCACTGCCAGCTCAAGTGTAAGTTCCAGTAACTTGCACTGGTTCCAGCAGAAGCCAGGATCCCTCCCCCAAACTCTGGATTTATAGAACATCCAACCT GGCCTCTGGAGTCCCACCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCACCAGTATCATCGTTCCCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGG; The nucleotide sequence of SEQ ID NO.16 is as follows: CAGGTCCAGCTTCAGCAATCTGGGGCTGAACTGGCAAAACCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACTAGCTACTGGATCCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGATACATTAATCCTAGCACTGGTCATAG TGAATATAATCAGAAATTCAAGGACAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGCCTATATGCAACTGAACAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGGAAGATGGACTACGGAGGGCGCCTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA.

[0014] The amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO.17, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO.18.

[0015] The amino acid sequence of SEQ ID NO.17 is as follows: QIVLTQSPEIMSASLGERVTMTCTASSSVSSSNLHWFQQKPGSSPKLWIYRTSNLASGVPPRFSGSGSGTSYSLTISSMEAEDAATYYCHQYHRSPLTFGAGTKLELKR; The amino acid sequence of SEQ ID NO.18 is as follows: QVQLQQSGAELAKPGASVKMSCKASGYTFTSYWIHWVKQRPGQGLEWIGYINPSTGHSEYNQKFKDKATLTADKSSSTAYMQLNSLTSEDSAVYYCGRWTTEGAWGQGTTLTVSS.

[0016] A nucleic acid molecule that encodes the aforementioned anti-human CD61 antibody.

[0017] The nucleic acid molecule comprises a nucleotide sequence encoding the heavy chain variable region of the anti-human CD61 antibody and a nucleotide sequence encoding the light chain variable region.

[0018] An expression vector comprising the aforementioned nucleic acid molecule.

[0019] A cell containing one of the aforementioned nucleic acid molecules or the aforementioned expression vectors.

[0020] An immunoassay kit comprising the aforementioned anti-human CD61 antibody.

[0021] The above-mentioned application of the anti-human CD61 antibody, specifically its application in the preparation of drugs for treating pancreatic cancer.

[0022] Compared with existing technologies, the anti-human CD61 antibody and its application described in this invention have the following advantages: 1. The CDR and FR sequences of the antibody light and heavy chain variable regions in this application have been identified, which facilitates subsequent engineering modification and production.

[0023] 2. This application has high purity and complete structure, and the electrophoresis image ( Figure 1 The results showed clear bands with no obvious impurities under both reducing and non-reducing conditions, indicating a mature purification process and stable antibody structure. The activity validation was thorough, showing that the antibody in this application has comparable binding activity to commercially available antibodies at the same concentration, validating its excellent antigen-binding ability. Furthermore, the fluorescence intensity of this application is higher, making it suitable for micro-sample detection or low-dose diagnostic reagents.

[0024] 3. The antibody in this application has high specificity and can be used to develop CD61-targeted therapeutic drugs, especially suitable for tumors such as pancreatic cancer with abnormal activation of the SPP1 / CD61 signaling pathway. Attached Figure Description

[0025] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 The images shown are electrophoresis diagrams of the purified anti-human CD61 antibody in Example 1 of this invention (a is a non-reduction electrophoresis diagram, b is a reduction electrophoresis diagram). Figure 2 This is a detection graph of the activity of the anti-human CD61 antibody in Example 5 of the present invention; Figure 3 This is a graph showing the activity of CD61-PE fluorescent antibody after anti-human CD61-labeled fluorescein PE. Detailed Implementation

[0026] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0027] The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0028] Example 1: Coating of CD61 plasmid with lentivirus and preparation of immune cells; 1) Lentiviral coating: A three-plasmid packaging system was used, consisting of one packaging plasmid (psPAX2), one envelope plasmid (pMD2.G), and one expression plasmid. The nucleotide sequence of human CD61 antigen (GenBank: BC127666.1) was synthesized into the target plasmid pCDH-CMV-MCS-EF1-Puro. The psPAX2 packaging plasmid, pMD2.G envelope plasmid, and pCDH-CMV-MCS-EF1-Puro expression plasmid containing the target gene were extracted separately and mixed at a ratio of psPAX2 packaging plasmid: pMD2.G envelope plasmid: pCDH-CMV-MCS-EF1-Puro expression plasmid containing the target gene = 3:1:4. The plasmid was prepared at 1 μg / mL and mixed with PEI at a mass ratio of 3:1. After standing for 10 minutes, it was added to 293 cells. Forty-eight hours later, the viral supernatant was collected, and 10 mL of pre-warmed 293 medium was added to the cell culture dish. The collected viral supernatant was placed in a 4°C freezer. The 72-hour viral supernatant was collected and mixed with the 48-hour viral supernatant. The mixture was centrifuged at 40,000 rpm for 120 min to concentrate the virus, aliquoted, and stored at -80°C.

