CN111592595A - Neutralizing antibody against novel coronavirus SARS-Cov-2 and application thereof - Google Patents

Abstract

The present invention relates to a neutralizing antibody against novel coronavirus SARS-Cov-2 and its application. The antibody has at least one of heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, and light chain CDR3. The antibody can be used to prepare diagnostic reagents or diagnostic kits, medicines or pharmaceutical compositions for detecting, preventing and treating COVID-19. The present invention uses phage display technology to target SARS-Cov-2-RBD and SARS-Cov-1-RBD for differential antibody screening, and obtains neutralizing antibodies against the new coronavirus SARS-Cov-2, which can block SARS The combination of ‑Cov‑2‑RBD and ACE2 positive cells has a significant virus neutralization effect on the SARS‑Cov‑2 pseudovirus, providing an effective alternative antibody drug for the prevention and treatment of COVID‑19.

Description

Neutralizing antibody against novel coronavirus SARS-Cov-2 and its application

technical field

The invention relates to a neutralizing antibody against novel coronavirus SARS-Cov-2 and its application, belonging to the technical field of biomedicine.

Background technique

The 2019 novel coronavirus pneumonia (COVID-19) is caused by the infection of the new coronavirus (Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2). Currently, more than 2 million people have been infected worldwide, and the fatality rate exceeds 6% ( https:// covid19.who.int/ ), is a major infectious disease that endangers human health. SARS-CoV-2 is a positive-strand single-stranded RNA virus, belonging to beta coronavirus ^[1] , and its nucleic acid homology with SARS-Cov-1 is 79.5% ^[2] , and its natural host is unknown. Both SARS-CoV-2 and SARS-Cov-1 infect host cells by binding to angiotensin-converting enzyme 2 (ACE2) on the host cell membrane ^[3] .

ACE2 is an angiotensin-converting enzyme, and its substrates are type I and type II angiotensin. Under physiological conditions, ACE2 resists the vasoconstrictive effect caused by ACE1 by hydrolyzing the substrate. SARS-Cov-2 binds to ACE2 through the receptor binding domain (RBD) of Spike glycoprotein S1 subunit on the virus coat, and then invades host cells ^[4] .

The protein structure of the complex of SARS-Cov-2-RBD and ACE2 has been analyzed. Compared with SARS-Cov-1, both RBDs bind to ACE2 at the same angle, through 17 amino acid residues and 20 of ACE2. Amino acid residues form interactions ^[3,5] . Subtle differences in the contact interface of the two RBDs with ACE2 lead to differences in the affinity of SARS-Cov-1 and SARS-Cov-2 for ACE2, which may be beneficial for follow-up studies.

Currently, there are no SARS-CoV-2 virus-specific vaccines and neutralizing antibodies for clinical treatment. Therefore, obtaining neutralizing human monoclonal antibodies through screening is an urgent need for the prevention and treatment of COVID-19.

SUMMARY OF THE INVENTION

The main purpose of the present invention is to overcome the problems existing in the prior art, and to provide a neutralizing antibody against the novel coronavirus SARS-Cov-2, which has high-efficiency antiviral ability against the novel coronavirus SARS-Cov-2. At the same time, the application of the antibody is also provided.

The technical scheme that the present invention solves its technical problem is as follows:

A neutralizing antibody against the novel coronavirus SARS-Cov-2, the antibody comprises a heavy chain and a light chain, wherein the antibody has at least one of the following technical characteristics:

i, the heavy chain includes the heavy chain CDR1, and its amino acid sequence is: GFTFSSYA;

ii, the heavy chain includes the heavy chain CDR2, and its amino acid sequence is: SIASSGYYTD;

iii, the heavy chain includes heavy chain CDR3, and its amino acid sequence is: KDADS;

iv, the light chain includes light chain CDR1, and its amino acid sequence is: ISSYL;

v, the light chain includes the light chain CDR2, and its amino acid sequence is: AASYL;

vi. The light chain includes light chain CDR3, and its amino acid sequence is: AYSAPS.

