A monoclonal antibody against norovirus and its application

Abstract

The present invention discloses an anti-norovirus monoclonal antibody and its application. The present invention discloses a monoclonal antibody against VP1 protein, wherein the complementary determining regions HCDR1, HCDR2, and HCDR3 of the heavy chain variable region are shown as SEQ ID NOs: 1-3, the complementary determining regions LCDR1 and LCDR3 of the light chain variable region are shown as SEQ ID NOs: 4-5, and the amino acid sequence of the complementary determining region LCDR2 of the light chain variable region is NAK. The anti-norovirus monoclonal antibody disclosed in the present invention has strong affinity and high binding activity to the norovirus type 1 VP1 protein, and is an excellent monoclonal antibody for detecting and treating norovirus.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology and relates to an anti-norovirus monoclonal antibody and an application thereof. Background Art

[0002] Norovirus (NV), also known as norovirus, Norwalk virus, or pyocyanin virus, is a viral infection that causes nonbacterial acute gastroenteritis. It is particularly prone to infecting densely populated areas with poor sanitation, such as cruise ships. Norovirus is primarily transmitted through the fecal-oral route, when food is contaminated with fecal material. In densely populated areas, it can also be transmitted through respiratory droplets.

[0003] Noroviruses are divided into genogroups I and II based on their genetic makeup. Genogroup I includes norovirus, Southampton virus, and Desert Shield virus; genogroup II includes Snow Mountain virus, Hawaii virus, and Toronto virus. Common symptoms include abdominal pain, diarrhea, vomiting, low-grade fever, and generalized muscle aches. Symptoms typically resolve after 12 to 60 hours, but viral excretion can continue for about two weeks. Severe diarrhea can lead to dehydration and shock. Summary of the Invention

[0004] In order to solve the technical problems existing in the prior art, the present invention provides the following technical solutions:

[0005] The present invention provides a monoclonal antibody against VP1 protein, wherein the heavy chain variable region of the monoclonal antibody includes complementary determining regions HCDR1, HCDR2 and HCDR3 having amino acid sequences as shown in SEQ ID NOs: 1-3, and the light chain variable region includes complementary determining regions LCDR1 and LCDR3 having amino acid sequences as shown in SEQ ID NOs: 4-5, and the amino acid sequence of the complementary determining region LCDR2 of the light chain variable region is NAK.

[0006] Furthermore, the anti-VP1 protein is an anti-norovirus type 1 VP1 protein.

[0007] Furthermore, the monoclonal antibody further includes framework regions FR1, FR2, FR3, and FR4 of the heavy chain variable region and framework regions FR1, FR2, FR3, and FR4 of the light chain variable region.

[0008] Furthermore, the framework regions FR1, FR2, FR3, and FR4 of the heavy chain variable region of the monoclonal antibody are at least 70% identical, preferably at least 75% identical, preferably at least 80% identical, preferably at least 85% identical, preferably at least 90% identical, preferably at least 95% identical, preferably at least 96% identical, preferably at least 97% identical, preferably at least 98% identical, and preferably at least 99% identical to the amino acid sequences of SEQ ID NOs: 6-9, respectively.

[0009] Furthermore, the framework regions FR1, FR2, FR3, and FR4 of the light chain variable region of the monoclonal antibody are at least 70% identical, preferably at least 75% identical, preferably at least 80% identical, preferably at least 85% identical, preferably at least 90% identical, preferably at least 95% identical, preferably at least 96% identical, preferably at least 97% identical, preferably at least 98% identical, and preferably at least 99% identical to the amino acid sequences of SEQ ID NOs: 10-13, respectively.

[0010] Furthermore, the amino acid sequences of the framework regions FR1, FR2, FR3, and FR4 of the heavy chain variable region of the monoclonal antibody are shown in SEQ ID NOs: 6-9, and the amino acid sequences of the framework regions FR1, FR2, FR3, and FR4 of the light chain variable region are shown in SEQ ID NOs: 10-13.

