Human-derived neutralizing antibody or antigen-binding fragment thereof and use thereof

Abstract

The application is a divisional application of application No. 202210313522.1, and relates to a human neutralizing antibody or an antigen binding fragment thereof and application thereof, which comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 with the amino acid sequences shown in SEQ ID NO: 21, 22 and 23 respectively; and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 with the amino acid sequences shown in SEQ ID NO: 24, 25 and 26 respectively. The neutralizing antibody YB13-208 of the application can effectively inhibit SARS-CoV-2 pseudovirus infection, and also has good neutralizing activity on SARS-CoV-2 true virus.

Description

[0001] This invention is a divisional application of Chinese application filed on March 28, 2022, with application number 202210313522.1 and entitled "A human neutralizing antibody or its antigen-binding fragment and its application". Technical Field

[0002] This invention relates to the field of biomedicine, specifically to a human neutralizing antibody or its antigen-binding fragment and its application, and more specifically, to a human neutralizing antibody or its antigen-binding fragment that binds to the coronavirus S protein, a polypeptide, a polynucleotide, a nucleic acid construct, an expression vector, a transformed cell, a pharmaceutical composition, and their application in the preparation of drugs for the prevention, treatment, detection, or diagnosis of COVID-19 infection. Background Technology

[0003] Coronavirus disease 2019 (COVID-19), caused by a novel coronavirus called Severe Acute Respiratory Syndrome (SARS-CoV-2), has spread severely worldwide. Currently, COVID-19 has affected more than 100 countries and regions globally, infecting over 200 million people and directly causing more than 4.4 million deaths. SARS-CoV-2 is an RNA virus with a high mutation rate in its surface protein amino acids. Multiple variant strains have been reported, and with the proliferation of these variants, many countries worldwide are experiencing a resurgence of the epidemic, showing a surge in cases, with hundreds of thousands of new confirmed infections daily globally. The recently emerged variant, Omicron, has more than 30 mutations in its spike (S) glycoprotein, including as many as 15 mutations in the receptor-binding domain (RBD), posing a serious threat to the effectiveness of current vaccines and monoclonal antibody drugs. The spread and continued impact of the epidemic threatens human life and health, severely affecting normal socio-economic activities.

[0004] The S protein of SARS-CoV-2 mediates viral membrane fusion and receptor recognition. Specifically, the S1 subunit of the spike (S) glycoprotein, located at the N-terminus, is responsible for viral attachment, while its RBD directly binds to the ACE2 receptor on host cells. The S2 subunit (C-terminus) is responsible for membrane fusion. Currently, most SARS-CoV-2 vaccines under development target the spike (S) glycoprotein and have the ability to induce high levels of neutralizing antibodies (NAbs) targeting either the S protein or its RBD. However, considering the long duration of clinical trials and the uncertainty of vaccine efficacy in humans, researching SARS-CoV-2 neutralizing antibodies with expected efficacy and safety is also crucial for COVID-19 treatment strategies. Several SARS-CoV-2-specific neutralizing antibody drugs have been approved or are currently in clinical trials.

[0005] Most existing neutralizing antibodies against SARS-CoV are traditional monoclonal antibodies isolated from recovered patients. However, the broad-spectrum neutralizing activity of these antibodies is often limited by factors such as viral strain mutations. For example, Eli Lilly's Bamlanivimab (LY-CoV555) (Jones et al., 2020) and Junshi Biosciences' Etesevimab (JS016 or LY-CoV016) (Shi et al., 2020), neutralizing antibodies isolated from convalescent plasma of SARS-CoV-2 patients, were granted emergency use authorization by the FDA in February 2021. However, due to their insignificant efficacy against the Brazilian Gamma and South African Beta variants, their supply was suspended on June 25, 2021. Representative antibodies CR3022 and S309 are cross-reactive antibodies against SARS-CoV and SARS-CoV-2. However, the neutralizing activity of S309 has been found to be impaired against variants, and whether the efficacy of CR3022 is also affected remains to be determined. Antibody cocktail therapy is a combination of two clonal antibody drugs, casirivimab and imdevimab. It is unclear whether it can combat the mutations of COVID-19, and its safety and efficacy need to be evaluated.

[0006] However, SARS-CoV-2 is an RNA virus with a high mutation rate in its surface proteins, and several variant strains have been reported, such as Delta, Lambda, and Kappa. Recently, a new variant strain, Omicron, has emerged. These variant strains are mostly more infectious, have stronger replication capabilities, higher viral loads, and can evade the neutralizing ability of neutralizing antibodies. Currently, antibody drugs show a significant decrease in neutralizing antibody efficacy against the recently emerged Omicron (B.1.1.529) variant strain. Therefore, screening for a broad-spectrum neutralizing antibody that can prevent infection by known and future variants is urgently needed. Summary of the Invention

[0007] Purpose of the invention

[0008] The purpose of this invention is to provide a human neutralizing antibody or its antigen-binding fragment and its applications. More specifically, it relates to a human neutralizing antibody or its antigen-binding fragment that binds to the coronavirus S protein, a polypeptide, a polynucleotide, a nucleic acid construct, an expression vector, transformed cells, a pharmaceutical composition, and their applications in the preparation of drugs for the prevention, treatment, detection, or diagnosis of SARS-CoV-2 infection. The human neutralizing antibody of this invention can effectively neutralize various SARS-CoV-2 variants, including the original SARS-CoV-2 strain (WT), the variant Beta (B.1.351), the variant Delta (B.1.617.2), and the variant Omicron (B.1.1.529), meeting my country's pharmaceutical needs and providing an important tool for scientifically responding to the complex and ever-changing epidemic.

[0009] Solution

[0010] To achieve the above objectives, the present invention provides the following technical solution:

[0011] In a first aspect, the present invention provides a human neutralizing antibody or antigen-binding fragment thereof that binds to the coronavirus S protein, comprising a heavy chain variable region and / or a light chain variable region.