Preferably, the antibody has at least one of the following technical characteristics:

i. The heavy chain includes heavy chain CDR1, whose amino acid sequence is: GFTFSSYA; the heavy chain includes heavy chain CDR2, whose amino acid sequence is: SIASSGYYTD; the heavy chain also includes heavy chain CDR3, whose amino acid sequence is: KDADS ;

ii, the light chain includes light chain CDR1, and its amino acid sequence is: ISSYL; the light chain includes light chain CDR2, and its amino acid sequence is: AASYL; the light chain also includes light chain CDR3, and its amino acid sequence is: AYSAPS .

Preferably, the amino acid sequence of the antibody is shown in SEQ ID NO:2.

Preferably, the antibody is a VH single-domain structure antibody, Fab fragment, Fab' fragment, F(ab)' ₂ fragment, single-chain variable fragment scFv, disulfide stabilized variable fragment dsFv, IgG molecule, or bispecific antibodies.

Preferably, the antibody has a label including fluorescent labels, enzymatic labels, and radioactive labels.

The present invention also provides:

Nucleic acid encoding the neutralizing antibody against the novel coronavirus SARS-Cov-2 described above.

Preferably, the sequence of the nucleic acid is shown in SEQ ID NO:1.

The present invention also provides:

Use of the aforementioned neutralizing antibody against novel coronavirus SARS-Cov-2 for preparing diagnostic reagents or diagnostic kits, medicines or pharmaceutical compositions.

Use of the aforementioned nucleic acid for preparing neutralizing antibodies, drugs or pharmaceutical compositions against novel coronavirus SARS-Cov-2.

Wherein, the medicine or pharmaceutical composition has a neutralizing antiviral effect against the novel coronavirus SARS-Cov-2.

The present invention utilizes phage display technology to screen differential antibodies by targeting SARS-Cov-2-RBD and SARS-Cov-1-RBD to obtain neutralizing antibodies against the new coronavirus SARS-Cov-2. It can block the combination of SARS-Cov-2-RBD and ACE2 positive cells, has a significant virus neutralization effect on SARS-Cov-2 pseudovirus, and provides an effective alternative antibody drug for the prevention and treatment of COVID-19, Has potential clinical application prospects.

Description of drawings

Fig. 1 is a graph showing the binding of the enriched phage to the antigenic protein by ELISA in Example 1 of the present invention.

Fig. 2 is the specific binding detection (ELISA) diagram of phage to SARS-Cov-2-RBD protein in Example 2 of the present invention.

FIG. 3 and FIG. 4 are schematic diagrams of the expression vector of Example 3 of the present invention, respectively.

FIG. 5 is a graph showing the results of SDS-PAGE in Example 3 of the present invention.

FIG. 6 is a graph showing the results of affinity analysis in Example 4 of the present invention.

FIG. 7 is an analysis diagram of the antibody blocking effect of Example 5 of the present invention.

FIG. 8 is a graph showing the evaluation of the anti-pseudovirus neutralization effect of the antibody of Example 6 of the present invention.

Detailed ways

The present invention will be described in further detail below in conjunction with the embodiments. However, the invention is not limited to the examples given. The methods used are conventional methods unless otherwise specified, and the reagents and materials used are commercially available unless otherwise specified.

Example 1. Screening of fully human antibodies targeting SARS-Cov-2-RBD

Using phage display technology, the SARS-Cov-2-RBD-his protein was used as the positive antigen, and the SARS-Cov-1-RBD-hFc was used as the negative antigen. Differential screening was performed in 1.47x10 ⁸ ).