[0029] 2) Lentiviral infection of 3T3 cells: Observe the status of 3T3 cells. If the cell viability is higher than 90%, seed them into 24-well culture plates. After culturing for 24 hours, add Polybrene at a working concentration of 5-10 μg / mL, then add 200 μL of lentivirus, mix well, and place in a 37℃ incubator. Observe the fluorescence rate of 3T3 cells every day. When the fluorescence rate is greater than 90%, expand the cells for culture and use them as immunogens for mouse immunization.

[0030] Example 2: Animal immunization; 1) Expand the culture of CD61-3T3 transfected cells by intraperitoneal injection at a dose of 1×10⁻⁶. 6 Cells per mouse were used to immunize 4-week-old female BALB / c mice; immunization was repeated every 2 weeks for a total of 3 immunizations. Tail blood was collected from the mice, and ELISA was used to detect the titer. The mouse with the highest titer was selected for subsequent cell fusion.

[0031] Example 3: Cell fusion and screening; 1) Cell preparation: Select mice with the highest titer and administer a shock immunization via intraperitoneal injection of transfected cell lines at a dosage of 5 × 10⁶ cells / mice. After 3 days, perform fusion, remove the spleen from the mice under sterile conditions, grind the spleen, collect spleen cells through a sieve, and wash once with PBS. At the same time, collect myeloma cells in the logarithmic growth phase and mix myeloma cells with spleen cells at a ratio of 1:5. Centrifuge at 1000 rpm for 5 min and discard the supernatant.

[0032] 2) Induction of fusion: Add 1 mL of PEG solution as an inducer, shake gently, and incubate at 37°C for 1 min to allow mouse spleen cells and mouse myeloma cells to fuse.

[0033] 3) Screening hybrid cells: Inoculate the hybrid cells into a selective medium (such as HAT medium), discard unfused cells and homonuclear fusion cells, and retain heteronuclear hybrid cells.

[0034] 4) Clonal culture and identification: Positive clone wells were screened using flow cytometry, and three rounds of subcloning were performed using the limiting dilution method. The activity of the cell supernatant was identified using flow cytometry, and the clones with the best activity were selected for expansion culture for later use.

[0035] Example 4: Preparation of anti-human CD61 antibody; 1) Preparation of CD61 mouse ascites: Hybridoma cells in good growth condition were collected and injected intraperitoneally into mice at a concentration of 1×10⁻⁶. 6 Cells per mouse branch; when the mouse abdomen is noticeably distended and the skin is taut and shiny, ascites can be collected in batches; the collected ascites is stored at -20℃.

[0036] 2) Purification of anti-human CD61 antibody: The harvested ascites fluid was centrifuged at 10,000 rpm for 30 min, filtered through a 0.45 μm filter membrane, and then purified using a Protein A column. The column was first equilibrated with PBS, then loaded with the sample, and finally eluted with glycine at pH 3.5. The solution was then replaced with G25 molecular sieve buffer, and finally replaced with PBS buffer. 100 μL of the solution was taken for protein electrophoresis identification. Figure 1 As shown, a is the non-reduction electrophoresis pattern of the anti-human CD61 antibody, and b is the reduction electrophoresis pattern of the anti-human CD61 antibody. The antibody in this application has high purity and a complete structure. (The electrophoresis patterns are...) Figure 1 The results showed that the antibody bands were clear under both reducing and non-reducing conditions, with no obvious impurities, indicating that the purification process was mature and the antibody structure was stable.

[0037] The nucleotide sequence of the light chain variable region of the above-mentioned anti-human CD61 antibody is shown in SEQ ID NO.15: SEQ ID NO.15: CAAATTGTTCTCACCCAGTCTCCAGAGATCATGTCTGCATCTCTAGGGGAACGGGTCACCATGACCTGCACTGCCAGCTCAAGTGTAAGTTCCAGTAACTTGCACTGGTTCCAGCAGAAGCCAGGATCCCTCCCCCAAACTCTGGATTTATAGAACATCCAACCT GGCCTCTGGAGTCCCACCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCACCAGTATCATCGTTCCCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGG; The nucleotide sequence of the heavy chain variable region of the CD61 antibody is shown in SEQ ID NO.16: CAGGTCCAGCTTCAGCAATCTGGGGCTGAACTGGCAAAACCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTACTAGCTACTGGATCCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGATACATTAATCCTAGCACTGGTCATAG TGAATATAATCAGAAATTCAAGGACAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGCCTATATGCAACTGAACAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGGAAGATGGACTACGGAGGGCGCCTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA; The amino acid sequence of the light chain variable region of the CD61 antibody is shown in SEQ ID NO.17, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO.18.

