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

A monoclonal antibody and spike protein technology, applied in the fields of cellular immunology and molecular biology, can solve the problem of no new coronavirus treatment drugs or vaccines being marketed, and achieve the prevention or/and treatment of new coronavirus pneumonia, high-efficiency expression, specific effect

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

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Problems solved by technology

[0005] Various countries have proposed a variety of treatment options for the treatment of new coronary pneumonia, but currently there is no specific drug or vaccine for the new coronavirus on the market

Method used

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

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Embodiment 1

[0036] 1. Amplification of phage display antibody library:

[0037]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 9 cfu (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 a mixture containing 100 μg / mL ampicillin, 50 μg / mL kanamycin and 0.1% Glucose 2YT medium 50mL (1.6% Tryptone, 1% Yeast Extract, 0.5% NaCl) suspend the bacteria, shake the bacteria at 30°C and 250rpm 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, place on ice for 30 minutes, centrifuge at 5000g for 30 min...

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Abstract

The invention discloses a monoclonal antibody for a novel coronavirus virus SARS-CoV-2 spike protein RBD region and an application of the monoclonal antibody, and belongs to the field of cell immunology and the like. The monoclonal antibody is specifically combined with a novel coronavirus virus SARS-CoV-2 spike glycoprotein RBD region, and comprises complementarity determining regions CDRH1, CDRH2 and CDRH3 of a heavy chain variable region and complementarity determining regions CDRL1, CDRL2 and CDRL3 of a light chain variable region; the monoclonal antibody for the novel coronavirus virus SARS-CoV-2 spike protein RBD region is high in titer and specificity, can be efficiently expressed, can be specifically combined with the spike protein RBD region on the surface of the novel coronavirusvirus SARS-CoV-2, can be used for detecting the novel coronavirus virus SARS-CoV-2, and has a broad application prospect. Certain novel coronavirus virus toxicity can be neutralized and weakened, andthe effect of preventing or / and treating novel coronavirus virus pneumonia is achieved.

Description

technical field [0001] The invention relates to the technical fields of cellular immunology and molecular biology, in particular to a monoclonal antibody directed against the RBD region of the spike protein of the new coronavirus SARS-CoV-2 and its application. Background technique [0002] The new coronavirus SARS-CoV-2, which belongs to the family Coronaviridae, the genus Coronavirus of the subfamily Orthocoronavirinae, is associated with severe acute respiratory syndrome-related coronavirus (SARS-CoV) and Middle East respiratory Syndrome-associated coronavirus (MERS-CoV) close relatives can cause severe pneumonia symptoms. The virus is transmitted through droplets, contact and other means, and patients in the incubation period are contagious. And studies have found that patients with new coronary pneumonia are extremely contagious in the early stages of the disease and when symptoms are mild. [0003] The diameter of the SARS-CoV-2 virus is 75-160 nanometers, and its ge...

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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