Monoclonal antibody against novel coronavirus np protein, and application and product thereof

Abstract

This invention provides a monoclonal antibody against the NP protein of the novel coronavirus, its applications, and products, relating to the field of biotechnology. The monoclonal antibody against the NP protein of the novel coronavirus provided by this invention exhibits excellent specific binding ability to both the NP protein of the novel coronavirus and the mutant strain (Omicron) NP protein, superior to existing antibodies against the NP protein of the novel coronavirus, and can be used for the detection of the NP protein of the novel coronavirus.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a monoclonal antibody against the NP protein of the novel coronavirus, its applications, and products. Background Technology

[0002] SARS-CoV-2 belongs to the family Coronaviridae, genus Coronavirus, and is a type of enveloped, single-stranded, positive-sense RNA virus. Like Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV), it is a zoonotic pathogen originating from animals. Although coronaviruses are typically associated with acute respiratory infections in humans, their ability to infect multiple host species makes them complex pathogens. The novel coronavirus continues to evolve, producing numerous viral variants, which severely impacts the effectiveness of monoclonal antibody treatments for COVID-19.

[0003] Therefore, distinguishing different variants and identifying the prevalent strain in the current population is of great guiding significance for the selection of monoclonal antibody treatments for COVID-19. Currently known detection methods include real-time quantitative PCR, serum antibody detection, and antigen detection. Among these, antigen detection, which uses monoclonal antibodies to detect the corresponding NP antigen targets of SARS-CoV-2, is an accurate, rapid, simple, and easy-to-use diagnostic method. Monoclonal antibodies, as the core raw material of antigen detection, play a crucial role.

[0004] In view of this, the present invention is hereby proposed. Summary of the Invention

[0005] The first objective of this invention is to provide a monoclonal antibody against the NP protein of the novel coronavirus, in order to solve at least one of the aforementioned problems.

[0006] The second objective of this invention is biological materials.

[0007] The third objective of this invention is the application of the above-mentioned monoclonal antibody against the novel coronavirus NP protein in the preparation of novel coronavirus NP protein detection products.

[0008] The fourth objective of this invention is to provide a novel coronavirus NP protein marker.

[0009] The fifth objective of this invention is a kit for detecting the NP protein of the novel coronavirus.

[0010] To achieve the above objectives, the following solutions are proposed:

[0011] In a first aspect, the present invention provides a monoclonal antibody against the novel coronavirus NP protein, wherein the variable region of the monoclonal antibody against the novel coronavirus NP protein comprises: a complementarity-determining region CDR1-VH having the amino acid sequence shown in SEQ ID NO.1, a complementarity-determining region CDR2-VH having the amino acid sequence shown in SEQ ID NO.2, a complementarity-determining region CDR3-VH having the amino acid sequence shown in SEQ ID NO.3, a complementarity-determining region CDR1-VL having the amino acid sequence shown in SEQ ID NO.4, a complementarity-determining region CDR2-VL having the amino acid sequence shown in SEQ ID NO.5, and a complementarity-determining region CDR3-VL having the amino acid sequence shown in SEQ ID NO.6.

[0012] As a further technical solution, the variable region includes a heavy chain variable region VH having an amino acid sequence as shown in SEQ ID NO.7.

[0013] As a further technical solution, the variable region includes a light chain variable region VL having an amino acid sequence as shown in SEQ ID NO.8.

[0014] As a further technical solution, the monoclonal antibody against the novel coronavirus NP protein includes an IgG antibody.

[0015] Secondly, the present invention provides a biomaterial selected from any one of the following:

[0016] a. Nucleotides, said nucleotides comprising a nucleotide sequence encoding a monoclonal antibody against the novel coronavirus NP protein;

[0017] b. A carrier carrying the nucleotides in a;

[0018] c. A cell carrying the nucleotides in a, or containing the vector in b, or expressing a monoclonal antibody against the novel coronavirus NP protein.

[0019] Thirdly, the present invention provides the application of the above-mentioned monoclonal antibody against the novel coronavirus NP protein in the preparation of novel coronavirus NP protein detection products.

[0020] Fourthly, the present invention provides a novel coronavirus NP protein marker, comprising a monoclonal antibody against the novel coronavirus NP protein and the marker;

[0021] The monoclonal antibody against the novel coronavirus NP protein was conjugated with a marker.