[0038] The amino acid sequence of SEQ ID NO.17 is as follows: QIVLTQSPEIMSASLGERVTMTCTASSSVSSSNLHWFQQKPGSSPKLWIYRTSNLASGVPPRFSGSGSGTSYSLTISSMEAEDAATYYCHQYHRSPLTFGAGTKLELKR; The amino acid sequence of SEQ ID NO.18 is as follows: QVQLQQSGAELAKPGASVKMSCKASGYTFTSYWIHWVKQRPGQGLEWIGYINPSTGHSEYNQKFKDKATLTADKSSSTAYMQLNSLTSEDSAVYYCGRWTTEGAWGQGTTLTVSS.

[0039] CD61 Light Chain: The amino acid sequence of FR1 in the light chain variable region is SEQ ID NO.7: The amino acid sequence of SEQ ID NO.7 is: QIVLTQSPEIMSASLGERVTMTC; The CDR1 of the light chain variable region has the amino acid sequence SEQ ID NO.1; The amino acid sequence of SEQ ID NO.1 is: TASSSVSSSNLH; The amino acid sequence of FR2 in the light chain variable region is SEQ ID NO. 8; The amino acid sequence of SEQ ID NO.8 is: WFQQKPGSSPKLWIY; The CDR2 of the light chain variable region has the amino acid sequence SEQ ID NO.2; The amino acid sequence of SEQ ID NO.2 is: RTSNLAS; FR3, a variable region of the light chain, has the amino acid sequence SEQ ID NO.9; The amino acid sequence of SEQ ID NO.9 is as follows: GVPPRFSGSGSGTSYSLTISSMEAEDAATYYC; CDR3: HQYHRSPLT; The CDR3 of the light chain variable region has the amino acid sequence SEQ ID NO.3; The amino acid sequence of SEQ ID NO.3 is: HQYHRSPLT.

[0040] FR4, a variable region of the light chain, has the amino acid sequence SEQ ID NO.10; The amino acid sequence of SEQ ID NO.10 is: FGAGTKLELKR.

[0041] CD61 heavy chain; The heavy chain variable region FR1, the amino acid sequence of FR1 is SEQ ID NO.11; The amino acid sequence of SEQ ID NO.11 is as follows: QVQLQQSGAELAKPGASVKMSCKAS; The heavy chain variable region CDR1, the amino acid sequence of CDR1 is SEQ ID NO.4; The amino acid sequence of SEQ ID NO.4 is: GYTFTSYWIH; The heavy chain variable region FR2 has the amino acid sequence SEQ ID NO.12; The amino acid sequence of SEQ ID NO.12 is: WVKQRPGQGLEWIG; The heavy chain variable region CDR2, the amino acid sequence of CDR2 is SEQ ID NO.5; The amino acid sequence of SEQ ID NO.5 is: YINPSTGHSEYNQKFKD; FR3 of the heavy chain variable region, the amino acid sequence of FR3 is SEQ ID NO.13; The amino acid sequence of SEQ ID NO.13 is as follows: KATLTADKSSSTAYMQLNSLTSEDSAVYYCGR; The heavy chain variable region CDR3 has the amino acid sequence SEQ ID NO.6; The amino acid sequence of SEQ ID NO.6 is: WTTEGA.

[0042] The heavy chain variable region FR4 has the amino acid sequence SEQ ID NO.14; The amino acid sequence of SEQ ID NO.14 is: WGQGTTLTVSS.

[0043] Example 5: Determination of the activity of anti-human CD61 antibody; 1) Platelet detection: Take 5 mL of EDTA-K2 anticoagulated peripheral blood, centrifuge at 600 rpm for 10 minutes, take 300 µl of the total volume of the upper plasma layer, and resuspend in PBS buffer to 1 mL. Add 1.0 μg, 0.5 μg, and 0.25 μg of the CD61 antibody prepared in Example 4 and the CD61 antibody (clone number F11 purchased from Thermo) to each tube. Add 100 µl of platelet suspension to each tube and incubate at room temperature in the dark for 20 minutes. Add 1 mL of PBS to each tube, vortex thoroughly, centrifuge at 1200 rpm for 5 minutes, and discard the supernatant. Dilute Tonbo secondary antibody (catalog number 35-8055-U500) 1:700, add 50 µl of the diluted secondary antibody to each tube, and vortex thoroughly. Incubate at room temperature in the dark for 20 minutes. Add 1 mL of PBS to each tube, vortex thoroughly, centrifuge at 1200 rpm for 5 minutes, and discard the supernatant. Add 250 µl of PBS to each tube, vortex thoroughly, and perform flow cytometry analysis. The activity detection results of the anti-human CD61 antibody are as follows: Figure 2 As shown; from Figure 2 It can be concluded that when the antibody dosage is 0.25 μg, the fluorescence intensity value of the CD61 antibody of this application is higher than that of the commercially available CD61 antibody; when the antibody dosage is 0.50 μg and 1.00 μg, the fluorescence intensity value of the commercially available CD61 antibody is higher than that of the CD61 antibody of this application.

