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Monoclonal antibody capable of aiming at corona virus disease 2019 virus spike protein non-RBD (Receptor-Binding Domain), and application of monoclonal antibody

A monoclonal antibody and spike protein technology, applied in the field of bioengineering, can solve problems such as ineffective effects and side effects, and achieve the effects of simple and efficient screening, strong specificity, and high-efficiency expression

Active Publication Date: 2020-11-27
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no effective drug in the treatment of new coronary pneumonia, and the effect of conventional antibiotic drug treatment is not obvious, and some have serious side effects

Method used

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  • Monoclonal antibody capable of aiming at corona virus disease 2019 virus spike protein non-RBD (Receptor-Binding Domain), and application of monoclonal antibody
  • Monoclonal antibody capable of aiming at corona virus disease 2019 virus spike protein non-RBD (Receptor-Binding Domain), and application of monoclonal antibody
  • Monoclonal antibody capable of aiming at corona virus disease 2019 virus spike protein non-RBD (Receptor-Binding Domain), and application of monoclonal antibody

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Experimental program
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Effect test

Embodiment 1

[0037] 1. Amplification of phage display antibody library:

[0038] Inoculate 500 μL of Escherichia coli TG-1 containing Tomlinson I+J phage display antibody library (MRC HGMP Resource Center, UK) into 25 mL of 2YT medium (1.6% Tryptone, 1% Yeast Extract) containing 100 μg / mL ampicillin and 1% glucose , 0.5% NaCl), cultured at 37°C to OD 600 is 0.4, add 10 9cfu (Colony FormationUnit) KM13 helper phage (MRC HGMP Resource Center, UK), after infection for 1 hour at 37°C, centrifuge at 3000g for 30 minutes, discard the supernatant, and use bacteriophage containing 100μg / ml ampicillin, 50μg / ml kanamycin and 0.1% Glucose 2YT medium 50mL (1.6% Tryptone, 1% Yeast Extract, 0.5% NaCl) to suspend the bacteria, shake the bacteria at 250rpm at 30°C for 16 hours, centrifuge the culture solution at 5000g for 30 minutes the next day, separate and recover the supernatant 40mL, add 10mL of PEG / NaCl solution to the supernatant, mix well and place on ice for 30 minutes, centrifuge at 5000g for ...

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Abstract

The invention discloses a monoclonal antibody capable of aiming at a corona virus disease 2019 virus spike protein non-RBD (Receptor-Binding Domain), and application of the monoclonal antibody. The monoclonal antibody is subjected to specific binding with the corona virus disease 2019 virus spike protein non-RBD and comprises the complementary determining regions CDRH1, CDRH2 and CDRH3 of a variable region of heavy chain and the complementary determining regions CDRL1, CDRL2 and CDRL3 of a variable region of light chain. The monoclonal antibody capable of aiming at the corona virus disease 2019 virus spike protein non-RBD has a high titer and high specificity, can realize efficient expression and can carry out the specific binding with domains except the spike protein RBD on the surface ofthe SARS-CoV-2, is used for qualitatively and quantitatively detecting the corona virus disease 2019 virus, can neutralize certain corona virus disease 2019 virus virulence and weaken the corona virus disease 2019 virus virulence, and performs a function of preventing or / and treating the corona virus disease 2019 virus pneumonia.

Description

technical field [0001] The present invention relates to the technical field of bioengineering, specifically to the technical field of cellular immunology and molecular biology, and more specifically to a monoclonal antibody against the non-RBD region of the spike protein of the novel coronavirus and its application. Background technique [0002] The membrane proteins of the new coronavirus mainly include the surface spike protein (Spike protein, S protein) and membrane protein (Membrane protein, M protein). The S protein is a glycoprotein that mainly acts on cell adhesion and cell membrane fusion. In mammals, the S protein is decomposed into S1 and S2 by furin or other enzymes, wherein S1 has a receptor attachment site, and S2 mainly shows fusion activity. The novel coronavirus can bind to a variety of cell receptors, among which angiotensin-converting enzyme 2 (ACE2), as a peptidase, is one of the receptors of the cell surface coronavirus. There is a part of S1 tightly bou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/577G01N33/569A61K39/42A61P31/14A61P11/00
CPCA61K2039/505A61P11/00A61P31/14C07K16/10C07K2317/35C07K2317/56C07K2317/565C07K2317/76G01N33/56983G01N33/577G01N2333/165G01N2469/10
Inventor 董金华陈丽梅李海梅
Owner WEIFANG MEDICAL UNIV
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