Immunoplates were coated with 50 μg/ml SARS-Cov-2-RBD his antigen and SARS-Cov-1-RBD hFc antigen at 4°C overnight; blocked with PBS solution containing 5% nonfat dry milk and 0.1% Tween-20 at room temperature Immune plate for 1 hour; phage library was mixed 1:1 with ¹⁰ % skimmed milk powder in PBS, incubated at room temperature for 2 hours, added to the blocked SARS-Cov-1-RBD hFc antigen immune plate (100 μl/well), Incubate at room temperature for 1 hour to perform pre-adsorption of negative antigens; after pre-adsorption, the supernatant is transferred to the blocked SARS-Cov-2-RBD his antigen immune plate (100 μl/well), and incubated at room temperature for 1 hour. The immune plate was washed 20 times with 0.1% Tween-20 in PBS; 100 μl of 100 mM Triethylamine was eluted at room temperature for 30 minutes; the eluted phages were infected with TG1 cells in logarithmic growth phase, amplified and recovered for the next round of panning. The enrichment of positive phages was analyzed by ELISA after panning.

The specific process of ELISA detection is as follows: with 5 μg/ml of SARS-Cov-2-RBD his antigen and negative control protein GPC5 his respectively at 4 ° C to coat the immune plate overnight; with 3% skimmed milk powder, 0.1% Tween-20 The immune plate was blocked with PBS solution for 1 hour at room temperature; the enriched phages in each round were amplified and recovered, incubated at room temperature with 6% nonfat dry milk PBS at a ratio of 1:1 for 2 hours, and added to the blocked immune plate (100 μl/well) , incubate for 1 hour at room temperature; wash the immunoplate 5 times with 0.1% Tween-20 in PBS; mix HRP/Anti-M13 Monoclonal conjugate with 5% nonfat dry milk, 0.05% Tween-20 in PBS at a ratio of 1:4000 , add it to the washed immune plate (50 μl/well), incubate at room temperature for 1 hour; wash the immune plate 5 times with 0.05% Tween-20 in PBS; add TMB chromogenic solution to the immune plate (100 μl/well) at room temperature After 3 minutes of color development, 0.5M sulfuric acid was added to stop the color development (100 μl/well); the absorbance was detected with an enzyme-linked immunosorbent assay at a wavelength of 450 nm, and the affinity of the phage after each round of amplification was analyzed.

The results are shown in Figure 1. After four rounds of enrichment, the affinity of the enriched phage population for the SARS-Cov-2-RBD-his antigen was significantly increased.

Single clones were randomly picked from the fourth round of enriched phage population and tested for their binding properties to SARS-Cov-2-RBD-his. It was found that one antibody sequence (4A3) was significantly enriched.

After sequencing, the DNA sequence of the antibody is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2.

SEQ ID NO: 1:

gaggtgcagctgttggagtctgggggaggcttggtacagcctggggggtccctgagactctcctgt gcagcctctggattcacctttagcagctatgccatgagctgggtccgccaggctccagggaagggg ctggagtgggtctcatctattgcttcttctggttattatacagattacgcagactccgtgaagggc cggttcaccatctccagagacaattccaagaacacgctgtatctgcaaatgaacagcctgagagcc gaggacacggccgtatattactgtgcgaaagatgctgattcttttgactactggggccagggaacc ctggtcaccgtctcgagcggtggaggcggttcaggcggaggtggcagcggcggtggcgggtcggac atccagatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgc cgggcaagtcagagcattagcagctatttaaattggtatcagcagaaaccagggaaagcccctaag ctcctgatctatgctgcatcctatttgcaaagtggggtcccatcaaggttcagtggcagtggatct gggacagatttcactctcaccatcagcagtctgcaacctgaagattttgcaacttactactgtcaa caggcttattctgctccttctacgttcggccaagggaccaaggtggaaatcaaa。

SEQ ID NO: 2:

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSIASSGYYTDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDADSFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASYLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAYSAPSTFGQGTK.

In this amino acid sequence, the included CDR regions are as follows: amino acid residues 26-33 (ie GFTFSSYA) are heavy chain CDR1, amino acid residues 50-59 (ie SIASSGYYTD) are heavy chain CDR2, amino acid residues 98-102 (ie KDADS) ) is heavy chain CDR3; amino acid residues 160-164 (ie ISSYL) are light chain CDR1, amino acid residues 181-185 (ie AASYL) are light chain CDR2, amino acid residues 222-227 (ie AYSAPS) are light chain CDR3 .