[0012] The heavy chain variable region includes:

[0013] (I) HCDR1, HCDR2, and HCDR3 with amino acid sequences as shown in SEQ ID NO:1 (i.e., SNYMH), SEQ ID NO:2 (i.e., VLYAGGSAFYADSVKG), and SEQ ID NO:3 (i.e., CARGLGDYLDSW), respectively; or HCDR1, HCDR2, and HCDR3 having 1, 2, or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:1 (i.e., SNYMH), SEQ ID NO:2 (i.e., VLYAGGSAFYADSVKG), and SEQ ID NO:3 (i.e., CARGLGDYLDSW), respectively; or

[0014] (II) HCDR1, HCDR2, and HCDR3 with amino acid sequences as shown in SEQ ID NO:11 (i.e., GFSFITY), SEQ ID NO:12 (i.e., SSNILS), and SEQ ID NO:13 (i.e., CARTRSRSVRNCTSATCPVDAFDLW), respectively; or HCDR1, HCDR2, and HCDR3 having 1, 2, or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:11 (i.e., GFSFITY), SEQ ID NO:12 (i.e., SSNILS), and SEQ ID NO:13 (i.e., CARTRSRSVRNCTSATCPVDAFDLW), respectively; or

[0015] (III) HCDR1, HCDR2 and HCDR3 with amino acid sequences as shown in SEQ ID NO:21 (i.e., TSGVSVG), SEQ ID NO:22 (i.e., LIYWDDDKRYSPSLTS) and SEQ ID NO:23 (i.e., PRYYGDSSGYYWI); or HCDR1, HCDR2 and HCDR3 with 1, 2 or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:21 (i.e., TSGVSVG), SEQ ID NO:22 (i.e., LIYWDDDKRYSPSLTS) and SEQ ID NO:23 (i.e., PRYYGDSSGYYWI);

[0016] The light chain variable region includes:

[0017] (I) LCDR1, LCDR2, and LCDR3 with amino acid sequences as shown in SEQ ID NO:4 (i.e., RASQGIGSWLA), SEQ ID NO:5 (i.e., AASTLQS), and SEQ ID NO:6 (i.e., QQANSVLALT), respectively; or LCDR1, LCDR2, and LCDR3 having 1, 2, or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:4 (i.e., RASQGIGSWLA), SEQ ID NO:5 (i.e., AASTLQS), and SEQ ID NO:6 (i.e., QQANSVLALT), respectively; or

[0018] (II) LCDR1, LCDR2, and LCDR3 with amino acid sequences as shown in SEQ ID NO:14 (i.e., RSSQSLLRSNGYNYLD), SEQ ID NO:15 (i.e., LGSNRAS), and SEQ ID NO:16 (i.e., MQALQTPYT), respectively; or LCDR1, LCDR2, and LCDR3 having 1, 2, or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:14 (i.e., RSSQSLLRSNGYNYLD), SEQ ID NO:15 (i.e., LGSNRAS), and SEQ ID NO:16 (i.e., MQALQTPYT), respectively; or

[0019] (III) HCDR1, HCDR2 and HCDR3 with amino acid sequences as shown in SEQ ID NO:24 (i.e., SGDALPKQYAY), SEQ ID NO:25 (i.e., KTSERPS) and SEQ ID NO:26 (i.e., QSADSSGFYV); or HCDR1, HCDR2 and HCDR3 with 1, 2 or 3 amino acid differences from the amino acid sequences shown in SEQ ID NO:24 (i.e., SGDALPKQYAY), SEQ ID NO:25 (i.e., KTSERPS) and SEQ ID NO:26 (i.e., QSADSSGFYV).

[0020] The coronavirus of this invention mainly refers to SARS-CoV-2, but it may also have neutralizing activity against other coronaviruses. The S protein in this invention mainly refers to the S1 antigen of SARS-CoV-2.

[0021] In some embodiments, the human neutralizing antibody or its antigen-binding fragment comprises a heavy chain variable region and / or a light chain variable region.

[0022] The heavy chain variable region includes:

[0023] (I) The amino acid sequences are HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:1 (i.e., SNYMH), SEQ ID NO:2 (i.e., VLYAGGSAFYADSVKG), and SEQ ID NO:3 (i.e., CARGLGDYLDSW), respectively; or

[0024] (II) The amino acid sequences are HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO:11 (i.e., GFSFITY), SEQ ID NO:12 (i.e., SSNILS), and SEQ ID NO:13 (i.e., CARTRSRSVRNCTSATCPVDAFDLW), respectively; or

[0025] (III) The amino acid sequences are HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:21 (i.e., TSGVSVG), SEQ ID NO:22 (i.e., LIYWDDDKRYSPSLTS) and SEQ ID NO:23 (i.e., PRYYGDSSGYYWI), respectively.

[0026] The light chain variable region includes:

[0027] (I) The amino acid sequences are as shown in SEQ ID NO:4 (i.e., RASQGIGSWLA), SEQ ID NO:5 (i.e., AASTLQS), and SEQ ID NO:6 (i.e., QQANSVLALT), respectively, for LCDR1, LCDR2, and LCDR3; or

[0028] (II) The amino acid sequences are as shown in SEQ ID NO:14 (i.e., RSSQSLLRSNGYNYLD), SEQ ID NO:15 (i.e., LGSNRAS), and SEQ ID NO:16 (i.e., MQALQTPYT), respectively, for LCDR1, LCDR2, and LCDR3; or

[0029] (III) The amino acid sequences are HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:24 (i.e., SGDALPKQYAY), SEQ ID NO:25 (i.e., KTSERPS) and SEQ ID NO:26 (i.e., QSADSSGFYV), respectively.