[0011] Furthermore, the amino acid sequence of the heavy chain variable region of the monoclonal antibody has at least 60% identity, preferably at least 65% identity, preferably at least 70% identity, preferably at least 75% identity, preferably at least 80% identity, preferably at least 85% identity, preferably at least 90% identity, preferably at least 95% identity, preferably at least 96% identity, preferably at least 97% identity, preferably at least 98% identity, preferably at least 99% identity to the amino acid sequence of SEQ ID NO: 14.

[0012] Furthermore, the amino acid sequence of the light chain variable region of the monoclonal antibody has at least 60% identity, preferably at least 65% identity, preferably at least 70% identity, preferably at least 75% identity, preferably at least 80% identity, preferably at least 85% identity, preferably at least 90% identity, preferably at least 95% identity, preferably at least 96% identity, preferably at least 97% identity, preferably at least 98% identity, preferably at least 99% identity to the amino acid sequence of SEQ ID NO: 15.

[0013] Furthermore, the amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown in SEQ ID NO: 14, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 15.

[0014] Furthermore, the CDR division of the monoclonal antibody can also be performed using the Kabat, IMGT, Chothia, AbM or Contact numbering systems.

[0015] Furthermore, the CDRs of the monoclonal antibody are divided based on the amino acid sequence of the heavy chain variable region as shown in SEQ ID NO: 14 and the amino acid sequence of the light chain variable region as shown in SEQ ID NO: 15 as the basic sequences.

[0016] As used herein, the terms "sequence identity" and "identity" are synonymous and refer to the "percentage of sequence identity" or "percentage of identity" between two polynucleotides, i.e., the number of identical matching positions shared by the sequences within the comparison window, taking into account additions or deletions (i.e., gaps) that must be introduced for optimal alignment of the two sequences. A matching position is any position where the same nucleotide is present in both the target sequence and the reference sequence. Since a gap is not a nucleotide, a gap present in the target sequence is not counted. Similarly, since the target sequence nucleotides are counted and the nucleotides from the reference sequence are not counted, the gap present in the reference sequence is not counted. At least 60% sequence identity includes a continuous segment having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity over the full length of the sequence. "Percentage of sequence identity" can be calculated by comparing the two best aligned sequences in the window, determining the number of positions at which the same nucleic acid base (e.g., A, T, C, G, I) or the same amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys, and Met) appears in the two sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window (i.e., the window size), and multiplying the result by 100 to obtain the percentage of sequence identity.

[0017] As used herein, the term "antibody" is generally an immunoglobulin molecule consisting of two pairs of polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC). Antibody light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains can be classified as μ, δ, γ, α, or ε, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of approximately 12 or more amino acids, and the heavy chain may also include a "D" region of approximately 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain, CL. The constant domain is not directly involved in the binding of antibodies to antigens, but exhibits a variety of effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The VH and VL regions can also be subdivided into regions with high variability (called complementarity determining regions (CDRs)), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. The variable regions (VH and VL) in each heavy chain and light chain respectively form the antigen binding site.

[0018] In some embodiments, the present invention provides "anti-Norovirus type 1 VP1 protein antibodies" that are monoclonal antibodies. A monoclonal antibody refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies comprising the antibody population are identical except for possible naturally occurring mutations that may be present in minor amounts. The modifier "monoclonal" refers to the characteristic of the antibody as being obtained from a substantially homogeneous antibody population and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in accordance with the present invention can be prepared by the hybridoma (murine or human) method first described by Kohler et al., Nature, 256:495 (1975), or can be prepared by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). "Monoclonal antibodies" can also be isolated from phage antibody libraries using, for example, the techniques described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J Mol. Biol., 222:581-597 (1991). It should also be understood that certain embodiments of the present invention relate to compositions comprising more than one different monoclonal antibody that specifically binds to the VP1 protein, ie, polyclonal antibody compositions comprising a plurality of monoclonal antibodies with different epitope specificities.

