Single-chain antibody for resisting S1 protein on surface of new coronavirus SARS-CoV-2 and application of single-chain antibody

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

Active Publication Date: 2021-08-17
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 for resisting S1 protein on surface of new coronavirus SARS-CoV-2 and application of single-chain antibody
  • Single-chain antibody for resisting S1 protein on surface of new coronavirus SARS-CoV-2 and application of single-chain antibody
  • Single-chain antibody for resisting S1 protein on surface of new coronavirus SARS-CoV-2 and application of single-chain antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Single-chain antibody scFv-22 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]

[0066]

[0067] (2) A system for amplifying the Fd segment of the heavy chain, as shown in Table 2.

[0068] Table 2 Amplified heavy ...

Embodiment 2

[0111] Example 2 Single-chain antibody scFv-24 sequence

[0112] 1. Antibody screening

[0113] Phage scFv display library construction

[0114] 1. Collect peripheral blood from convalescent patients with COVID-19, and isolate B cells from peripheral blood

[0115] 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.

[0116] 2. RNA extraction and cDNA synthesis in PBMC

[0117] Extract PBMC cellular RNA, and then use a synthesis kit to reverse transcribe the RNA into cDNA.

[0118] 3. PCR amplification of VK, VL and VH

[0119] (1) Amplify the VK&VL system, as shown in Table 1.

[0120] Table 1 Amplified VK&VL system

[0121]

[0122]

[0123] (2) A system for amplifying the Fd segment of the heavy chain, as shown in Table 2.

[0124] Table 2 Amplified heavy ...

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PUM

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Abstract

The invention provides a single-chain antibody for resisting S1 protein on the surface of a new coronavirus SARS-CoV-2, and relates to the technical fields of cellular immunology and molecular virology. The monoclonal antibody comprises heavy-chain variable regions of an scFv-22 sequence and with amino acid sequences shown as SEQ ID NO. 1-3 and light-chain variable regions of the scFv-22 sequence and with amino acid sequences shown as SEQ ID NO. 5-7, or heavy-chain variable regions of a scFv-24 sequence and with amino acid sequences shown as SEQ ID NO. 9-11 and light-chain variable regions of tje scFv-24 sequence and with amino acid sequences shown as SEQ ID NO. 13-16. The single-chain antibody disclosed by the invention can specifically bind to S1 protein of the novel coronavirus SARS-CoV-2, has neutralizing activity on the novel coronavirus, effectively inhibits infection of the novel coronavirus on target cells, and can be used for developing drugs for preventing or treating novel coronavirus infection.

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 surface S1 protein of the new coronavirus SARS-CoV-2 and its application. Background technique [0002] The global spread of new coronary pneumonia has aroused the attention of the antiviral drug industry from all walks of life. In the Chinese market, protection concepts such as masks and disinfectants were initially sought after. 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 anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10G01N33/569C07K16/46
CPCC07K16/10G01N33/56983G01N2333/165C07K2317/31C07K2317/565C07K2317/56Y02A50/30
Inventor 胡佳周清华许峰阴丽媛梁好
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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