[0030] In some embodiments, the human neutralizing antibody or its antigen-binding fragment comprises:

[0031] (I) Heavy chain variable region, the amino acid sequence of which has HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:1 (i.e., SNYMH), SEQ ID NO:2 (i.e., VLYAGGSAFYADSVKG) and SEQ ID NO:3 (i.e., CARGLGDYLDSW), respectively; and light chain variable region, the amino acid sequence of which has LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:4 (i.e., RASQGIGSWLA), SEQ ID NO:5 (i.e., AASTLQS) and SEQ ID NO:6 (i.e., QQANSVLALT), respectively; or

[0032] (II) Heavy chain variable region, the amino acid sequence of which has HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:11 (i.e., GFSFITY), SEQ ID NO:12 (i.e., SSNILS) and SEQ ID NO:13 (i.e., CARTRSRSVRNCTSATCPVDAFDLW), respectively; and light chain variable region, the amino acid sequence of which has LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:14 (i.e., RSSQSLLRSNGYNYLD), SEQ ID NO:15 (i.e., LGSNRAS) and SEQ ID NO:16 (i.e., MQALQTPYT), respectively; or

[0033] (III) Heavy chain variable region, the amino acid sequence of which has HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:21 (i.e., TSGVSVG), SEQ ID NO:22 (i.e., LIYWDDDKRYSPSLTS) and SEQ ID NO:23 (i.e., PRYYGDSSGYYWI); and light chain variable region, the amino acid sequence of which has LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:24 (i.e., SGDALPKQYAY), SEQ ID NO:25 (i.e., KTSERPS) and SEQ ID NO:26 (i.e., QSADSSGFYV).

[0034] In some embodiments, the human neutralizing antibody or its antigen-binding fragment includes a heavy chain variable region and / or a light chain variable region.

[0035] (I) The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:7, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:7; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:8, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:8; or

[0036] (II) The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:17, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:17; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:18, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:18; or

[0037] (III) The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:27, or an amino acid sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:27; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:28, or an amino acid sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with the amino acid sequence shown in SEQ ID NO:28.

[0038] In some embodiments, the human neutralizing antibody or its antigen-binding fragment includes a heavy chain variable region and / or a light chain variable region.

[0039] (I) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:7, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:8. An antibody having the amino acid sequences shown in SEQ ID NO:7 and SEQ ID NO:8, respectively, is named YB9-258; or

[0040] (II) The heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO:17, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:18. An antibody having the amino acid sequences shown in SEQ ID NO:17 and SEQ ID NO:18, respectively, is named YB13-292; or

[0041] (III) The heavy chain variable region contains the amino acid sequence shown in SEQ ID NO:27; and the light chain variable region contains the amino acid sequence shown in SEQ ID NO:28. The antibody having the amino acid sequences shown in SEQ ID NO:27 and SEQ ID NO:28 respectively is named YB13-208.

[0042] In the above embodiments, the amino acid sequence of the heavy chain variable region of neutralizing antibody YB9-258, represented by SEQ ID NO:7, is as follows:

[0043] The underlined parts represent the framework regions FR1-4 of the heavy chain variable region, while the bolded parts represent HCDR1, HCDR2, and HCDR3 of the heavy chain variable region.

[0044] In the above embodiments, the amino acid sequence of the light chain variable region of neutralizing antibody YB9-258, as shown in SEQ ID NO:8, is as follows:

[0045] The underlined parts represent the framework regions FR1-4 of the light chain variable region, while the bolded parts represent the LCDR1, LCDR2, and LCDR3 of the heavy chain variable region.

[0046] In the above embodiments, the amino acid sequence of the heavy chain variable region of neutralizing antibody YB13-292, represented by SEQ ID NO:17, is as follows:

[0047] The underlined parts represent the framework regions FR1-4 of the heavy chain variable region, while the bolded parts represent HCDR1, HCDR2, and HCDR3 of the heavy chain variable region.

[0048] In the above embodiments, the amino acid sequence of the light chain variable region of neutralizing antibody YB13-292, represented by SEQ ID NO:18, is as follows:

[0049] The underlined parts represent the framework regions FR1-4 of the light chain variable region, while the bolded parts represent the LCDR1, LCDR2, and LCDR3 of the heavy chain variable region.

[0050] In the above embodiments, the amino acid sequence of the heavy chain variable region of neutralizing antibody YB13-208, represented by SEQ ID NO:27, is as follows:

[0051] The underlined parts represent the framework regions FR1-4 of the heavy chain variable region, while the bolded parts represent HCDR1, HCDR2, and HCDR3 of the heavy chain variable region.

[0052] In the above embodiments, the amino acid sequence of the light chain variable region of neutralizing antibody YB13-208, represented by SEQ ID NO:28, is as follows:

[0053] The underlined parts represent the framework regions FR1-4 of the light chain variable region, while the bolded parts represent the LCDR1, LCDR2, and LCDR3 of the heavy chain variable region.

[0054] In some embodiments, the human neutralizing antibody or its antigen-binding fragment is selected from Fab, Fab', Fab'-SH, Fv, scFv, F(ab')2, and biantibody.

[0055] In a second aspect, the present invention provides a polypeptide comprising a sequence selected from SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:27 and / or SEQ ID NO:28; optionally, the polypeptide of the second aspect comprises a sequence shown in SEQ ID NO:7 and / or SEQ ID NO:8; optionally, the polypeptide of the second aspect comprises a sequence shown in SEQ ID NO:17 and / or SEQ ID NO:18; optionally, the polypeptide of the second aspect comprises a sequence shown in SEQ ID NO:27 and / or SEQ ID NO:28.

[0056] Thirdly, a polynucleotide is provided that encodes the human neutralizing antibody or antigen-binding fragment thereof that binds to the coronavirus S protein.