[0022] As a further technical solution, the markers include enzymes, fluorescent molecular markers, fluorescent microspheres, colored microspheres, colloidal gold, biotin, or streptavidin.

[0023] Fifthly, the present invention provides a kit for detecting the NP protein of the novel coronavirus, the kit comprising the monoclonal antibody against the NP protein of the novel coronavirus or the NP protein marker of the novel coronavirus.

[0024] As a further technical solution, the kit includes an immunochromatographic detection kit, an ELISA detection kit, an immunomagnetic microparticle detection kit, an immunofluorescence detection kit, or an immunoblotting detection kit.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The monoclonal antibody against the novel coronavirus NP protein provided by this invention has good specific binding ability to the novel coronavirus NP protein and the mutant strain (Omicron) NP protein, which is superior to existing anti-novel coronavirus NP protein antibodies and can be used for the detection of the novel coronavirus NP protein. Attached Figure Description

[0027] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0028] Figure 1 The protein electrophoresis diagram provided for the example. Detailed Implementation

[0029] The embodiments and examples of the present invention will be described in detail below. However, those skilled in the art will understand that the following embodiments and examples are for illustrative purposes only and should not be considered as limiting the scope of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Unless otherwise specified, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0030] The "variable region" or "variable domain" of an antibody refers to the domain at the amino terminus of the antibody's heavy or light chain that recognizes and binds to antigens. The composition and arrangement of the amino acids in this region determine the antibody's specificity in recognizing antigens. The heavy chain variable region can be called "VH," and the light chain variable region can be called "VL." These domains are usually the most variable parts of the antibody and contain antigen-binding sites. The variable regions of both the heavy and light chains consist of three complementarity-determining regions (CDRs) (also known as hypervariable regions) connected by four framework regions (FRs). The CDRs in each chain are held tightly together by the FRs to form the variable region. Typically, the VL / VH variable regions of the heavy and light chains are obtained by connecting the following numbered CDRs with FRs in the following combination: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0031] In a first aspect, the present invention provides a monoclonal antibody against the novel coronavirus NP protein, wherein the variable region of the monoclonal antibody against the novel coronavirus NP protein comprises: a complementarity-determining region CDR1-VH having the amino acid sequence shown in SEQ ID NO.1, a complementarity-determining region CDR2-VH having the amino acid sequence shown in SEQ ID NO.2, a complementarity-determining region CDR3-VH having the amino acid sequence shown in SEQ ID NO.3, a complementarity-determining region CDR1-VL having the amino acid sequence shown in SEQ ID NO.4, a complementarity-determining region CDR2-VL having the amino acid sequence shown in SEQ ID NO.5, and a complementarity-determining region CDR3-VL having the amino acid sequence shown in SEQ ID NO.6.

[0032] The amino acid sequences of SEQ ID NO.1-SEQ ID NO.6 are shown in Table 1.

[0033] Table 1 Complementarity Determinant Region Sequence

[0034] High-variable area sequence serial number CDR1-VH SYAFS SEQ ID NO.1 CDR2-VH GIIPIFGTANYAQKFQG SEQ ID NO.2 CDR3-VH DLYWGSAYHYYHMDV SEQ ID NO.3 CDR1-VL GGNNIGSKSVH SEQ ID NO.4 CDR2-VL YDSDRPS SEQ ID NO.5 CDR3-VL QVWDSSSDHVV SEQ ID NO.6

[0035] In some alternative embodiments, the variable region includes a heavy chain variable region VH having an amino acid sequence as shown in SEQ ID NO.7.

[0036] QVQLVQSGAEVKKPGSSVKVSCKASGGTFRSYAFSWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDLYWGSAYHYYHMDVWGQGTTVTVSS (SEQ ID NO. 7).

[0037] In some alternative embodiments, the variable region includes a light chain variable region VL having an amino acid sequence as shown in SEQ ID NO. 8.

[0038] SYVLTQPPSVSVAPGKTARISCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDHVVFGGGTKLTVL (SEQ ID NO. 8).

[0039] In some alternative implementations, the monoclonal antibody against the novel coronavirus NP protein includes, but is not limited to, IgG antibodies.

