Anti-novel coronavirus monoclonal antibody and application thereof

A monoclonal antibody, antibody technology, used in antiviral agents, antiviral immunoglobulins, antibodies, etc.

Active Publication Date: 2020-08-28
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no studies have reported that antibodies with high binding activity and / or high neutralizing activity against 2019-nCoV

Method used

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  • Anti-novel coronavirus monoclonal antibody and application thereof
  • Anti-novel coronavirus monoclonal antibody and application thereof
  • Anti-novel coronavirus monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1: Isolation of memory B cells

[0082] Collect the blood (provided by Beijing You'an Hospital) of people infected with SARS-CoV-2 virus and discharged after recovery, and use STEMCELL SepMate in the P2+ biosafety laboratory TM -15 (Stemcell Technologies, catalog number: 86415) for the extraction of PBMCs. Subsequently, the extracted PBMCs were enriched for memory B cells using the STEMCELL EasySep HumanMemory B Cell Isolation Kit (Stemcell Technologies, catalog number: 17864) according to the manufacturer's instructions.

Embodiment 2

[0083] Example 2: Obtaining and Identification of Antibody Sequences

[0084] According to the manufacturer's instructions, Chromium Single Cell V(D)J Reagent Kits (purchased from 10X genomics, catalog number: 100006) were used to perform single-cell transcriptome VDJ sequencing on the above-mentioned enriched memory B cells. After analyzing the sequencing results, an antibody strain was obtained, which was named BD23. The sequence information of the BD23 antibody is as follows:

[0085] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 7 (the encoding gene is shown in SEQ ID NO: 17), wherein the amino acid sequence of CDR1-3 of the heavy chain variable region is shown in SEQ ID NO: 1 -3 (its coding gene is shown in SEQ ID NO:11-13 respectively);

[0086] The amino acid sequence of the light chain variable region is shown in SEQ ID NO: 8 (the encoding gene is shown in SEQ ID NO: 18), wherein the amino acid sequence of CDR1-3 of the light chain...

Embodiment 3

[0089] Example 3: Preparation and purification of antibody BD23

[0090] According to the sequence information of the BD23 antibody identified in Example 2, Beijing Yiqiao Shenzhou Co., Ltd. was commissioned to express and purify the BD23 antibody, and to detect the antigen reactivity of the BD23 antibody.

[0091] Briefly, nucleic acid molecules encoding antibody heavy and light chains were synthesized in vitro, and then cloned into expression vectors, respectively, to obtain recombinant expression vectors encoding antibody heavy and light chains, respectively. HEK293 cells were co-transfected with the above-mentioned recombinant expression vectors encoding antibody heavy chain and light chain respectively. After 4-6 hours of transfection, the cell culture medium was replaced with serum-free medium, and cultured at 37°C for 6 days. After the culture is over, the antibody protein expressed by the cells is purified from the culture through an affinity purification column. Sub...

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Abstract

The invention relates to the fields of immunology and molecular virology, in particular to the fields of diagnosis, prevention and treatment of novel coronaviruses. Specifically, the present inventionrelates to an anti-novel coronavirus monoclonal antibody, and a composition (e.g., a diagnostic agent and a therapeutic agent) comprising the antibody. Furthermore, the invention also relates to an application of the antibody. The antibody provided by the invention can be applied to diagnosis, prevention and / or treatment of infection of the novel coronavirus and / or diseases (e.g., novel coronavirus pneumonia) caused by the infection.

Description

technical field [0001] The present invention relates to the field of immunology and molecular virology, especially the field of diagnosis, prevention and treatment of novel coronavirus. In particular, the present invention relates to monoclonal antibodies against novel coronaviruses, and compositions (eg, diagnostic and therapeutic agents) comprising said antibodies. Furthermore, the present invention also relates to the use of said antibody. The antibody of the present invention can be used for diagnosing, preventing and / or treating novel coronavirus infection and / or diseases caused by said infection (for example, novel coronavirus pneumonia). Background technique [0002] The novel coronavirus SARS-CoV-2 is the causative agent of novel coronavirus pneumonia (COVID-19). ) and the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) that caused the outbreak in 2012 belong to the Coronaviridae family. Coronavirus particles are round or oval, also pleomorphic, with a dia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/577G01N33/569A61K39/42A61K45/06A61P31/14
CPCC07K16/10G01N33/577G01N33/56983A61K39/42A61K45/06A61P31/14C07K2317/565C07K2317/55C07K2317/24C07K2317/54G01N2333/165A61K2039/505A61K2300/00G01N2469/10C07K2317/76C12N15/63
Inventor 谢晓亮曹云龙孙文洁
Owner PEKING UNIV
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