[0057] Furthermore, the polynucleotide is DNA or mRNA.

[0058] Further, the polynucleotide has a nucleotide sequence such as SEQ ID NO:9 and / or SEQ ID NO:10; or the polynucleotide has a nucleotide sequence such as SEQ ID NO:19 and / or SEQ ID NO:20; or the polynucleotide has a nucleotide sequence such as SEQ ID NO:29 and / or SEQ ID NO:30.

[0059] In some embodiments, the polynucleotide has nucleotide sequences such as SEQ ID NO:9 and SEQ ID NO:10; or the polynucleotide has nucleotide sequences such as SEQ ID NO:19 and SEQ ID NO:20; or the polynucleotide has nucleotide sequences such as SEQ ID NO:29 and SEQ ID NO:30.

[0060] The nucleotide sequence shown in SEQ ID NO:9 corresponds to the amino acid sequence encoding SEQ ID NO:7. The nucleotide sequence shown in SEQ ID NO:9 is as follows:

[0061]

[0062] The nucleotide sequence shown in SEQ ID NO:10 corresponds to the amino acid sequence encoding SEQ ID NO:8. The nucleotide sequence shown in SEQ ID NO:10 is as follows:

[0063]

[0064] The nucleotide sequence shown in SEQ ID NO:19 corresponds to the amino acid sequence encoding SEQ ID NO:17. The nucleotide sequence shown in SEQ ID NO:19 is as follows:

[0065]

[0066] The nucleotide sequence shown in SEQ ID NO:20 corresponds to the amino acid sequence encoding SEQ ID NO:18. The nucleotide sequence shown in SEQ ID NO:20 is as follows:

[0067]

[0068] The nucleotide sequence shown in SEQ ID NO:29 corresponds to the amino acid sequence encoding SEQ ID NO:27. The nucleotide sequence shown in SEQ ID NO:19 is as follows:

[0069]

[0070]

[0071] The nucleotide sequence shown in SEQ ID NO:30 corresponds to the amino acid sequence encoding SEQ ID NO:28. The nucleotide sequence shown in SEQ ID NO:20 is as follows:

[0072]

[0073] Fourthly, the present invention provides a nucleic acid construct comprising the aforementioned polynucleotide.

[0074] More preferably, the polynucleotide further comprises at least one expression regulatory element operatively linked to the polynucleotide, such as a histidine tag, a stop codon, etc.

[0075] Fifthly, the present invention provides an expression vector comprising the aforementioned nucleic acid construct.

[0076] In a sixth aspect, the present invention provides a transformed cell comprising a polypeptide as described in the second aspect above, a polynucleotide as described in the third aspect above, a nucleic acid construct as described in the fourth aspect above, or an expression vector as described in the fifth aspect above. Optionally, it is a eukaryotic expression vector.

[0077] In another aspect, the present invention provides a pharmaceutical composition comprising a human neutralizing antibody or antigen-binding fragment thereof that binds to the coronavirus S protein as described in the first aspect above, a polypeptide as described in the second aspect above, a polynucleotide as described in the third aspect above, a nucleic acid construct as described in the fourth aspect above, an expression vector as described in the fifth aspect above, or transformed cells as described in the sixth aspect above, and a pharmaceutically acceptable carrier and / or excipient.

[0078] More preferably, the pharmaceutical composition is in the form of a nasal spray, oral formulation, suppository, or parenteral formulation.

[0079] More preferably, the nasal spray is selected from aerosols, sprays, and powders.

[0080] More preferably, the oral preparation is selected from tablets, powders, pills, granules, fine granules, soft / hard capsules, film-coated preparations, pellets, sublingual tablets, and ointments.

[0081] More preferably, the parenteral preparation is a transdermal preparation, ointment, plaster, topical liquid, injectable or bolus-applied preparation.

[0082] Eighthly, the present invention provides the use of a human neutralizing antibody or antigen-binding fragment thereof that binds to the coronavirus S protein as described in the first aspect above, a polypeptide as described in the second aspect above, a polynucleotide as described in the third aspect above, a nucleic acid construct as described in the fourth aspect above, an expression vector as described in the fifth aspect above, or a transformed cell as described in the sixth aspect above, or a pharmaceutical composition as described in the seventh aspect above, in the preparation of a medicament for the prevention or treatment, detection, or diagnosis of COVID-19 infection.

[0083] Preferably, the novel coronavirus is the original SARS-CoV-2 strain and / or a SARS-CoV-2 variant strain.

[0084] More preferably, the SARS-CoV-2 variant strains include Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Lambda (C.37), Kappa (B.1.617.1), Delta (B.1.617.2) and / or Omicron (B.1.1.529) strains.

[0085] Among them, neutralizing antibodies YB9-258 and YB13-292 showed good neutralizing activity against the original (WT) strain of SARS-CoV-2 and its variant strains Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Lambda (C.37), Kappa (B.1.617.1), Delta (B.1.617.2) and / or Omicron (B.1.1.529).

[0086] Neutralizing antibody YB13-208 exhibits good neutralizing activity against the original (WT) and variant strains of SARS-CoV-2, including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Lambda (C.37), Kappa (B.1.617.1), and / or Delta (B.1.617.2).

[0087] In a ninth aspect, the present invention provides a method for preventing or treating COVID-19, comprising: administering to a subject in need a preventive or therapeutically effective amount of a human neutralizing antibody or antigen-binding fragment thereof that binds to the coronavirus S protein as described in the first aspect above, a polypeptide as described in the second aspect above, a polynucleotide as described in the third aspect above, a nucleic acid construct as described in the fourth aspect above, an expression vector as described in the fifth aspect above, or a transformed cell as described in the sixth aspect above, or a pharmaceutical composition as described in the seventh aspect above.

[0088] Preferably, the novel coronavirus is the original SARS-CoV-2 strain and / or a SARS-CoV-2 variant strain.