[0019] The present invention provides a nucleic acid molecule encoding the above-mentioned monoclonal antibody.

[0020] Furthermore, the nucleic acid molecules include DNA, cDNA, mRNA, and recombinant nucleic acid.

[0021] Furthermore, the nucleotide sequence of the nucleic acid molecule is codon optimized.

[0022] As used herein, the term "nucleic acid" is intended to include polymeric forms of nucleotides of any length, containing deoxyribonucleotides, ribonucleotides, and / or their analogs, including DNA, RNA, and DNA / RNA hybrids, including DNA or RNA analogs such as those containing modified backbones (e.g., peptide nucleic acids (PNA) or phosphorothioates) or modified bases. Thus, nucleic acids of the present invention include DNA, cDNA, mRNA, recombinant nucleic acids, and the like.

[0023] The present invention provides a vector comprising the aforementioned nucleic acid molecule.

[0024] Furthermore, the vector includes retrovirus (including lentivirus), adenovirus, adeno-associated virus, herpes virus (such as herpes simplex virus), poxvirus, baculovirus, papillomavirus, papovavirus, lambda phage, M13 phage, and plasmid.

[0025] As used herein, the term "vector" refers to the process of linking the isolated nucleic acid molecule to a vector, and may refer to the direct or indirect linkage of a nucleic acid sequence to regulatory elements on the vector, as long as these regulatory elements are capable of regulating the translation and expression of the nucleic acid molecule. These regulatory elements may be derived directly from the vector itself or exogenously, i.e., not derived from the vector itself. In other words, the nucleic acid molecule and the regulatory elements are operably linked. Commonly used vectors include prokaryotic expression vectors, eukaryotic expression vectors, viral expression vectors, and the like.

[0026] In some embodiments, the selection of a specific vector will depend on the target cell and the conditions being treated. Introduction can be by standard techniques such as infection, transfection, transduction or transformation. Examples of gene transfer modes include, for example, naked DNA, CaPO4 precipitation, DEAE polydextrose, electroporation, protoplast fusion, liposome transfection, cell microinjection and viral vectors.

[0027] The present invention provides a cell, which comprises the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, and the aforementioned vector.

[0028] Furthermore, the cells include eukaryotic cells and prokaryotic cells.

[0029] Furthermore, the prokaryotic cells include bacterial cells, Escherichia coli, and Streptomyces.

[0030] Furthermore, the eukaryotic cells include yeast cells, mammalian cells, and insect cells.

[0031] Furthermore, the mammalian cells include humans, monkeys, mice, rats, hamsters, goats, sheep, cattle, pigs, dogs, and cats.

[0032] Furthermore, the eukaryotic cells include human cells, mouse cells, rat cells, hamster cells, goat cells, sheep cells or any other cells.

[0033] Furthermore, the mammalian cells include A549, WEHI, 3T3, 10T1 / 2, BHK, MDCK, COS1, COS7, BSC1, BSC40, BMT10, VERO, WI38, HeLa, 293 cells (which can express functional adenovirus E1), Saos, C2C12, L cells, HT1080, and HepG2.

[0034] Furthermore, the mammalian cells include fibroblasts, hepatocytes and myoblasts.

[0035] The present invention provides a conjugate comprising the aforementioned monoclonal antibody and a directly or indirectly conjugated linker thereof.

[0036] Furthermore, the linkable substance includes a detectable label, a drug, a toxin, a cytokine, a radionuclide, or an enzyme.

