Anti-giant panda CDV monoclonal antibody, hybridoma cell line and use thereof

Abstract

The present invention discloses an anti-giant panda CDV monoclonal antibody, hybridoma cell line, and uses thereof, belonging to the field of biotechnology. The amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 4. Using cell fusion technology, the present invention obtained the hybridoma cell line 4A3 that secretes anti-giant panda CDV monoclonal antibodies. The cell line culture supernatant was further collected and purified to obtain a highly specific monoclonal antibody.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to an anti-giant panda CDV monoclonal antibody, a hybridoma cell line and uses thereof. Background Art

[0002] Canine distemper (CD) is a highly contagious and fatal disease of a variety of carnivores caused by the canine distemper virus (CDV). CDV particles are pleomorphic (usually round) and belong to the Paramyxoviridae family and the Morbillivirus genus. They are enveloped, single-stranded, negative-sense, nonsegmented RNA viruses with diameters ranging from 100 to 250 nm. CDV can infect a wide range of animals. Due to the expansion of human activities and wildlife habitats, its host range has expanded. In addition to canids, CDV can also infect a variety of species, including mustelids, hyenas, raccoons, and pandas, including giant pandas, tigers, lions, red pandas, lynxes, bears, and wolves. In recent years, CDV's host range has expanded to primates such as macaques and cynomolgus macaques, as well as rodents. Between 2014 and 2015, six giant pandas were found to be infected with CDV, resulting in the deaths of five of them.

[0003] CDV, similar to other paramyxoviruses, contains six structural proteins: matrix protein (M), hemagglutinin protein (H), fusion protein (F), phosphoprotein (P), large protein (L), and nucleocapsid protein (N). These proteins are arranged in the order of leader sequence (3'), NPMFHL (NPMFHL), and trailer sequence (5'). The P protein gene also contains two nonstructural protein genes, V and C.

[0004] The CDV H protein is a structural protein essential for the virus to invade the host and is one of the main antigens used by the virus to stimulate the body to produce neutralizing antibodies. The CDV H protein is approximately 78 kDa in size and is located on the particle surface as a tetramer. It can bind to host-specific receptors, thereby mediating viral infection of cells. The H protein is a type II glycoprotein and contains three regions: an N-terminal cytoplasmic tail, a transmembrane region, and a C-terminal extracellular domain. The cytoplasmic tail contains the transmembrane signal region and anchoring site, while the C-terminus forms the "head" of the H protein that can bind to the receptor. The H protein primarily performs two functions: first, it enables the virus to bind to cell surface receptors; second, it triggers the cell fusion process guided by the F protein. These characteristics make the H protein an ideal structural protein for studying the polymorphism of CDV strains and molecular epidemiology.

[0005] According to the latest statistics, the captive giant panda population in China currently numbers approximately 630 individuals. As the density of captive populations increases, the spread and mutation of viruses among animals accelerates, posing a serious threat to the health and stability of the population. There are currently no specific preventive or therapeutic medications for canine distemper, a high-risk viral infectious disease threatening the health of giant pandas. Vaccines used to immunize giant pandas against canine distemper are mostly designed for dogs, but due to species differences, they lack effective antibodies, have a short protective period, and carry the potential risk of inducing infection.

[0006] Currently, clinical treatment for canine distemper primarily involves symptomatic treatment with hyperimmune serum combined with antiviral drugs. Studies on antibodies targeting panda-derived CDV viruses are limited. Studies have found that CDV monoclonal antibodies are more stable and effective than hyperimmune serum. Therefore, the hope is to develop monoclonal antibodies against panda-derived CDV to better protect the giant panda population. Summary of the Invention

[0007] In view of the above-mentioned deficiencies in the prior art, the present invention provides an anti-giant panda CDV monoclonal antibody, a hybridoma cell line and uses thereof, which can effectively apply the obtained monoclonal antibody to the treatment of canine distemper derived from giant pandas.

[0008] To achieve the above-mentioned purpose, the technical solution adopted by the present invention to solve the technical problem is:

[0009] The purpose of the present invention is to provide an anti-giant panda CDV monoclonal antibody, the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO.2, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO.4.