[0089] More preferably, the SARS-CoV-2 variant strain is an Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Lambda (C.37), Kappa (B.1.617.1), Delta (B.1.617.2) and / or Omicron (B.1.1.529) strain.

[0090] The dosage of the active ingredient in the pharmaceutical composition of the present invention varies depending on the target patient, the target organ, symptoms, method of administration, etc. It can be determined based on the doctor's judgment, taking into account the type of dosage form, method of administration, patient's age and weight, patient's symptoms, etc.

[0091] Beneficial effects

[0092] (1) The neutralizing antibodies YB9-258 and YB13-292 of this invention exhibit good neutralizing activity against the original SARS-CoV-2 strain (WT), variant strains Alpha (B.1.1.7), Beta (B.1.351), Delta (B.1.617.2), Gamma (P.1), Lambda (C.37), and Omicron (B.1.1.529). The neutralizing antibodies YB9-258 and YB13-292 of this invention show high neutralizing activity against both true and false viruses of the highly pathogenic SARS-CoV-2 variant strains Delta (B.1.617.2) and Omicron (B.1.1.529), with IC50 values ​​of [missing information]. 50 ≤0.04ug / ml; The neutralizing antibody YB13-208 of this invention exhibits good neutralizing activity against the original SARS-CoV-2 strain (WT), variant strain Alpha (B.1.1.7), variant strain Beta (B.1.351), variant strain Delta (B.1.617.2), variant strain Gamma (P.1), variant strain Kappa (B.1.617.1), variant strain Delta (B.1.617.2), and variant strain Lambda (C.37), but has no neutralizing activity against the SARS-CoV-2 variant strain Omicron (B.1.1.529). The neutralizing antibodies YB9-258, YB13-292, and YB13-208 of this invention can effectively inhibit SARS-CoV-2 pseudovirus infection and also exhibit good neutralizing activity against the true virus. The neutralizing antibodies of this invention have clinical application value in the treatment and prevention of SARS-CoV-2 infection.

[0093] (2) The neutralizing antibodies obtained by screening in this invention have broad-spectrum neutralizing activity and can effectively neutralize SARS-CoV-2 and several highly infectious and harmful SARS-CoV-2 novel coronavirus variants, such as the UK's SARS-CoV-2 variant strain Alpha (B.1.1.7), the South African SARS-CoV-2 variant strain Beta (B.1.351), the Indian SARS-CoV-2 variant strain Delta (B.1.617.2), and the SARS-CoV-2 variant strain Omicron (B.1.1.529).

[0094] (2) This invention utilizes plasma from recovered patients infected with different SARS-CoV-2 variants to obtain broad-spectrum candidate antibodies. By employing flow cytometry and single-cell high-throughput assays, compared to traditional methods, the large-scale data obtained from high-throughput scVDJ sequencing (scVDJ-seq) allows for the rapid, efficient, and high-throughput detection of B-cell clonal enrichment before in vitro antibody expression. The stable and efficient eukaryotic expression system used in this invention ensures that the obtained antibodies are fully humanized, eliminating the need for further humanization.

[0095] (3) This invention collected plasma from recovered patients infected with different SARS-CoV-2 variants. First, B cells were enriched using magnetic beads, and SARS-CoV-2 S1-specific single B cells were sorted using flow cytometry. Single-cell library construction, BCR immune repertoire, and high-throughput transcriptome sequencing were performed using 10×Genomics technology. Through data analysis, antibody sequences were sorted and scored according to different parameters such as antibody sequence abundance, frequency, germline genes used, and mutation rate. Antibody sequences with potential affinity were selected for batch synthesis and eukaryotic expression and purification. Next, ELISA and SPR methods were used to test the binding and affinity of candidate antibodies to the S1 protein of different SARS-CoV-2 variants. The neutralizing abilities of different SARS-CoV-2 variants against pseudoviruses and true viruses were also investigated to ensure that the selected neutralizing antibodies were effective and broad-spectrum.

[0096] (4) This invention provides potential new antibody drugs for the clinical prevention, treatment and detection of the novel coronavirus. Attached Figure Description

[0097] One or more embodiments are illustrated by way of example with reference to the accompanying drawings, and these illustrative examples are not intended to limit the embodiments. The term "illustrative" as used herein means "serving as an example, embodiment, or illustration." Any embodiment illustrated herein as "illustrative" is not necessarily to be construed as superior to or better than other embodiments.

[0098] Figure 1Example 3 of this invention uses ELISA to analyze the binding activity of neutralizing antibodies against the S1 antigen of different original and variant strains of the novel coronavirus.

[0099] Figure 2 This is a schematic diagram showing the affinity constants of different neutralizing antibodies to the S1 antigen of different SARS-CoV-2 virus strains in Example 4 of the present invention;

[0100] Figure 3 This is a schematic diagram illustrating the neutralizing effect of the neutralizing antibody in Example 7 of the present invention on different SARS-CoV-2 pseudovirus strains. Detailed Implementation

[0101] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprising of," etc., will be understood to include the stated elements or components, and does not exclude other elements or other components.

[0102] Furthermore, to better illustrate the present invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In some embodiments, materials, elements, methods, and means well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0103] The present invention will now be described in detail.

[0104] definition

[0105] When referring to ligand / receptor, antibody / antigen, or other binding pairs, "specific" binding means determining the presence of the protein, for example, the binding reaction of the neutralizing antibody of this invention to the SARS-CoV-2S1 protein, within a heterogeneous population of proteins and / or other biological reagents. Therefore, under specified conditions, a particular ligand / antigen binds to a specific receptor / antibody and does not bind in significant amounts to other proteins present in the sample.

[0106] The reagents, enzymes, culture media, antibiotics, and milk used in the following examples of the present invention are all commercially available products. For example, the Chromium Single Cell V(D)J Reagent Kits (10X Genomics) kit was purchased from 10X Genomics.