[0037] As used herein, the term "detectable label" refers to any portion that generates a measurable signal by a change in the optical, electrical, or other physical indicator of the molecular state to which the portion is coupled. Such physical indicators include spectral, photochemical, biochemical, immunochemical, electromagnetic, radiochemical, and chemical means, such as, but not limited to, fluorescence, chemiluminescence, and chemiluminescence. When used with respect to a labeled detection agent, a "direct label" is a detectable label that is attached to the detection agent by any means. When used with respect to a labeled detection agent, an "indirect label" is a detectable label that specifically binds to the detection agent. Therefore, indirect labels include the following: a specific binding partner of the detection agent. Biotin and avidin are examples of such portions that are employed, for example, by contacting a biotinylated antibody with a labeled avidin to produce an indirectly labeled antibody.

[0038] In some embodiments, the label can be selected from isotopic labels, isobaric labels, enzyme labels, colored labels, fluorescent labels, chromogenic labels, luminescent labels, radioactive labels, haptens, biotin, metal complexes, metals and colloidal gold, among which isotopic labels and isobaric labels are particularly preferred.

[0039] The present invention provides a pharmaceutical composition comprising the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned cell, and / or the aforementioned conjugate.

[0040] Furthermore, the pharmaceutical compositions are administered systemically, specifically via intranasal, intravenous, intraperitoneal, intramuscular, intradermal, subcutaneous, subdermal, transdermal, intracranial, mucosal, anal, vaginal, oral, buccal routes or they can be delivered by inhalation.

[0041] Furthermore, the pharmaceutical composition includes a pharmaceutically acceptable carrier or excipient.

[0042] In some embodiments, excipients include pharmaceutically acceptable solvents, dispersants, additives, plasticizers, and the like. Typically, these substances can be formulated in a non-toxic, inert, and pharmaceutically acceptable carrier medium. The formulated pharmaceutical composition can be administered by conventional routes, including but not limited to: intratumoral, intraperitoneal, intravenous, or topical administration. In other embodiments, the pharmaceutical composition is an aqueous pharmaceutical composition (e.g., an aqueous solution), or the Nanobody and salt are provided in water without a buffer, or contain an aqueous buffer or other type of solvent (e.g., an organic solvent).

[0043] The present invention provides any of the following applications, including:

[0044] 1) Use of the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned cell, and / or the aforementioned conjugate in the preparation of a product for detecting norovirus;

[0045] Furthermore, the norovirus includes norovirus type 1.

[0046] Furthermore, the norovirus includes norovirus type 1 VP1 protein.

[0047] Furthermore, the product includes a kit, a test paper, a nucleic acid membrane strip, a chip, a system, or a device.

[0048] 2) Use of the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned cell, and / or the aforementioned conjugate in the preparation of a pharmaceutical composition for preventing or treating norovirus;

[0049] Furthermore, the norovirus includes norovirus type 1.

[0050] Furthermore, the norovirus includes norovirus type 1 VP1 protein.

[0051] As used herein, the terms "treatment" or "prevention" refer to any type of intervention or treatment performed on an individual, or the administration of an active agent to a subject, with the intent to reverse, alleviate, ameliorate, inhibit, slow, or prevent the progression, occurrence, exacerbation, or recurrence of symptoms, complications, conditions, or biochemical markers associated with a disease. Prevention refers to administration to a subject not already suffering from the disease in order to prevent the disease from occurring or to minimize its effects if it does occur.

[0052] 3) Use of the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned cell, and / or the aforementioned conjugate in the preparation of a product for regulating the activity or level of Norovirus Type 1 VP1 protein.

[0053] Furthermore, the product includes a kit, a test paper, a nucleic acid membrane strip, a chip, a system, or a device.

[0054] The present invention provides any of the following methods, comprising:

[0055] 1) A method for producing the aforementioned monoclonal antibody, comprising: transforming the aforementioned nucleic acid molecule or the aforementioned vector into cells, culturing the cells, or directly culturing the aforementioned cells, and isolating and purifying the aforementioned monoclonal antibody from the cell culture fluid;

[0056] 2) A method for inhibiting the activity or level of norovirus type 1 VP1 protein in vitro for non-therapeutic purposes, the method comprising: administering the aforementioned monoclonal antibody, the aforementioned nucleic acid molecule, the aforementioned vector, and / or the aforementioned cell to cells or tissues containing norovirus type 1 in vitro;

[0057] 3) A method for detecting norovirus in a sample for non-diagnostic and non-therapeutic purposes, the method comprising contacting the sample with the aforementioned monoclonal antibody and / or the aforementioned conjugate, and detecting the formation of the complex or the change of the marker.