[0040] Secondly, the present invention provides a biomaterial selected from any one of the following:

[0041] a. Nucleotides, said nucleotides comprising a nucleotide sequence encoding a monoclonal antibody against the novel coronavirus NP protein;

[0042] b. A carrier carrying the nucleotides in a;

[0043] c. A cell carrying the nucleotides in a, or containing the vector in b, or expressing a monoclonal antibody against the novel coronavirus NP protein.

[0044] Thirdly, the present invention provides the application of the above-mentioned monoclonal antibody against the novel coronavirus NP protein in the preparation of novel coronavirus NP protein detection products.

[0045] The monoclonal antibody against the novel coronavirus NP protein provided by this invention can specifically recognize the novel coronavirus and therefore can be used for the detection of the novel coronavirus.

[0046] Fourthly, the present invention provides a novel coronavirus NP protein marker, comprising a monoclonal antibody against the novel coronavirus NP protein and the marker;

[0047] The monoclonal antibody against the novel coronavirus NP protein was conjugated with a marker.

[0048] This marker can be used for specific labeling of the novel coronavirus.

[0049] In some alternative embodiments, the label includes enzymes, fluorescent molecular labels, fluorescent microspheres, colored microspheres, colloidal gold, biotin, or streptavidin.

[0050] Fifthly, the present invention provides a kit for detecting the NP protein of the novel coronavirus, the kit comprising the monoclonal antibody against the NP protein of the novel coronavirus or the NP protein marker of the novel coronavirus.

[0051] In some alternative implementations, the kit includes an immunochromatographic assay kit, an ELISA kit, an immunomagnetic microparticle assay kit, an immunofluorescence assay kit, or an immunoblotting assay kit.

[0052] The present invention will be further illustrated below with specific embodiments. However, it should be understood that these embodiments are merely for the purpose of more detailed illustration and should not be construed as limiting the present invention in any way.

[0053] Example 1: Screening of SARS-CoV-2NP monoclonal antibodies

[0054] 1. Obtaining recombinant antigens

[0055] 1.1 Construction of prokaryotic expression plasmids

[0056] Based on data on nucleocapsidphosphoprotein (NP protein, MN908947.3) in the novel coronavirus (SARS-CoV-2) database published by NCBI, Shanghai Ruier Biotechnology Co., Ltd. commercially synthesized pET28a-N (1-419, P13L, D63G, R203K, G204R, D377Y) and pET30a-N (63-413) plasmids.

[0057] 1.2 NP protein expression and purification

[0058] The synthesized plasmids were transformed into E. coli BL21(DE3) competent cells. Two SARS-CoV-2 NP proteins, NP-11 (pET28a-N(1-419,P13L,D63G,R203K,G204R,D377Y,S413R)) and NP-12 (pET30a-N(63-413)), were obtained using conventional gene recombination expression methods. The proteoglycan image is shown below. Figure 1 (From left to right in the figure: Marker, NP-11 and NP-12) As shown.

[0059] 2. Immunized animals

[0060] 2.1 Reagent Preparation

[0061] The NP-11 protein was diluted at a concentration of 400 ug / ml in autoclaved and filtered through a 0.22 μm membrane in physiological saline and gently mixed using a vortex mixer.

[0062] 2.2 The immunization schedule is as follows:

[0063] Table 2. Mouse immunization procedure

[0064]

[0065] Five healthy female Balb / c mice aged 7-8 weeks were used. The NP-11 recombinant antigen was thoroughly mixed with an equal amount of Freund's complete adjuvant, and the mice were immunized according to the above-described immunization procedure. After the fourth immunization, blood was collected by tail amputation, and the titer was determined by indirect ELISA. Mice with the best immunization were selected, and RNA was obtained from their spleen and peripheral blood mononuclear cells (PBMCs).

[0066] If the titer is low after the fourth immunization, immunization is continued. After the fifth immunization, blood is collected from the tail and the titer is determined by indirect ELISA. At the same time, mouse spleen and peripheral blood mononuclear cells (PBMCs) are obtained for RNA extraction.