In addition, the antibody format can be selected from VH single domain structure antibodies, Fab fragments, Fab' fragments, F(ab)' ₂ fragments, single chain variable region fragments (scFv), disulfide stabilized variable region fragments (dsFv) ), IgG molecules, or bispecific antibodies. The antibody can optionally be labeled, including fluorescent labels, enzymatic labels, and radioactive labels.

Example 2. Antigen specificity analysis of antibodies

In this example, ELISA was used to detect the binding of the 4A3 phage of Example 1 to the SARS-Cov-2-RBD protein.

The specific process is as follows: use 5 μg/ml SARS-Cov-1-RBD-hFc and SARS-Cov-2-RBD-hFc to coat the immune plate overnight at 4°C; The PBS solution of 0.05% Tween-20 was used to block the immune plate for 1 hour at room temperature; the phage of 4A3 was added to the blocked immune plate (50 μl/well), and incubated at room temperature for 1 hour; the immune plate was washed 3 times with 0.05% Tween-20 in PBS (340 μl/well). ); Mix HRP/Anti-M13 Monoclonal conjugate with PBS solution containing 5% nonfat milk powder and 0.05% Tween-20 at a ratio of 1:4000, add it to the washed immune plate (50μl/well), and incubate at room temperature for 1 hour; Wash the immune plate 5 times with 0.05% Tween-20 in PBS; add TMB color development solution to the immune plate (100 μl/well), and after 3 minutes of color development at room temperature, add 0.5M sulfuric acid to stop the color development (100 μl/well); Absorbance values were detected at 450 nm wavelength with an enzyme-linked immunosorbent assay.

As shown in Figure 2, the results showed that the 4A3 antibody specifically recognized the SARS-Cov-2-RBD-hFc protein, but not the SARS-Cov-1-RBD-hFc protein.

Example 3. Expression and purification of antibodies

The heavy chain variable region sequence and light chain variable region sequence of the 4A3 scFv sequence were inserted into the pFUSE-CHIg-HG1 and pFUSE2-CLIg-hk vectors (Invivogen, San Diego, CA), respectively, to construct an adult IgG1 expression plasmid, as shown in Figure 3 , as shown in Figure 4. The above plasmids were co-transfected in 293T cells, and the supernatant was collected and purified by protein A-Agarose separation column. The purity of the antibody was detected by SDS-PAGE, as shown in Figure 5.

The specific process is as follows: insert the heavy chain variable region sequence and light chain variable region sequence of the 4A3 scFv sequence into pFUSE-CHIg-HG1 and pFUSE2-CLIg-hk (Invivogen, San Diego, CA), respectively, to construct an adult IgG expression plasmid. 5 million HEK293T cells were seeded in a cell culture dish with DMEM medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 0.1 mg/ml streptomycin, and cultured in a 5% CO2, 37°C incubator. When the cell density reached 60-80%, 5 μg pFUSE-4A3 VH and 5 μg pFUSE-4A3 VL plasmids were co-transfected into HEK293T cells using PEI; the supernatant was collected.

The collected supernatant was centrifuged at 3500 rpm and 4 °C for 20 minutes, and filtered with a 0.45 μm microporous filter to further remove debris; and column separation and purification of 4A3 IgG recombinant protein. The protein concentration was determined by BCA method, and 3 μg of 4A3 IgG recombinant protein was subjected to polyacrylamide gel electrophoresis to obtain a band of 4A3 IgG recombinant protein.

Example 4. Antibody affinity analysis

Using SPR experiment, the affinity of 4A3 and SARS-Cov-2-RBD his protein was determined.