[0107] The antigens and pseudoviruses used in the following embodiments of the present invention are all commercially available products. For example, the RBD antigen or S1 antigen was purchased from Beijing Sinocare Medical Technology Co., Ltd.

[0108] The original strain (WT)S1 antigen has the catalog number 40591-V08H and is manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0109] The original strain (WT) RBD antigen has the catalog number 40592-V08H and is manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0110] The catalog number of the variant strain Delta S1 antigen is 40591-V08H23, manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0111] The product code for the variant strain Delta RBD antigen is 40592-V08H90, manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0112] The catalog number for the variant strain Beta RBD antigen is 40592-V08H85, manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0113] The catalog number of the variant strain Beta S1 antigen is 40591-V08H10, manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0114] The variant strain Omicron RBD antigen has the catalog number 40592-V08H121 and is manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0115] The catalog number of the variant strain Omicron S1 antigen is 40591-V08H41, manufactured by Beijing Yiqiao Shenzhou Technology Co., Ltd.

[0116] The pseudovirus variant Omicron was purchased from Nanjing Novizan Biotechnology Co., Ltd., product number: DD1768-03.

[0117] The pseudovirus variant Delta was purchased from Suzhou Bio-Long Technology Co., Ltd., product number: BDAA0115.

[0118] The original pseudovirus strain WT was purchased from Nanjing Novizan Biotechnology Co., Ltd., product number: DD1402-03.

[0119] Some commonly used biological materials, such as competent cells, vectors, helper phages, and cells to be transformed, are also commercially available products.

[0120] The antibody numbers in the following examples correspond to:

[0121] Antibody YB9-258 has the amino acid sequence of the heavy chain variable region as shown in SEQ ID NO:7 and the amino acid sequence of the light chain variable region as shown in SEQ ID NO:8.

[0122] Antibody YB13-292 has the amino acid sequence of the heavy chain variable region as shown in SEQ ID NO:17 and the amino acid sequence of the light chain variable region as shown in SEQ ID NO:18.

[0123] Antibody YB13-208 has the amino acid sequence of the heavy chain variable region as shown in SEQ ID NO:27 and the amino acid sequence of the light chain variable region as shown in SEQ ID NO:28.

[0124] Example 1: Obtaining antibody sequence information through high-throughput single-cell sequencing

[0125] Blood was collected from patients who had recovered from infection with the Delta (B.1.617.2) variant of SARS-CoV-2. Peripheral blood mononuclear cells (PBMCs) were separated from the fresh blood using Ficoll gradient centrifugation. B cells were then isolated from the PBMCs using magnetic beads, and antigen-binding B cells were enriched. Library construction was then performed in a P2 laboratory according to the kit instructions (Chromium Single Cell V(D)J Reagent Kits, 10X Genomics). Sequencing was performed on the BGI sequencing platform using the MGISEQ-2000RS Sequencing Flow Cell Kit (manufactured by BGI Genomics). After sequencing, the data were analyzed, and antibody assembly was performed.

[0126] Example 2 Antibody protein expression and purification

[0127] We commissioned GenScript to perform antibody protein expression and purification.

[0128] Example 3: ELISA analysis of the binding ability of neutralizing antibodies to antigens of original and mutant SARS-CoV-2 strains.

[0129] Coat an ELISA plate with 50 ng of SARS-CoV-2 RBD antigen or SARS-CoV-2 S1 antigen and incubate overnight at 4°C. Wash three times with PBST (0.05%). Add 100 μL of 5% skim milk powder to each well and block at room temperature for 2 h. Wash three times with PBST. Serially dilute the neutralizing antibody with skim milk powder, starting at 1.5 μg / ml (10 nM), and then perform 5-fold serial dilutions for a total of 8 concentration gradients. Add each gradient to three wells, repeating for each gradient. Add 50 μL of primary antibody prepared with skim milk powder to a concentration of 2 μg / ml and incubate at room temperature for 1 h. Wash three times with PBST. Add 50 μL of 8000-fold diluted Anti-human-FC secondary antibody and incubate at room temperature for 45 min. Wash three times with PBST. Add 100 μL of TMB chromogenic buffer (abcam) and develop for 10 min. Add an equal volume of TMB stop buffer (abcam) to stop the development. Read the OD450 value. The binding ability of neutralizing antibodies to antigens of different SARS-CoV-2 strains was analyzed.

[0130] ELISA detection results using S1 antigen of different SARS-CoV-2 SARS-CoV-2 strains are as follows Figure 1 The results show that:

[0131] Neutralizing antibodies YB9-258 and YB13-292 exhibited good binding activity to the S1 antigen of the original SARS-CoV-2 strain (WT), the S1 antigen of the variant strain Alpha (B.1.1.7), the S1 antigen of the variant strain Beta (B.1.351), the S1 antigen of the variant strain Delta (B.1.617.2), the S1 antigen of the variant strain Gamma (P.1), the S1 antigen of the variant strain Lambda (C.37), and the S1 antigen of the variant strain Omicron (B.1.1.529), demonstrating good broad-spectrum binding activity.

[0132] Neutralizing antibody YB13-208 exhibits good binding activity against the S1 protein of the SARS-CoV-2 variant Omicron (B.1.1.529), except for low binding activity against the S1 antigen of other SARS-CoV-2 virus strains, and also has good broad-spectrum activity.

[0133] Example 4: Determination of the affinity constants of neutralizing antibodies to the original strain and mutant antigens using the SPR method.