[0067] 3 Antibody Screening

[0068] 3.1 Isolation and screening of spleen B cells

[0069] (1) Obtain fresh, sterile, rinsed and immunocompetent mouse spleen tissue;

[0070] (2) A sterile mouse spleen single-cell suspension was obtained by grinding and filtering with nylon mesh;

[0071] (3) Isolate lymphocytes from the obtained single-cell suspension;

[0072] (4) Take 1 mL of lymphocytes (approximately 1*10) 7 Mix magnetic beads (100 μg / ml) with 100 μg of SARS-CoV-2NP-12 protein-conjugated beads until homogeneous, and incubate at 4°C with slow rotation for 30 min.

[0073] (5) Discard any cells that have not bound to the magnetic beads and rinse the magnetic beads three times with PBS containing 2% FBA.

[0074] (6) Obtain a specific B cell suspension targeting NP-12;

[0075] 3.2 Isolation and screening of peripheral blood B cells

[0076] (1) PBMC cells in mouse blood samples were separated by density gradient centrifugation (using polysucrose-diatrizoate meglumine separation solution);

[0077] (2) Take a portion of PBMC cells and freeze them in liquid nitrogen;

[0078] (3) Take 1 mL of PBMC cells (approximately 1*10) 7 Mix magnetic beads (100 μg / ml) with 100 μg of SARS-CoV-2NP-12 protein-conjugated beads until homogeneous, and incubate at 4°C with slow rotation for 30 min.

[0079] (4) Discard any cells that have not bound to the magnetic beads and rinse the magnetic beads three times with PBS containing 2% FBA.

[0080] (5) Obtain a specific B cell suspension targeting NP-12;

[0081] 3.3 Single B-cell sequencing

[0082] Using cells from the two cell sources mentioned above, single B cell libraries were constructed using BCR sequencing. Bioinformatics methods were used to identify common clones in the two libraries, and candidate antibody sequences were selected through analysis.

[0083] 3.4 Antibody Expression

[0084] An expression vector was constructed based on the antibody sequence. The candidate antibodies were transiently expressed in Expi293F cells. The expressed antibodies were purified using rProtein A according to the manufacturer's instructions. Finally, ELISA was used to screen for antibodies with stable high affinity for NP-12 protein. After several rounds of screening, three candidate antibodies with high affinity for NP-12 were obtained. The comparison results with commercially available NP monoclonal antibodies are as follows:

[0085] Table 3 ELISA Screening Results

[0086]

[0087] The experimental data above show that candidate antibody 1 has a similar titer to the purchased NP antibody, candidate antibody 3 is inferior to the purchased NP antibody, and candidate antibody 2 is superior to the other three antibodies. It is named NP-12mAb, and its VH / VL gene amino acid sequence is as follows:

[0088] Heavy chain variable region sequence:

[0089] QVQLVQSGAEVKKPGSSVKVSCKASGGTFRSYAFSWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDLYWGSAYHYYHMDVWGQGTTVTVSS (SEQ ID NO. 7).

[0090] Light chain variable region sequence:

[0091] SYVLTQPPSVSVAPGKTARISCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDHVVFGGGTKLTVL (SEQ ID NO. 8).

[0092] Example 2: Conformity Detection of SARS-CoV-2 NP Monoclonal Antibody (NP-12mAb)

[0093] This experiment performed conformity testing on NP-12mAb. After screening, it was determined that NP-12mAb should be labeled with colloidal gold. Purchased antibody 1 (an antibody for detecting SARS-CoV-2 NP antigen) was coated on an NC membrane to prepare a colloidal gold reagent kit. The kit was then compared with corresponding SARS-CoV-2 antigen detection kits on the market. The specific experiments are as follows:

[0094] 1. Experimental combination

[0095] Table 4 Reagent Kit

[0096]

[0097] Note: Coating material dosage: 1.0 mg / ml * 0.9 μl / cm; Labeling material dosage: 0.3% * 2.0 μl / cm;

[0098] 2. Experimental Materials, Methods and Results

[0099] 2.1 Experimental Procedure

[0100] (1) Experimental scheme

[0101] The NP-12mAb antibody was prepared into a label pad according to the dosage shown in 1, and assembled into a large card according to the experimental group combination in 1. The card was then cut into reagent strips for testing.

[0102] (2) Test method

[0103] 1) Buffer: The buffer is PBS. Use a pipette to directly load 100ul into the sample well of the reagent plate. Read the results after 15 minutes and 30 minutes.