[0044] Example 6: Anti-human CD61-PE labeling; 1) Take CD61 antibody, add TCEP to a final concentration of 5-10 mmol / L, react at 25℃ for 1 hour, then mix it with activated fluorescein PE (product number HA-201-500) purchased from Aoki, stir at 25℃, and react for 1-2 hours; then purify using an S200 molecular sieve purification column, after equilibration, loading, and elution to obtain CD61-PE labeled antibody.

[0045] Example 7: Determination of the activity of anti-human CD61-PE antibody; Platelet count: Take 5 mL of EDTA-K2 anticoagulated peripheral blood, centrifuge at 600 rpm for 10 minutes, and take 300 µL of the supernatant plasma. Resuspend in PBS buffer to 1 mL. Add 1.0 μg, 0.5 μg, and 0.25 μg of the antibody CD61-PE prepared in Example 4 and the purchased antibody CD61-PE, respectively. Add 100 µL of platelet suspension to each tube and react at room temperature in the dark for 20 minutes. Add 1 mL of PBS to each tube, vortex thoroughly, centrifuge at 1200 rpm for 5 minutes, and discard the supernatant. Add 250 µL of PBS to each tube, vortex thoroughly, and use flow cytometry to detect the activity of the CD61-PE fluorescent antibody after anti-human CD61-labeled fluorescein PE as follows. Figure 3 As shown. From Figure 3It can be concluded that: when the antibody dosage is 0.25 μg to 0.50 μg, the fluorescence intensity value of the CD61 antibody of this application is higher than that of commercially available CD61 antibodies; when the antibody dosage is 0.50 μg to 1.00 μg, the fluorescence intensity value of commercially available CD61 antibodies is higher than that of the CD61 antibody of this application. At low concentrations, the fluorescent labeling efficiency or affinity of this application is higher, making it suitable for micro-sample detection or low-dose diagnostic reagents.

[0046] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An anti-human CD61 antibody, characterized in that: Includes the variable regions of heavy chains and light chains; The CDR1, CDR2, and CDR3 of the light chain variable region are the amino acid sequences shown in SEQ ID NO.1, SEQ ID NO.2, and SEQ ID NO.3, respectively. The CDR1, CDR2, and CDR3 of the heavy chain variable region are the amino acid sequences shown in SEQ ID NO.4, SEQ ID NO.5, and SEQ ID NO.6, respectively.

2. The anti-human CD61 antibody according to claim 1, characterized in that: The FR1, FR2, FR3, and FR4 of the light chain variable region are the amino acid sequences shown in SEQ ID NO.7, SEQ ID NO.8, SEQ ID NO.9, and SEQ ID NO.10, respectively. The FR1, FR2, FR3, and FR4 of the heavy chain variable region are the amino acid sequences shown in SEQ ID NO.11, SEQ ID NO.12, SEQ ID NO.13, and SEQ ID NO.14, respectively.

3. The anti-human CD61 antibody according to claim 1, characterized in that: The nucleotide sequence of the light chain variable region is shown in SEQ ID NO.15, and the nucleotide sequence of the heavy chain variable region is shown in SEQ ID NO.

16.

4. A nucleic acid molecule, characterized in that: The nucleic acid molecule encodes the anti-human CD61 antibody as described in any one of claims 1-3.

5. The nucleic acid molecule according to claim 4, characterized in that: The nucleic acid molecule comprises a nucleotide sequence encoding the heavy chain variable region of the anti-human CD61 antibody and a nucleotide sequence encoding the light chain variable region.

6. An expression vector, characterized in that: The expression vector contains the nucleic acid molecule as described in claim 4 or 5.

7. A cell, characterized in that: The cell contains either the nucleic acid molecule of claim 4 or the expression vector of claim 6.

8. An immunoassay kit, characterized in that: The kit contains the anti-human CD61 antibody as described in any one of claims 1-3.

9. The anti-human CD61 antibody according to any one of claims 1-3 is used to prepare a kit for detecting human CD61.

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