[0058] Advantages and beneficial effects of the present invention:

[0059] The present invention provides a monoclonal antibody against Norovirus Type 1 VP1 protein, which has strong affinity and high binding activity to Norovirus Type 1 VP1 protein and is an excellent monoclonal antibody for detecting and treating Norovirus. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 This is the electrophoresis diagram of monoclonal antibody 1B6;

[0061] Figure 2 This is the HPLC profile of monoclonal antibody 1B6;

[0062] Figure 3 This is a graph showing the binding activity of monoclonal antibody 1B6. DETAILED DESCRIPTION

[0063] Hereinafter, the present invention will be described in further detail with reference to the following examples. However, these examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention.

[0064] Example

[0065] 1. Recombinant expression of immunogens

[0066] The norovirus 1 VP1 protein sequence was synthesized and constructed into the pEM5.1 vector. The transfection plasmid was extracted and transfected into HEK293 cells, which were cultured for 7 days. The supernatant was harvested, purified by Ni column, and concentrated to obtain the recombinant norovirus 1 VP1 protein after buffer exchange. The recombinant norovirus 1 VP1 protein sequence was obtained from Uniprot and is as follows:

[0067] (SEQ ID NO: 16)

[0068] 2. Immunity

[0069] Immunization subjects: mice, SP2 / 0 fusion, screening, subcloning, specific steps are as follows: the first immunization is with Freund's complete adjuvant, 100 μg per mouse, intraperitoneal injection, total dose 0.5 ml / mouse, and the second immunization is performed 3 weeks later; the second immunization is with Freund's incomplete adjuvant, dose 50 μg / 0.5 ml / mouse, and the third immunization is performed 2 weeks later; cell fusion is prepared 10 days after the third injection.

[0070] Take feeder cells, press 10 5 / well, plate 10 on the day before fusion 5 Mouse immune spleen cells were fused with the prepared myeloma cells using the fusion agent PEG, and plated into a 96-cell culture plate that had been added with feeder cells, 100 μl / well.

[0071] Positive wells were screened by ELISA detection method, and recombinant norovirus type 1 VP1 protein was plated overnight; the plates were washed and blocked with skim milk powder at 37°C for 1 hour; the plates were washed and 100 μl of 96-well culture supernatant was added, and the cells were incubated at 37°C for 1 hour; the plates were washed and HRP-labeled goat anti-mouse secondary antibody was added, and the cells were incubated at 37°C for 30 minutes; the plates were washed and the color development solution was added, and the color development was carried out for 10 minutes. The stop solution was added and the OD450 value was read; the cell lines with high expression levels were screened for subcloning.

[0072] 3. Sequence Retrieval

[0073] Cells were harvested, RNA was extracted, reverse transcribed, primers designed, PCR was performed, transformation was performed, clones were selected, and sequencing was performed. The resulting antibody against the Norovirus 1 VP1 protein, clone 1B6, was obtained. The specific sequence is shown in Table 1.

[0074] Table 1

[0075]

[0076]

[0077] 4. Antibody Expression

[0078] Monoclonal Antibody Expression and Purification:

[0079] (1) The screened sequences were chemically synthesized and cloned into a eukaryotic expression vector.

[0080] (2) Amplify and extract the plasmid.

[0081] (3) The antibody-encoding plasmid was transiently transfected into HEK293 mammalian cells.

[0082] (4) Collect the supernatant and purify the monoclonal antibody using affinity chromatography.