[0134] The original SARS-CoV-2 strain (WT) S1 antigen, Delta (B.1.617.2) S1 antigen, and Omicron (B.1.1.529) S1 antigen were coupled to a CM5 chip. Antibody proteins were diluted with HBS-P buffer. In this example, 19.2 μg / ml (128 nM) was used as the initial antibody concentration, followed by sequential 2-fold serial dilutions for six concentration gradients, with the buffer used as a control. Each injection lasted 120 s, followed by 240 s of dissociation at a flow rate of 30 μL / min, and then regenerated with 10 mM pH 2.0 Glycine. This cycle was repeated until all concentration gradients were injected. After the program ran, the built-in analysis program of the Biacore T200 (GE) instrument was used for fitting analysis to obtain the neutralizing antibody affinity constant. The results are shown in Table 1 and [Table data missing]. Figure 2 The results showed that neutralizing antibodies YB9-258, YB13-292, and YB13-208 all had high affinity for the SARS-CoV-2 S1 antigen of the original strain (WT), the variant strain Delta (B.1.617.2), and the variant strain Omicron (B.1.1.529) of the novel coronavirus.

[0135] Table 1 shows the bond affinity and dissociation of each antibody and its constituent molecules, along with the dissociation constants KD(M).

[0136] Antibody information WT Delta Omicron YB9-258 7.17E-11 8.33E-10 4.17E-12 YB13-292 1.80E-10 3.91E-12 6.06E-09 YB13-208 8.20E-11 6.39E-10 3.11E-09

[0137] Example 5: Predicting Neutralizing Antibody Binding Competitive Epitopes Using the SPR Method

[0138] A CM5 chip was coated with the SARS-CoV-2 novel coronavirus Delta(B.1.617.2)S1 antigen until the response value reached 150 RU. Neutralizing antibodies were prepared at 200 nM using HBS-P buffer, loaded at a rate of 10 μL / min, with a saturation time of 200 s and a regeneration time of 60 s. Each antibody was saturated individually first, and the response value at saturation was recorded. In the competition experiment, one antibody was loaded and saturated first, followed by the other antibody. The response value and the trend of the real-time curve were observed to determine the epitope relationship between the two antibodies. If the two antibodies have different epitopes, the RU values ​​of both antibodies after loading will be the same as the response values ​​when saturated individually. If epitope competition exists, the value will decrease when the second antibody is loaded. The degree of decrease indicates the competition between the two antibodies. The results showed that these neutralizing antibodies were divided into two different epitopes. Neutralizing antibody YB9-258 had the same epitope as antibody YB13-208, while neutralizing antibody YB13-292 had a different epitope from both YB9-258 and YB13-208. This indicates that neutralizing antibody YB13-292 can be used in combination with neutralizing antibodies YB9-258 or YB13-208.

[0139] Example 6: Detection of antibody neutralization effect using a COVID-19 pseudovirus experiment.

[0140] The serum samples to be tested were inactivated at 56°C for 30 min, centrifuged at 6000g for 3 min, and the supernatant was transferred to 2.5 ml centrifuge tubes for later use. 120 μl / well of DMEM serum-free medium was added to B11-D11 (cell control CC, see Table 2); 60 μl / well of DMEM serum-free medium was added to E11-G11 (virus control VC, see Table 2); 84 μl / well of DMEM serum-free medium was added to G2-G10; and 60 μl / well of DMEM serum-free medium was added to B2-F10. Serum was diluted 30-fold, with serum sample 1 (6 μl) added to wells G2, G3, and G4, and so on, with serum sample 4 (6 μl) added to wells G8, G9, and G10. Each serum sample was then diluted 3-fold. Set the multichannel pipette to 30 μl. Gently pipette the liquid in wells G2-G4 6-8 times to mix thoroughly. Then transfer 30 μl of liquid to the corresponding wells F2-F4, gently pipette 6-8 times, and then transfer to wells E2-E4, and so on. Finally, discard 30 μl of liquid from wells B2-B4. Refer to the table for the sample addition order. Perform this procedure for other samples on the plate. Dilute the pseudovirus to 1.3 × 10⁻⁶ using DMEM serum-free medium. 4 (1×10 4 ~2×10 4 Add 50 μl of TCID50 / ml to columns 2-10 and wells E11-G11 (virus control VC, see Table 2) to make the amount of pseudovirus per well 650 (500-1000) / well. Incubate the 96-well plate in a cell culture incubator (37°C, 5% CO2) for 1 hour. When the incubation time reaches half an hour, remove the pre-prepared 293T-hACE2 cells (with a confluence of 80%–90%) from the incubator. Using a T75 culture flask as an example, aspirate the culture medium, add 5 ml of PBS buffer to wash the cells, discard the PBS, add 3 ml of 0.25% trypsin-EDTA, and immerse the cells for 1 minute for digestion. Discard the trypsin, place the flask in a cell culture incubator for 5 minutes of digestion, gently tap the side of the flask to detach the cells, add 10 ml of culture medium to neutralize the trypsin, pipette a few times, transfer to a centrifuge tube, centrifuge at 210g for 5 minutes, discard the supernatant, resuspend the cells in 10 ml of DMEM complete culture medium, count the cells, and dilute the cells to 2 × 10⁻⁶ cells / mL with DMEM complete culture medium. 5 Cells / ml. Incubate for 1 hour, then use a multichannel pipette to transfer 100 μl of the sample pseudovirus mixture previously incubated on a clear plate to a TC-treated white plate. Add 100 μl of cells to each well of the 96-well white plate, making the cell density 2 × 10⁶ cells per well. 4Gently shake the 96-well plate back and forth and side to side to disperse the cells evenly in the wells. Place the 96-well plate in a cell culture incubator and incubate at 37°C and 5% CO2 for 72 hours. After 72 hours, remove the 96-well plate from the cell culture incubator, and use a multichannel pipette to aspirate 100 μl of supernatant from each sample well. Then add 100 μl of luciferase assay reagent and react at room temperature in the dark for 2 minutes. Calculate the neutralization inhibition rate: Inhibition rate = [1 - (mean luminescence intensity of sample group - mean CC value of blank control) / (mean luminescence intensity of negative group - mean CC value of blank control)] × 100%. Based on the neutralization inhibition rate results, calculate the IC50 using the Reed-Muench method. 50 .