[0010] Furthermore, the subtype of the monoclonal antibody (4A3 antibody) is mouse IgG1*03 or IgG1*04 subtype.

[0011] Furthermore, the nucleic acid sequence encoding the amino acids of the heavy chain variable region of the anti-giant panda CDV monoclonal antibody is shown as SEQ ID NO.1; the nucleic acid sequence encoding the amino acids of the light chain variable region of the anti-giant panda CDV monoclonal antibody is shown as SEQ ID NO.3.

[0012] Another object of the present invention is to provide an antibody composition comprising the above-mentioned anti-giant panda CDV monoclonal antibody, and a pharmaceutically acceptable carrier, buffer and excipient.

[0013] Another object of the present invention is to provide a kit comprising the above-mentioned anti-giant panda CDV monoclonal antibody or antibody composition.

[0014] Another object of the present invention is to provide a hybridoma cell line 4A3 that secretes the above-mentioned anti-giant panda CDV monoclonal antibody, which was deposited in the China Center for Type Culture Collection, Wuhan University, Wuhan, China on April 23, 2025, with a deposit number of CCTCC NO: C2025133.

[0015] Another object of the present invention is to provide the use of the above-mentioned anti-giant panda CDV monoclonal antibody, antibody composition or kit in basic medical research for non-diagnostic / therapeutic purposes.

[0016] Furthermore, basic medical research includes Western Blot, immunohistochemistry or flow cytometry analysis.

[0017] Another object of the present invention is to provide the use of the above-mentioned anti-giant panda CDV monoclonal antibody or antibody composition in the preparation of a preparation for detecting giant panda CDV.

[0018] Beneficial effects of the present invention:

[0019] Using cell fusion technology, the present invention generated a hybridoma cell line 4A3 that secretes a monoclonal antibody against giant panda CDV. The cell culture supernatant was then collected and purified to obtain a highly specific monoclonal antibody (4A3). Western blot analysis and ELISA assays revealed that the monoclonal antibody had excellent binding activity with the CDV H protein, providing a reference for the subsequent preparation of chimeric antibodies against canine distemper in giant pandas. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The figure shows the expression results of the purified monoclonal antibody (4A3) analyzed by SDS-PAGE. Band M: SDS-PAGE Protein Marker; Band 4A3: purified monoclonal antibody band.

[0021] Figure 2 Figure 1 is a diagram of sequencing results; Figure A is an agarose gel electrophoresis diagram of the amplified protein by PCR using mIGHG primers; Band M in the figure: SDS-PAGE Protein Marker; Band 4A3: 4A3 amplified by PCR using mIGHG primers; Figure B is the sequencing result diagram after recovering the target band 4A3;

[0022] Figure 3 The figure shows the Western blot identification result of the monoclonal antibody binding to CDV-H protein; wherein, band M: SDS-PAGE Protein Marker; band CDV-H: band of 4A3 binding to CDV-H protein;

[0023] Figure 4 EC50 is the half-maximal effective concentration (EC) of monoclonal antibodies detected by ELISA. 50 Result graph of the values. DETAILED DESCRIPTION

[0024] The specific embodiments of the present invention are described below to facilitate understanding of the present invention by those skilled in the art. However, it should be clear that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the appended claims, these changes are obvious, and all inventions and creations utilizing the concepts of the present invention are protected.

[0025] Example 1 Establishment of Monoclonal Antibody Hybridoma Cell Line

[0026] 1. Expression and purification of CDV H protein

[0027] Based on the original eukaryotic expressed CDV H protein, the purified recombinant protein was obtained by Pujian Biotechnology (Wuhan) Co., Ltd. through plasmid mini-extraction, plasmid transformation, and plasmid purification. The concentration of the protein was determined to be 0.46 mg / mL using a BCA protein quantification kit (product of Thermo Fisher Scientific) and stored at -80°C until use.

