Single-chain antibody of SARS-CoV-2 virus as well as screening method and application of single-chain antibody

A single-chain antibody, sars-cov-2 technology, applied in the direction of antibodies, antiviral agents, antiviral immunoglobulin, etc., can solve the problems of difficult antibody screening and large randomness, and achieve good specificity

Pending Publication Date: 2022-04-26
AB&B BIO TECH CO LTD JS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For the SARS-CoV-2 virus, both pharmaceuticals and testing require antibodies with good specificity, but there are differences in the specificity of different antibodies, and the selection of antibodies with good specificity is of great significance for pharmaceuticals and testing, but the screening of antibodies It is difficult, the randomness is very large, and no antibody with good specificity has been screened out in the prior art

Method used

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  • Single-chain antibody of SARS-CoV-2 virus as well as screening method and application of single-chain antibody
  • Single-chain antibody of SARS-CoV-2 virus as well as screening method and application of single-chain antibody

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Embodiment

[0041] In order to detect the binding strength of the single-chain antibody to the antigen, the present invention has carried out a monoclonal ELISA, and the steps are as follows:

[0042] (1) Prepare monoclonal phage supernatant

[0043] In order to have a sufficient number of blank control groups, the present invention randomly picks 96 single clones from the plate for the second screening and determination of titer in a 96-well deep-well plate, adds 300ul of 2YT medium at 180rpm, and cultivates it at 37°C until In the logarithmic phase, M13K07 was added to assist phage infection, the medium was changed, and cultured overnight at 30°C.

[0044] (2) Prepare the target protein, and coat the target protein on the ELISA plate.

[0045] For antigen coating, the target protein was diluted to a concentration of 2ug / m with coating solution (PBS 9.0), and then added to a 96-well ELISA plate, 50ul / well, and left overnight at 4°C. Wash, block, and wash ELISA plates again.

[0046] (...

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Abstract

The invention discloses a single-chain antibody of an SARS-CoV-2 virus, the single-chain antibody is specifically bound with an antigen S1-RBD, the single-chain antibody has the function of being bound with the S1-RBD, the S1-RBD is prevented from being bound with a receptor ACE2 on a cell, and the single-chain antibody can be applied to development of a neutralizing antibody. The single-chain antibody disclosed by the invention has good specificity on the SARS-CoV-2 virus.

Description

technical field [0001] The present invention relates to a single-chain antibody of SARS-CoV-2 virus and its screening method and application, in particular to a single-chain antibody with good specificity for SARS-CoV-2 virus. Background technique [0002] Viruses of the genus Coronavirus are RNA viruses wrapped in a mantle, with a diameter of about 100-160 nm, and the genetic material is the largest among all RNA viruses. There are three glycoproteins on the surface of the fatty membrane that wraps the virion: the spike glycoprotein (S, is the receptor binding site, cytolysis and the main antigenic site); the small envelope glycoprotein (E, smaller, with the cell membrane binding protein); membrane glycoprotein (M, responsible for transmembrane transport of nutrients, release of nascent virus budding and formation of viral outer envelope). [0003] The S protein is hydrolyzed into S1 and S2, and a part of S1 is the receptor binding domain (RBD) tightly combined with ACE2, ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C07K16/10G01N33/569A61K39/395A61P31/14
CPCC07K16/10G01N33/56983A61P31/14G01N2333/165G01N2469/10A61K2039/505
Inventor吴浩飞徐奇杨靖葛灵睿谢小静汪雨航游洪伟贾兰淑汤静雯沈鹏飞
OwnerAB&B BIO TECH CO LTD JS