[0104] 2) Positive control: Use a pipette to directly load 100 μL of the positive control to be tested into the sample well of the reagent plate, and read the results after 15 minutes. Each positive control should be tested twice.

[0105] a: NP-11: Diluted with PBS to 500 pg / ml and 125 pg / ml;

[0106] b: CDC vaccine: Dilute with PBS to 1:5w; dilute with PBS to 1:10w;

[0107] c: Recombinant NP protein of mutant strain: wild-type strain-Ag6;

[0108] Delta-Ag52;

[0109] Alfa-Ag37;

[0110] Omecron BA.1-Ag150;

[0111] Omecron BA.2-Ag155;

[0112] Omecron BA.4-Ag159.

[0113] 3) Virus culture (this part is completed by the CDC testing department):

[0114] a: Wild-type strain-P1 (test concentration: CT value 26.27 / 35.16 / NA-35.16 diluted 10 times);

[0115] b: Delta-P2 (test concentration: CT value 28.37 / 36.95 / NA-36.95 diluted 10 times);

[0116] c: BA.1-P3 (test concentration: CT value 24.52 / 30.49 / 36.82);

[0117] d: BA.2.3-P4 (test concentration: CT value 24.57 / 30.71 / 35.4).

[0118] 4) Interference samples: Use a pipette to directly load 100 μL of the interference sample to be tested into the sample well of the reagent plate, and read the results after 15 minutes and 30 minutes;

[0119] a: HAMA: SB124452, diluted 1:5 with PBS buffer;

[0120] b: HAMA: 3PH490, diluted 1:5 with PBS buffer;

[0121] c:HAMA:3PH498.

[0122] 5) Negative samples: Prepare negative samples (PBS / nasal secretion samples) according to the instructions, and interpret the results at 15 minutes and 30 minutes.

[0123] 6) False positive serum sample: 20ul serum (selected false positive serum sample), add 80ul PBS, and interpret the results after 15 minutes.

[0124] 7) Clinical positive samples: Collect nasal secretion samples from different infection stages according to the instructions, prepare them into clinical positive samples according to the experimental steps in the instructions, and interpret the results in 15 minutes.

[0125] (3) Other

[0126] a: Color chart;

[0127] b: Instrument: ACG1000-A002.

[0128] (4) Experimental Results

[0129] Table 5 Positive control test results - machine readings

[0130]

[0131] Note: *0 indicates a negative test result. Under the same membrane background conditions, the higher the machine value, the better the sensitivity.

[0132] The results above show that the kit composed of NP-12mAb has higher sensitivity than the purchased kit when testing NP-11 recombinant protein; when testing CDC vaccine, both kits have relatively low overall sensitivity, with the purchased kit being slightly better than the kit composed of NP-12mAb.

[0133] Table 6. Positive control (mutant NP protein) test results - machine reading

[0134]

[0135] The results above show that the kit composed of NP-12mAb has higher sensitivity than the purchased kit when testing NP recombinant proteins from mutant strains.

[0136] Table 7 Virus Cultures—Disease Control Testing

[0137]

[0138] Note: "+" indicates a positive result, and "-" indicates a negative result; the intensity of the T line is determined by manually reading the test strip against the standard color chart, and a value of 3 or below indicates a negative result;

[0139] The results above show that the kit composed of NP-12mAb has higher sensitivity than commercially available kits when testing viral cultures.

[0140] Table 8. Results of NP protein testing in different mutant strains of Omicron

[0141]

[0142] The results above show that the kit composed of NP-12mAb has higher sensitivity than the commercially available kit at a concentration of 100 pg / ml when testing NP recombinant protein mutants; and slightly better sensitivity than the commercially available kit at a concentration of 25 pg / ml.

[0143] Table 9. Test Results of Interference Specimens

[0144]

[0145] Note: *0 indicates a negative test result. Under the same membrane background, the higher the machine value, the higher the risk of a false positive.

[0146] The results above show that the kit composed of NP-12mAb produced weak false positives when testing HM452 and HM490 interference samples; the purchased kit produced significant false positives when testing HM452 and HM490 interference samples, and weak false positives when testing HM498 interference samples.

[0147] Table 10 Negative Sample Test Results

[0148]

[0149] The results above show that neither the kit composed of NP-12mAb nor the purchased kit showed obvious false positives.