[0028] 2. Animal immunization

[0029] The purified eukaryotically expressed CDV H protein (0.5 mg / mL) was emulsified with an equal volume of QuickAntibody-Mouse3W aqueous adjuvant (Biolong) and injected intramuscularly into 6-8 week old female BALB / c mice at 100 μL / mouse. Thereafter, it was emulsified with an equal volume of Freund's incomplete adjuvant (Sigma) every 14 days and immunized three more times. Seven days after the second, third, and fourth immunizations, the blood of the mice was collected and the serum was separated. The antibody titer was detected by indirect ELISA test and neutralization test. Seven days after the fourth immunization, cell fusion was performed, blood was collected and serum was separated, and the titer was detected. The serum antibody titer of the five mice (413, 414, 415, 416, 417) raised reached a maximum of 1:5120000 (2 9 ×10 4 ), the serum neutralization titer was as high as 1:2048 (2 11 ).

[0030] 3. Cell fusion

[0031] Mouse 414 cells, which had the highest serum antibody and neutralization titers, were selected for cell fusion. Using the PEG cell fusion method, well-grown SP2 / 0 myeloma cells (laboratory-preserved) were mixed with spleen cells from immunized BALB / c mice at a ratio of 1:5. The mixture was thoroughly mixed and centrifuged at 1000 rpm for 7 minutes. The supernatant was discarded. Using a 1 mL pipette, 1 mL of ClonaCell™-HY PEG (STEMCELL, Canada) was slowly added dropwise to the cells over 1 minute. The cells were gently stirred with the pipette tip for 1 minute. 4 mL of incomplete RPMI 1640 medium (Gibco, USA) without fetal bovine serum was added to the fusion mixture, and stirring was continued for 4 minutes. Then, 10 mL of incomplete RPMI 1640 medium without fetal bovine serum was slowly added to the fusion mixture. The mixture was incubated in a 37°C water bath for 15 minutes. Slowly add 30 mL of RPMI 1640 complete medium containing 10% fetal calf serum (FCS) (Gibco, USA) and 1% penicillin-streptomycin mixture (100×) (Gibco, USA), centrifuge at 1000 rpm for 7 minutes, and discard the supernatant. Slowly add 40 mL of RPMI 1640 complete medium, centrifuge at 1000 rpm for 7 minutes, and discard the supernatant. Slowly resuspend in 10 mL of RPMI 1640 complete medium. Transfer the cell suspension to a cell flask containing 20 mL of RPMI 1640 complete medium. Incubate the cell flask in a cell incubator at 37°C and 5% CO2 for 16-24 hours. Bring ClonaCell™-HY Medium D (Canadian STEMCELL Company) to room temperature one day in advance and shake the medium D vigorously to mix it thoroughly. Transfer the fused cell suspension from the T75cm 2Transfer the cells from the flask to a 50 mL conical tube, centrifuge at 1000 rpm for 7 minutes, and discard the supernatant. Resuspend the cells in RPMI 1640 complete medium to a total volume of 6 mL. Transfer the cell suspension directly to a bottle containing 60 mL of medium D. Gently invert the bottle several times to mix thoroughly. Incubate at room temperature for 15 minutes, allowing bubbles to rise to the top. Using a 20 mL syringe and a blunt-end needle, slowly add 1 mL of cell suspension in medium D to a 6-well plate, taking care to avoid generating bubbles during the process. Tilt each plate to evenly distribute the medium on the bottom of the plate. Incubate in a 37°C, 5% CO2 cell incubator for 10-14 days. Observe the cell status under a microscope every day after 7 days of cell fusion.

[0032] 4. Screening of hybridoma cell lines

[0033] Using a pipette and pipette tip, aspirate hybridoma cells from a 6-well plate under a microscope. Transfer a single hybridoma cell to a 96-well cell culture plate containing HT medium (Gibco, USA). Mix the cells in each well by pipetting, then incubate the 96-well plate in a 37°C, 5% CO2 cell culture incubator for 3-4 days.