[0141] The experimental results using the original SARS-CoV-2 strain (WT) pseudovirus, the variant Delta pseudovirus, and the variant Omicron pseudovirus are as follows: Figure 3 As shown, neutralizing antibodies YB9-258 and YB13-292 exhibited neutralizing activity against the original strain (WT), the variant strain Delta, and the variant strain Omicron pseudovirus. However, neutralizing antibody YB13-208 showed no or low neutralizing activity against the SARS-CoV-2 variant strain Omicron pseudovirus. Neutralizing antibodies YB9-258 and YB13-292 showed high neutralizing activity against the SARS-CoV-2 variant strain Omicron pseudovirus, with an IC50 value of [missing information]. 50 Less than 0.04 ug / ml indicates that they have a higher ability to inhibit the infection of target cells by the Omicron (B.1.1.529) variant strain.

[0142] Example 7: Detection of the neutralizing effect of antibodies against different evovirus strains

[0143] The neutralizing activity of neutralizing antibodies was tested by the Guangdong Provincial Center for Disease Control and Prevention, including the original strain (WT), the variant strain Delta (B.1.617.2), and the variant strain Omicron (B.1.1.529) of the virus. The results are shown in Table 2.

[0144] Table 2 shows that neutralizing antibodies YB9-258 and YB13-292 exhibited strong neutralizing ability against the original SARS-CoV-2 strain (WT), the variant Delta (B.1.617.2), and the variant Omicron (B.1.1.529) virus. Neutralizing antibody YB13-208 showed strong neutralizing ability against both the original SARS-CoV-2 strain (WT) and the Delta (B.1.617.2) variant, but no neutralizing ability against the SARS-CoV-2 variant Omicron (B.1.1.529) virus. The results in Table 2 also indicate a good correlation between the neutralizing activity of the antibodies and pseudoviruses, as well as their neutralizing activity against real viruses.

[0145] Table 2. Neutralizing effects of various neutralizing antibodies against different eukaryotes.

[0146]

[0147] In summary, neutralizing antibodies YB9-258, YB13-292, and YB13-208 can serve as neutralizing antibodies against the novel coronavirus (SARS-CoV-2) with high neutralizing activity. Among them, neutralizing antibodies YB9-258 and YB13-292 exhibit excellent broad-spectrum neutralization, while neutralizing antibody YB13-208 also shows good broad-spectrum neutralization.

[0148] 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 of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A human neutralizing antibody or antigen-binding fragment thereof that binds to the S protein of the coronavirus SARS-CoV-2, comprising a heavy chain variable region and a light chain variable region. The heavy chain variable region includes: The amino acid sequences are as shown in SEQ ID NO:21 for HCDR1, as shown in SEQ ID NO:22 for HCDR2, and as shown in SEQ ID NO:23 for HCDR3. The light chain variable region includes: The amino acid sequences are as shown in SEQ ID NO:24 for HCDR1, as shown in SEQ ID NO:25 for HCDR2, and as shown in SEQ ID NO:26 for HCDR3.

2. The human neutralizing antibody or its antigen-binding fragment according to claim 1, characterized in that, It contains variable regions for heavy chains and variable regions for light chains: The heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:27, or an amino acid sequence having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO:27; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:28, or an amino acid sequence having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO:

28.

3. The human neutralizing antibody or its antigen-binding fragment as described in claim 1 or 2, wherein the antigen-binding fragment is selected from Fab, Fab', Fab'-SH, Fv, scFv, F(ab')2.

4. An antibody that binds to the S protein of coronavirus SARS-CoV-2, having a heavy chain variable region as shown in SEQ ID NO:27 and a light chain variable region as shown in SEQ ID NO:

28.

5. A polynucleotide encoding the human neutralizing antibody or antigen-binding fragment thereof as described in any one of claims 1 to 3.

6. The polynucleotide according to claim 5, characterized in that, The polynucleotide is DNA or mRNA; the polynucleotide has a nucleotide sequence such as SEQ ID NO:29 and SEQ ID NO:

30.

7. A nucleic acid construct comprising the polynucleotide of claim 5 or 6.

8. An expression vector comprising the nucleic acid construct of claim 7.

9. A transformed cell comprising the polynucleotide of claim 5 or 6, the nucleic acid construct of claim 7, or the expression vector of claim 8.

10. A pharmaceutical composition comprising the human neutralizing antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 3, the antibody as claimed in claim 4, the polynucleotide as claimed in claim 5 or 6, the nucleic acid construct as claimed in claim 7, the expression vector as claimed in claim 8 or the transformed cell as claimed in claim 9, and a pharmaceutically acceptable carrier and / or excipient.

11. The pharmaceutical composition according to claim 10, characterized in that, The pharmaceutical composition is a nasal spray, an oral preparation, or a suppository.

12. The pharmaceutical composition according to claim 10, characterized in that, The pharmaceutical composition is in the form of a parenteral preparation.

13. The use of a human neutralizing antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, an antibody according to claim 4, a polynucleotide according to claim 5 or 6, a nucleic acid construct according to claim 7, an expression vector according to claim 8, a transformed cell according to claim 9, or a pharmaceutical composition according to any one of claims 10 to 12 in the preparation of a medicament for the prevention or treatment or detection or diagnosis of COVID-19 infection; The novel coronavirus mentioned refers to both the original SARS-CoV-2 strain and the SARS-CoV-2 variant strain; The SARS-CoV-2 variants mentioned are Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Kappa (B.1.617.1), Lambda (C.37), and Delta (B.1.617.2).

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