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Single chain antibody capable of resisting S2 protein on surface of novel coronavirus SARS-CoV-2 and applications thereof

A single-chain antibody, sars-cov-2 technology, applied in the field of cellular immunology and molecular virology, can solve the problems of listing, complicated virus prevention and control, no effective therapeutic drugs and vaccines, etc., and achieve the effect of inhibiting infection

Active Publication Date: 2021-07-23
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These factors make the prevention and control of the virus very complicated, and there are currently no effective treatments and vaccines on the market

Method used

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  • Single chain antibody capable of resisting S2 protein on surface of novel coronavirus SARS-CoV-2 and applications thereof
  • Single chain antibody capable of resisting S2 protein on surface of novel coronavirus SARS-CoV-2 and applications thereof
  • Single chain antibody capable of resisting S2 protein on surface of novel coronavirus SARS-CoV-2 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Single-chain antibody scFv-27 sequence

[0056] 1. Antibody screening

[0057] Phage scFv display library construction

[0058] 1. Collect peripheral blood from convalescent patients with COVID-19, and isolate B cells from peripheral blood

[0059] The experiment was conducted on May 19, 2020. After informed consent, 20ml of peripheral blood was collected from 20 convalescent patients diagnosed with COVID-19. B cells in 20ml heparin anticoagulated blood were separated by density gradient centrifugation.

[0060] 2. RNA extraction and cDNA synthesis in PBMC

[0061] Extract PBMC cellular RNA, and then use a synthesis kit to reverse transcribe the RNA into cDNA.

[0062] 3. PCR amplification of VK, VL and VH

[0063] (1) Amplify the VK&VL system, as shown in Table 1.

[0064] Table 1 Amplified VK&VL system

[0065] solution or component Volume (μL) cDNA 1 EX Buffer(10x) 4 dNTPs (10mM each) 5 P1 (10μM) 2 P2 (10μM) ...

Embodiment 2

[0110] Example 2 Single-chain antibody scFv-32 sequence

[0111] 1. Antibody screening

[0112] Phage scFv display library construction

[0113] 1. Collect peripheral blood from convalescent patients with COVID-19, and isolate B cells from peripheral blood

[0114] The experiment was conducted on May 19, 2020. After informed consent, 20ml of peripheral blood was collected from 20 convalescent patients diagnosed with COVID-19. B cells in 20ml heparin anticoagulated blood were separated by density gradient centrifugation.

[0115] 2. RNA extraction and cDNA synthesis in PBMC

[0116] Extract PBMC cellular RNA, and then use a synthesis kit to reverse transcribe the RNA into cDNA.

[0117] 3. PCR amplification of VK, VL and VH

[0118] (1) Amplify the VK&VL system, as shown in Table 1.

[0119] Table 1 Amplified VK&VL system

[0120]

[0121]

[0122] (2) A system for amplifying the Fd segment of the heavy chain, as shown in Table 2.

[0123] Table 2 Amplified heavy ...

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Abstract

The invention provides a single chain antibody capable of resisting S2 protein on the surface of novel coronavirus SARS-CoV-2, and relates to the technical field of cellular immunology and molecular virology. The monoclonal antibody includes a variable region of a heavy chain of an scFv-27 sequence including amino acid sequences shown as SEQIDNO. 1-3, and a variable region of a light chain including amino acid sequences shown as SEQIDNO. 5-7, or include a variable region of a heavy chain of an scFv-32 including the amino acid sequences shown as SEQIDNO. 9-11, and a variable region of a light chain including the amino acid sequences shown as SEQIDNO. 13-16. The single chain antibody can be specifically combined with the S2 protein of the novel coronavirus SARS-CoV-2; and the single chain antibody has neutralization activity, can effectively inhibit infection of the novel coronavirus to target cells, and can be used for developing drugs for preventing or treating the infection of the novel coronavirus.

Description

technical field [0001] The invention relates to the technical fields of cellular immunology and molecular virology, in particular to a single-chain antibody against the S2 protein on the surface of the new coronavirus SARS-CoV-2 and its application. Background technique [0002] With the in-depth understanding of the epidemic situation from all walks of life, the follow-up is to study various detection reagents. At the current stage of the epidemic, all walks of life pay more attention to the vaccine and antiviral drug industry, and the research and development of antibody drugs will definitely become a mid-to-long-term trend. [0003] After 30 years of development, antibody drugs have become an important part of the global pharmaceutical market. The antibody drug market is still dominated by tumor and autoimmunity. With the continuous deepening of people's understanding of diseases and the continuous evolution of antibody technology, antibody drugs have gradually penetrate...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/46C12N15/13G01N33/569A61K39/42A61P31/14
CPCC07K16/10C07K16/468G01N33/56983A61P31/14C07K2317/565C07K2317/622C07K2317/76C07K2317/31A61K2039/505G01N2333/165G01N2469/10Y02A50/30
Inventor 胡佳周清华许峰阴丽媛梁好
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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