[0034] Dilute the protein to 2 μg / mL with coating solution, add 100 μL to each well of the ELISA plate, incubate at 4°C overnight, and wash twice with PBST, 5 min / time. Add 300 μL of blocking solution (3% skim milk powder, diluted with PBST) to each well and incubate at 37°C for 2 hours. Discard the blocking solution, add 100 μL of PBST containing 1% skim milk powder to each well, add 100 μL of hybridoma cell supernatant to the first well of each column without dilution, and perform positive analysis (dilute the positive serum of the immunized mouse to OD 450 nm value close to 1.0), negative control, incubate at 37℃ for 1h. Discard the liquid in the plate and wash three times with PBST, 5min / time. Add 100μL of 1:20000 diluted HRP-labeled goat anti-mouse enzyme-labeled secondary antibody (BS12478, Bioworld, USA) to each well of the enzyme-labeled plate, place in a 37℃ incubator, and incubate for 1h. Discard the liquid in the plate and wash five times with PBST, 5min / time. Add 100μL of TMB colorimetric solution to each well and color for 10min in the dark. Add 50μL of stop solution to each well. Measure OD using an enzyme-labeled instrument 450 nm value, zeroed with blank control, P is the value of each test well, N is the OD450nm value of negative reference serum, when the OD 450 nm values ​​and OD of negative reference serum 450 The ratio of nm values ​​is ≥2.1, that is, under the premise that negative and positive controls are established, the detection wells with P / N ≥2.1 are judged as positive.

[0035] Add 50 μL / well of incomplete DMEM (Corning, USA) without fetal bovine serum to a 96-well plate (operate on ice, pre-chill DMEM). Add 50 μL / well of CDV-Onderstepoort (vaccine strain) virus solution (diluted to an MOI of 0.1, operate on ice). Add 100 μL of Vero cell suspension to the 96-well plate and incubate in a 37°C, 5% CO2 cell incubator for 48 hours. Remove the 96-well plate, discard the medium, and fix the cells with 100 μL of -20°C pre-chilled 80% acetone solution per well for 30 minutes at room temperature. Discard the cold acetone, add 300 μL of PBS to each well, and wash on a shaker at room temperature for 5 minutes. Repeat three times. Block the plate with 100 μL of 2% BSA (in PBS) per well and incubate at 37°C for 1 hour. Discard the plate contents, add 50 μL of the supernatant from a positive ELISA-tested monoclonal antibody to each well, and add three replicates of the positive and negative control wells (200-fold dilution in PBS). Incubate at 37°C for 1 hour. Discard the plate contents, add 300 μL of PBST to each well, wash on a shaker at room temperature for 5 minutes, and repeat three times. Discard the plate contents, add 1:500 dilution of Alexa 488-conjugated goat anti-mouse fluorescent secondary antibody (A0428, Shanghai Bio-Tech Biotechnology Co., Ltd.) and 50 μL of 1:500 dilution of Evans Blue stain to each well, and incubate at 37°C for 1 hour. Discard the plate contents, add 300 μL of PBST to each well, wash on a shaker at room temperature for 5 minutes, and repeat three times (protect from light). Discard the plate contents and examine the results under an upright fluorescence microscope. Wells showing green fluorescence are considered positive. Positive hybridoma cell lines were screened by indirect ELISA and immunofluorescence identification and subcloned by semi-solid culture method.

[0036] 5. Cloning of hybridoma cell lines

[0037] Resuspend the positive hybridoma cell lines, take 100 cells after cell counting, resuspend in 1 mL RPMI1640 complete medium, add to a 15 mL centrifuge tube containing 10 mL medium D, mix by inversion, let it stand for 15 minutes, transfer to a 6-well plate, 2 mL per well, and add 2 mL of sterile PBS to the remaining well. Place in a 37°C, 5% CO2 cell culture incubator for 10-14 days, pick and screen the subcloned cells, and record the wells with only a single clone growing. After 14 days, take the cell supernatant and use the indirect ELISA method to detect the ELISA titer. A total of 9 positive hybridoma cell lines 4A3 were screened.

[0038] 6. Serum-free acclimation and purification of monoclonal antibodies

[0039] Resuscitate hybridoma cell line 4A3 in 1640 complete medium (Gibco1640+10% FBS+1% P / S) and culture until the cells are in good condition. Transfer them to serum-free medium (OPM-293CD05 Medium) and continuously adapt and culture for more than three generations. When they are in good condition, culture them at 5×10 5 cells / mL were transferred into serum-free medium and cultured in a CO2 shaker at 37°C and 125 rpm / min for 5-7 days until the cell viability dropped to about 60%. The cell supernatant was collected and purified using a Protein A column to obtain the antibody. The purity was determined by SDS-PAGE. After the purified antibody was analyzed correctly by SDS-PAGE, it was stored at -80°C. The SDS-PAGE results are shown in the table. Figure 1 .