4A3 antibody affinity analysis was performed by Nanjing GenScript, and surface plasmon resonance analysis (SPR) was performed with Biacore T200, GR18010468 (GE Healthcare). Specific steps are as follows:

First, the antibody 4A3 was immobilized on the Series S Sensor Chip Protein A chip (GE Healthcare), and then the SARS-Cov-2-RBD protein with a concentration gradient of 1.25nM-40nM was injected into the chip, and the analysis was performed at a constant temperature of 25°C using The buffer was HBS-EP+: 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% P20, pH 7.4 (Lot. No. 30393) (GE Healthcare); flow rate was 10 μl/min. The binding curves are shown in Figure 6. The antibody concentrations of the five curves in the figure from top to bottom are 40, 20, 10, 5, 2.5, and 1.25 nM, respectively, and the curves of different concentrations constitute the kinetic curves shown in the figure. Calculation of binding kinetic constants was performed using Biacore T200 Evaluation software version 3.1 software. The results showed that the affinity test results of 4A3 antibody and SARS-Cov-2-RBD his protein were ka=1.91E+06, kd=1.90E-03, and KD was 3.62nM.

Example 5. Blockade of the binding of antibodies to SARS-Cov-2-RBD and ACE2-CHO cells

After pre-incubating 4A3IgG, control IgG, M396 antibody (this is SARS-Cov-1 neutralizing antibody) with SARS-Cov-2-RBD-hFc protein for 1 hour at room temperature, 10 ⁶ ACE2-CHO cells were added to Mix the system and incubate on ice for 1 hour. Cells were washed with PBS, added with Goat anti-human PE at 1:200, and incubated on ice for 1 hour. The cells were washed with PBS, and the BD FACSCalibur was used to detect the binding of SARS-Cov-2-RBD-hFc to ACE2-CHO cells.

The results are shown in Figure 7. After incubation with SARS-Cov-2-RBD protein, 4A3 antibody can significantly inhibit the binding of SARS-Cov-2-RBD to ACE2-CHO cells, while SARS-Cov-1-RBD neutralizes the Antibody M396 cannot block the binding of SARS-Cov-2-RBD to ACE2-CHO cells

Example 6. Virus neutralization experiment

The VSV-G protein gene was replaced with the SARS-Cov-2 spike gene in the lentiviral packaging system, and 293T cells were co-transfected with the pLVX-EGFP-Luciferase reporter gene (ie, pseudovirus transfection), and the virus supernatant was collected for 48 hours. , 1:1 diluted for use. ACE2-CHO cells were seeded at 10 ⁴ /well in a 96-well plate and cultured overnight. After pre-incubating the serially diluted antibody and virus supernatant at 37°C for 1 hour, it was added to the ACE2-CHO cell culture plate. After 48 hours, Luciferase activity was detected.

As shown in Figure 8, the results showed that the 4A3 antibody could significantly inhibit the infection of ACE2-CHO cells by pseudovirus with an IC ₅₀ of 0.28 μg/ml.

Each antibody of the present invention can specifically bind/recognize SARS-Cov-2-RBD his antigen, has good affinity for SARS-Cov-2-RBD his protein, can effectively inhibit SARS-Cov-2 invading cells, and has the ability to become Important application value of COVID-19 prevention and treatment drugs.

In addition to the above-described embodiments, the present invention may also have other embodiments. All technical solutions formed by equivalent replacement or equivalent transformation fall within the protection scope of the present invention.

references

[1]. Zhu, N., et al., A Novel Coronavirus from Patients with Pneumonia in China, 2019. N Engl J Med, 2020.382(8):727-733.

[2]. Zhou, P., et al., A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature, 2020.579(7798):270-273.

[3]. Lan, J., et al., Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature, 2020.

[4]. Hoffmann, M., et al., SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell, 2020.181(2):271-280e278.

