Monoclonal antibody against RBD region of novel coronavirus SARS-CoV-2 spike protein and application of monoclonal antibody

A monoclonal antibody, spike protein technology, applied in the direction of antiviral immunoglobulin, application, antibody, etc., can solve problems such as toxic side effects, no obvious therapeutic effect, etc., to achieve strong specificity, prevention or/and treatment of new coronavirus Pneumonia, the effect of high expression

Active Publication Date: 2020-11-27
WEIFANG MEDICAL UNIV
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Problems solved by technology

[0004] At present, there is no specific therapeutic drug or vaccine for the new coronavirus on the market, because the anti-new coronavirus drugs mainly include the small molecule drug Remdesivir, the combination of chloroquine and hydroxychloroquine, the combination of lopinavir and ritonavir, and the combination of lopinavir and lopinavir. The three-drug combination of navir, ritonavir and interferon has no obvious therapeutic effect, and some even have serious side effects

Method used

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  • Monoclonal antibody against RBD region of novel coronavirus SARS-CoV-2 spike protein and application of monoclonal antibody
  • Monoclonal antibody against RBD region of novel coronavirus SARS-CoV-2 spike protein and application of monoclonal antibody
  • Monoclonal antibody against RBD region of novel coronavirus SARS-CoV-2 spike protein and application of monoclonal antibody

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Embodiment 1

[0034] 1. Amplification of phage display antibody library:

[0035] 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) suspended bacteria, 30°C, 250rpm speed shake for 16 hours, the next day, 5000g, 30 minutes of centrifugation of the culture medium, separation and recovery of supernatant 40mL , add 10mL of PEG / NaCl solution to the supernatant, mix well, place on ice for 30 minutes, centrifuge at 5000g for 30 minutes, di...

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Abstract

The invention discloses a monoclonal antibody against a RBD region of novel coronavirus SARS-CoV-2 spike protein and application of the monoclonal antibody. The monoclonal antibody specifically bindsto the RBD region of the novel coronavirus SARS-CoV-2 spike glycoprotein. The monoclonal antibody comprises complementarity determining regions of CDRH1, CDRH2, CDRH3 of a heavy chain variable regionand complementarity determining regions of CDRL1, CDRL2, CDRL3 of a light chain variable region. According to the monoclonal antibody against the RBD region of the novel coronavirus SARS-CoV-2 spike protein, the titer is high, the specificity is higher, efficient expressing can be achieved, the monoclonal antibody can specifically bind to the RBD region of the spike protein on the surface of the novel coronavirus SARS-CoV-2 and can be used for the detection of the ovel coronavirus SARS-CoV-2, certain toxicity of the novel coronavirus can be neutralized and reduced, and the effect of preventingor / and treating the novel coronavirus pneumonia is achieved.

Description

technical field [0001] The invention relates to the technical field of biotechnology, in particular to cellular immunology and molecular biology technology, specifically to a monoclonal antibody against the RBD region of the spike protein of the novel coronavirus SARS-CoV-2 and its application. Background technique [0002] The new coronavirus SARS-CoV-2 is a single-stranded positive-sense RNA virus, and its genome encodes four structural proteins: spike protein (S), small protein (E), matrix protein (M) and nucleocapsid protein (N ). The S protein is a type I fusion protein that forms a trimer on the surface of the virus particle and consists of two subunits, S1 and S2: S1 is responsible for receptor binding, and S2 is responsible for membrane fusion. The receptor binding domain (RBD) in S1 interacts with the human SARS-CoV receptor angiotensin-converting enzyme II (ACE2) molecule, while S2 contains the basic elements required for the membrane fusion process to realize the...

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 董金华李海梅陈丽梅
Owner WEIFANG MEDICAL UNIV
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