[0040] 7. Subtype Verification

[0041] RNA from the hybridoma cell line 4A3 was extracted and reverse transcribed into cDNA. PCR amplification was performed using PrimeSTAR Mix enzyme and mIGHG-CH2 primers. The amplification temperature was 98°C for 10 seconds, 55°C for 5 seconds, and 72°C for 10 seconds for 35 cycles. The PCR system was 200 μL amplification. After gel excision and recovery of the target band, sequencing was performed using IGHG_CH2 primers and alignment was performed using IGMT. PCR results ( Figure 2 Figure A in the middle shows that a 1000bp band was amplified using the mIGHG primer, which is consistent with the target size, proving that it is an IgG antibody. Sequencing results ( Figure 2 Middle panel B) shows that the antibody is of mouse IgG1*03 or IgG1*04 subtype.

[0042] 8. Cryopreservation

[0043] After the hybridoma cell line 4A3, which secretes a giant panda-derived monoclonal antibody against canine distemper virus H protein, has stabilized, the cells in the culture flask were pipetted to resuspend them in the culture medium (cells are typically suspended in the culture medium or adherent). The cells were then transferred to a 15 mL centrifuge tube. Centrifugation was performed at 1000 rpm for 5 minutes. Washes were performed twice with PBS: first, the supernatant was aspirated from the centrifuge tube, discarded, PBS was added, mixed, and centrifuged at 1000 rpm for 5 minutes. This was repeated once. Finally, the supernatant was aspirated with a pipette, and the appropriate amount of freezing solution (freezing solution = 5 mL serum + 4 mL DMEM + 1 mL DMSO) was added to the cells. The mixture was inverted and filtered for later use. Finally, 1 mL of cell suspension was added to the cryovials. Place the hybridoma cell line 4A3 in a freezing box and store it at -80°C overnight. Then, place it in liquid nitrogen for long-term storage. It was deposited in the China Type Culture Collection of Wuhan University, Wuhan, China on April 23, 2025, and its deposit number is CCTCC NO: C2025133.

[0044] Example 2 Verification of Antibody Activity of Monoclonal Antibodies

[0045] 1. Western blot identification

[0046] A 5 μg sample of panda CDV H protein was subjected to SDS-PAGE electrophoresis. After electrophoresis, the protein was transferred to a PVDF membrane. The membrane was blocked with 5% skim milk powder at room temperature for 1 hour. Then, 4A3 monoclonal antibody was diluted to 3 μg / mL in PBST containing 5% skim milk powder and incubated overnight at 4°C. The membrane was washed three times with PBST and anti-mouse IgG (H+L) fluorescent secondary antibody (1:5000 dilution) was added and incubated in the dark at room temperature for 1 hour. The membrane was washed three times with PBST and scanned and imaged using an Odyssey CLx dual-color infrared laser imaging system. The results are shown in Figure 2. Figure 3 shown.

[0047] 2. ELISA identification of EC 50

[0048] Dilute the panda CDV H protein to 2µg / mL with coating solution, coat 100 µL well at 4°C overnight. Remove the coated ELISA plate, wash three times with 0.05% PBST, and block with 2% BSA at 37°C for 1 h. Remove the blocked ELISA plate, wash three times with 0.05% PBST, add the antibody, and dilute it 8 times from 10µg / mL to the negative control in 8 steps. Incubate at 37°C for 2 h. Remove the incubated ELISA plate, wash three times with 0.05% PBST, and incubate with secondary antibody (goat anti-mouse IgG (H+L)) 1:5000 (Sigma, A4416) at 37°C for 2 h. Wash three times with 0.05% PBST, add TMB for color development for 10 min, stop with stop solution (2.5M H2SO4), and read at 450 on a microplate reader. nm The OD value and the corresponding concentration gradient were input into GraphPad Prism software to calculate the half-maximal effect concentration (EC 50 ) value, the result is as follows Figure 4 shown.