[5]. Li, F., et al., Structure of SARS coronavirus spike receptor-bindingdomain complexed with receptor. Science, 2005. 309(5742): 1864-1868.

sequence listing

<110> Nanjing Medical University

<120> Neutralizing antibody against novel coronavirus SARS-Cov-2 and its application

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 714

<212> DNA

<213> Artificial Sequence

<400> 1

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccaggggaagg ggctggagtg ggtctcatct attgcttctt ctggttatta tacagattac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatgct 300

gattcttttg actactgggg ccagggaacc ctggtcaccg tctcgagcgg tggaggcggt 360

tcaggcggag gtggcagcgg cggtggcggg tcggacatcc agatgaccca gtctccatcc 420

tccctgtctg catctgtagg agacagagtc accatcactt gccgggcaag tcagagcatt 480

agcagctatt taaattggta tcagcagaaa ccaggaaag cccctaagct cctgatctat 540

gctgcatcct atttgcaaag tggggtccca tcaaggttca gtggcagtgg atctgggaca 600

gatttcactc tcaccatcag cagtctgcaa cctgaagatt ttgcaactta ctactgtcaa 660

caggcttatt ctgctccttc tacgttcggc caagggacca aggtggaaat caaa 714

<210> 2

<211> 238

<212> PRT

<213> Artificial Sequence

<400> 2

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ala Ser Ser Gly Tyr Tyr Thr Asp Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asp Ala Asp Ser Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

115 120 125

Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala

130 135 140

Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile

145 150 155 160

Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys

165 170 175

Leu Leu Ile Tyr Ala Ala Ser Tyr Leu Gln Ser Gly Val Pro Ser Arg

180 185 190

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

195 200 205

Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Tyr Ser

210 215 220

Ala Pro Ser Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

225 230 235

Claims (10)

1. a kind of neutralizing antibody against novel coronavirus SARS-Cov-2, described antibody comprises heavy chain and light chain, it is characterized in that, described antibody has at least one of following technical characteristics: i, the heavy chain includes the heavy chain CDR1, and its amino acid sequence is: GFTFSSYA; ii, the heavy chain includes the heavy chain CDR2, and its amino acid sequence is: SIASSGYYTD; iii, the heavy chain includes heavy chain CDR3, and its amino acid sequence is: KDADS; iv, the light chain includes light chain CDR1, and its amino acid sequence is: ISSYL; v, the light chain includes the light chain CDR2, and its amino acid sequence is: AASYL; vi. The light chain includes light chain CDR3, and its amino acid sequence is: AYSAPS. 2. The neutralizing antibody according to claim 1, wherein the antibody has at least one of the following technical characteristics: i. The heavy chain includes heavy chain CDR1, whose amino acid sequence is: GFTFSSYA; the heavy chain includes heavy chain CDR2, whose amino acid sequence is: SIASSGYYTD; the heavy chain also includes heavy chain CDR3, whose amino acid sequence is: KDADS ; ii, the light chain includes light chain CDR1, and its amino acid sequence is: ISSYL; the light chain includes light chain CDR2, and its amino acid sequence is: AASYL; the light chain also includes light chain CDR3, and its amino acid sequence is: AYSAPS . 3. The neutralizing antibody according to claim 1, wherein the amino acid sequence of the antibody is shown in SEQ ID NO:2. 4. The neutralizing antibody according to claim 1, wherein the antibody is a VH single-domain structure antibody, Fab fragment, Fab' fragment, F(ab)' ₂ fragment, single-chain variable region fragment scFv , a disulfide stabilized variable region fragment dsFv, an IgG molecule, or a bispecific antibody. 5 . The neutralizing antibody according to claim 1 , wherein the antibody has a label, and the label includes a fluorescent label, an enzymatic label, and a radioactive label. 6 . 6. The nucleic acid encoding the neutralizing antibody of the anti-novel coronavirus SARS-Cov-2 according to any one of claims 1 to 5. 7. The nucleic acid according to claim 6, wherein the sequence of the nucleic acid is shown in SEQ ID NO:1. 8. Use of the neutralizing antibody against the novel coronavirus SARS-Cov-2 described in any one of claims 1 to 5 for the preparation of diagnostic reagents or diagnostic kits, medicines or pharmaceutical compositions. 9. the purposes of the nucleic acid described in claim 6 or 7 for preparing the neutralizing antibody, medicine or pharmaceutical composition of anti-novel coronavirus SARS-Cov-2. 10. The use according to claim 8 or 9, wherein the medicine or the pharmaceutical composition has a neutralizing antiviral effect against the novel coronavirus SARS-Cov-2.

Images