[0083] HEK293 cells were transiently transfected using PEI, and the results showed that the expression level of antibody 1B6 was 249 mg / L.

[0084] 5. Physical and chemical properties

[0085] 5.1 Gel electrophoresis detection

[0086] A. Sample Preparation

[0087] Take 20 μL of sample and mix it evenly with 5 μL of 5× reducing buffer, heat it at 95°C for 5 min, and cool it down; take 20 μL of sample and mix it evenly with 5 μL of 5× non-reducing buffer.

[0088] B. Electrophoresis

[0089] Prepare gel, add appropriate amount of electrophoresis buffer, add sample, and perform electrophoresis.

[0090] C. Dyeing and bleaching

[0091] After electrophoresis, place the gel in an appropriate amount of Coomassie Brilliant Blue staining solution and stain at room temperature for 1 hour or longer. Pour off the staining solution and add an appropriate amount of Coomassie Brilliant Blue destaining solution. Destain at room temperature for 4-24 hours. After destaining, soak the gel in ddH2O and compare it to the unstained gel with a marker protein. Cut the gel containing the desired protein fraction and collect it. The protein to be purified can then be separated from the gel.

[0092] 5.2 HPLC detection of monoclonal antibody purity

[0093] A. Mobile phase preparation

[0094] Add potassium hydrogen phosphate dihydrate, potassium dihydrogen phosphate, and potassium chloride to approximately 900 mL of purified water, stir to dissolve, and dilute to 1 L. Measure the pH with a pH meter to determine if it is between 6.2 ± 0.1. Filter through a 0.22 μm filter and store at room temperature.

[0095] B. Sample Preparation

[0096] System suitability sample: Dilute the standard to 2 mg / mL with mobile phase;

[0097] Test sample: Dilute the sample to be tested to 2 mg / mL with mobile phase.

[0098] Detection was performed under conventional chromatographic conditions.

[0099] The results are as follows Figure 1 and Figure 2 As shown, the results showed that the purity of antibody 1B6 was greater than 95% by electrophoresis and liquid chromatography detection.

[0100] 6. Binding activity

[0101] Binding activity assay of monoclonal antibodies:

[0102] A. Coating: Dilute the anti-norovirus type 1 VP1 protein to 2 μg / ml with coating solution, mix well, add to a 96-well coating plate at 100 μl / well, seal the plate, and incubate at 4°C overnight.

[0103] B. Wash the plate three times with a plate washer. No liquid should remain on the plate during the last wash. Pat the liquid on the surface of the plate dry with absorbent paper.

[0104] C. Blocking: Add 5% milk powder (0.5 g milk powder dissolved in 10 mL DPBS), 300 μL / well, incubate at 37°C for 1 h, and wash the plate three times according to step B.

[0105] D. Dilute the antibody serially, 100 μL / well, react at 37°C for 1 h, and wash the plate three times as in step B.

[0106] E. Add secondary antibody: dilute with DPBS at 1:2000, add to 96-well plate, 100 μL / well, incubate at 37°C for 1 h, and wash the plate three times as in step B.

[0107] F. Color development: Add TMB, 100 μL / well, and develop color at room temperature in the dark for 10 min.

[0108] G. Stop: Add 2N H2SO4, 100 μL / well.

[0109] H. Measure OD450 with a microplate reader within 10 minutes.

[0110] Coat VP1 protein, add antibody, add secondary antibody, develop color, read, and analyze data.

[0111] The results are as follows Figure 3 As shown, the results showed that the EC50 of antibody 1B6 was 0.00955 ng / mL, with excellent performance.

Claims

1. A monoclonal antibody against VP1 protein, wherein the heavy chain variable region of the monoclonal antibody includes complementary determining regions HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NOs: 1-3, respectively, the light chain variable region includes complementary determining regions LCDR1 and LCDR3 as shown in SEQ ID NOs: 4-5, respectively, and the light chain variable region includes complementary determining region LCDR2, and the amino acid sequence of the complementary determining region LCDR2 of the light chain variable region is NAK.