[0049] 3. Sequencing

[0050] The selected hybridoma cell line 4A3 was sent to Nanjing Detai Bioengineering Co., Ltd. for sequencing. The specific process was as follows: hybridoma cells were lysed to extract total RNA, which was then reverse-transcribed into cDNA using RACE technology. The heavy and light chain variable region sequences were then amplified by PCR. The target fragments were then ligated into a vector using ligase, and the ligated product was transformed into competent E. coli cells. Single clones were then selected for sequencing, and the sequencing results were analyzed and annotated.

[0051] The heavy chain variable region base sequence of the anti-giant panda CDV monoclonal antibody finally obtained is shown in SEQ ID NO.1, the heavy chain variable region amino acid sequence is shown in SEQ ID NO.2, the light chain variable region base sequence is shown in SEQ ID NO.3, and the light chain variable region amino acid sequence is shown in SEQ ID NO.4. The specific sequences are as follows:

[0052] CACGTCCAGCTGCAGCAATCTGGACCTGAGCTGGTGAGGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTATATCTTCATCACCTACTGGATGAACTGGGTGAAGCAGAGGCCTGGACAGGGCCTTGAGTGGATTGGACAGATTTTTCCTGCAAGTGGCAGAACTAAGTACAATGAGATGTTCGAGGGCAAGGCCACATTGACTGTAGACACATCCTCCAGCACAGTTTACATGCAGCTCAGAAGCCTGACATTTGAGGACTCTGCGGTCTATTACTGTGCAAGAAAAGGGGACTGGGACGTCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTGTCCTCA(SEQ ID NO.1)。

[0053] HVQLQQSGPELVRPGASVKLSCKASGYIFITYWMNWVKQRPGQGLEWIGQIFPASGRTKYNEMFEGKATLTVDTSSSTVYMQLRSLTFEDSAVYYCARKGDWDVFDYWGQGTTLTVSS(SEQ ID NO.2)。

[0054] GATGTTGTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCCTTGTACACAGTAATGGAAACACCTATTTACATTGGTACCTGCAGAAGCCAGGCCAGTCTCCAAAGCTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGAGTGGAGGCTGAGGATCTGGGAGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA(SEQ ID NO.3)。

[0055] DVVMTQTPLSLPVSLGDQASISCRSSQSLVHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPWTFGGGTKLEIK (SEQ ID NO. 4).

[0056] Finally, it should be noted that the above specific implementation methods are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail with reference to examples, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. An anti-giant panda CDV monoclonal antibody, characterized in that: The amino acid sequence of its heavy chain variable region is shown in SEQ ID NO.2, and the amino acid sequence of its light chain variable region is shown in SEQ ID NO.

4.

2. The anti-giant panda CDV monoclonal antibody according to claim 1, characterized in that The subtype of the monoclonal antibody is mouse IgG1*03 or IgG1*04.

3. An antibody composition, characterized in that The invention comprises the anti-giant panda CDV monoclonal antibody according to claim 1 or 2, and a pharmaceutically acceptable carrier, buffer and excipient thereof.

4. A kit, characterized in that The invention comprises the anti-giant panda CDV monoclonal antibody according to claim 1 or 2 or the antibody composition according to claim 3.

5. A hybridoma cell line 4A3 secreting the anti-giant panda CDV monoclonal antibody according to claim 1 or 2, characterized in that: It was deposited in the China Type Culture Collection of Wuhan University, Wuhan, China on April 23, 2025, and its deposit number is CCTCC NO: C2025133.

6. Use of the anti-giant panda CDV monoclonal antibody according to claim 1 or 2, the antibody composition according to claim 3, or the kit according to claim 4 in basic medical research for non-diagnostic / therapeutic purposes.

7. The use according to claim 6, characterized in that The basic medical research is Western Blot, immunohistochemistry or flow cytometry analysis.

8. Use of the anti-giant panda CDV monoclonal antibody according to claim 1 or 2 or the antibody composition according to claim 3 in the preparation of a preparation for detecting giant panda CDV.

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