[0150] Table 11 Clinical Sample Test Results

[0151]

[0152]

[0153] Note: The strong positive sample used in Y6 is a different strong positive sample from Y3.

[0154] The results above show that the NP-12mAb kit performs comparably to commercially available kits when testing clinically positive samples; and outperforms commercially available kits when testing false-positive serum samples.

[0155] 3. Experimental Conclusions

[0156] The above experimental results demonstrate that the colloidal gold reagent kit prepared by combining NP-12mAb with purchased antibody 1 is effective.

[0157] (1) Positive control:

[0158] a. When testing NP-11 quality control samples, its performance is superior to that of purchased reagent kits;

[0159] b. When testing the CDC vaccine, its performance was inferior to that of purchased test kits;

[0160] c. When testing recombinant proteins from partial mutant strains, the performance was significantly better than that of commercially available kits;

[0161] d. When testing partial viral cultures, it outperformed commercially available kits;

[0162] (2) Interference samples:

[0163] When testing interfering samples, its performance was significantly better than that of purchased reagent kits;

[0164] (3) Negative samples:

[0165] When testing negative samples, its performance is comparable to that of commercially available kits;

[0166] (4) Clinical samples:

[0167] When testing clinical positive samples, its performance is comparable to that of commercially available kits;

[0168] When testing false positive serum samples, it outperforms commercially available kits.

[0169] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A monoclonal antibody against the NP protein of the novel coronavirus, characterized in that, The variable region of the monoclonal antibody against the novel coronavirus NP protein includes: the complementarity-determining region CDR1-VH of the amino acid sequence shown in SEQ ID NO.1, the complementarity-determining region CDR2-VH of the amino acid sequence shown in SEQ ID NO.2, the complementarity-determining region CDR3-VH of the amino acid sequence shown in SEQ ID NO.3, the complementarity-determining region CDR1-VL of the amino acid sequence shown in SEQ ID NO.4, the complementarity-determining region CDR2-VL of the amino acid sequence shown in SEQ ID NO.5, and the complementarity-determining region CDR3-VL of the amino acid sequence shown in SEQ ID NO.

6.

2. The monoclonal antibody against the novel coronavirus NP protein according to claim 1, characterized in that, The variable region includes the heavy chain variable region VH of the amino acid sequence shown in SEQ ID NO.

7.

3. The monoclonal antibody against the novel coronavirus NP protein according to claim 1, characterized in that, The variable region includes the light chain variable region VL of the amino acid sequence shown in SEQ ID NO.

8.

4. The monoclonal antibody against the novel coronavirus NP protein according to claim 1, characterized in that, The monoclonal antibody against the novel coronavirus NP protein includes an IgG antibody.

5. A biomaterial, characterized in that, The biomaterial is selected from any one of ac: a. Nucleic acid, wherein the nucleic acid is a nucleic acid sequence encoding a monoclonal antibody against the novel coronavirus NP protein according to any one of claims 1-4; b. A vector carrying the nucleic acid from a; c. A cell carrying the nucleic acid of a, or containing the vector of b, or expressing a monoclonal antibody against the novel coronavirus NP protein as described in any one of claims 1 to 4.

6. The use of the monoclonal antibody against the novel coronavirus NP protein as described in any one of claims 1-4 in the preparation of a novel coronavirus NP protein detection product.

7. A novel coronavirus NP protein marker, characterized in that, Includes the monoclonal antibody and marker against the novel coronavirus NP protein as described in any one of claims 1-4; The monoclonal antibody against the novel coronavirus NP protein was conjugated with a marker.

8. The novel coronavirus NP protein marker according to claim 7, characterized in that, The label is selected from enzymes, fluorescent molecular labels, fluorescent microspheres, colored microspheres, colloidal gold, biotin, or streptavidin.

9. A kit for detecting NP protein of the novel coronavirus, characterized in that, The kit comprises a monoclonal antibody against the novel coronavirus NP protein as described in any one of claims 1-4 or a novel coronavirus NP protein marker as described in claim 7 or 8.

10. The reagent kit according to claim 9, characterized in that, The kit is selected from immunochromatographic assay kits, ELISA kits, immunomagnetic microparticle assay kits, immunofluorescence assay kits, or immunoblotting assay kits.

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