2. The monoclonal antibody according to claim 1, wherein the amino acid sequences of the framework regions FR1, FR2, FR3, and FR4 of the heavy chain variable region of the monoclonal antibody are shown in SEQ ID NOs: 6-9, respectively, and the amino acid sequences of the framework regions FR1, FR2, FR3, and FR4 of the light chain variable region of the monoclonal antibody are shown in SEQ ID NOs: 10-13, respectively.

3. The monoclonal antibody according to claim 1, wherein the amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown in SEQ ID NO: 14, and the amino acid sequence of the light chain variable region of the monoclonal antibody is shown in SEQ ID NO:

15.

4. A nucleic acid molecule encoding the monoclonal antibody according to any one of claims 1 to 3. The nucleic acid molecule of claim 4 , wherein the nucleic acid molecule comprises DNA, cDNA, mRNA, or recombinant nucleic acid. A vector comprising the nucleic acid molecule according to claim 4 .

7. The vector according to claim 6, wherein the vector comprises a retrovirus, adenovirus, adeno-associated virus, herpes virus, poxvirus, baculovirus, papillomavirus, papovavirus, lambda phage, M13 phage, or plasmid.

8. A cell comprising the monoclonal antibody according to any one of claims 1 to 3, the nucleic acid molecule according to claim 4 or 5, or the vector according to claim 6 or 7.

9. The cell of claim 8, comprising a eukaryotic cell and a prokaryotic cell.

10. The cell of claim 9, wherein the prokaryotic cell comprises a bacterial cell. The cell according to claim 10 , wherein the bacterial cell comprises Escherichia coli or Streptomyces.

12. The cell according to claim 9, wherein the eukaryotic cell comprises a yeast cell, a mammalian cell, or an insect cell.

13. The cell according to claim 12, wherein the mammalian cell is derived from humans, monkeys, mice, rats, hamsters, goats, sheep, cattle, pigs, dogs or cats.

14. The cell of claim 9, wherein the eukaryotic cell comprises a human cell, a mouse cell, a rat cell, a hamster cell, a goat cell, a sheep cell, or any other cell.

15. The cell of claim 12, wherein the mammalian cell comprises A549, WEHI, 3T3, 10T1 / 2, BHK, MDCK, COS1, COS7, BSC1, BSC40, BMT10, VERO, WI38, HeLa, 293 cells, Saos, C2C12, L cells, HT1080, HepG2.

16. The cell of claim 12, wherein the mammalian cell comprises a fibroblast, a hepatocyte, and a myoblast.

17. A conjugate comprising the monoclonal antibody according to any one of claims 1 to 3 and a directly or indirectly conjugated linker thereof; the linker comprising a detectable label or a radionuclide.

18. Use of the monoclonal antibody according to any one of claims 1 to 3, the nucleic acid molecule according to claim 4 or 5, the vector according to claim 6 or 7, the cell according to any one of claims 8 to 16, and / or the conjugate according to claim 17 in the preparation of a product for detecting norovirus.

19. The use according to claim 18, wherein the norovirus comprises norovirus type 1.

20. The use according to claim 18, wherein the norovirus comprises norovirus type 1 VP1 protein.

21. A method for producing the monoclonal antibody according to any one of claims 1 to 3, comprising: The nucleic acid molecule according to claim 4 or 5 or the vector according to claim 6 or 7 is transformed into cells, and the cells are cultured, or the cells according to any one of claims 8 to 16 are directly used for culture, and the monoclonal antibody according to any one of claims 1 to 3 is isolated and purified from the cell culture fluid.

22. A method for detecting norovirus in a sample for non-diagnostic and non-therapeutic purposes, the method comprising contacting the sample with the monoclonal antibody according to any one of claims 1 to 3, and / or the conjugate according to claim 17, and detecting the formation of a complex or the change of a marker.

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