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Monoclonal antibody for non-RBD region of new coronavirus spinous process protein and application of monoclonal antibody

A monoclonal antibody, spike protein technology, applied in the field of bioengineering, can solve problems such as insignificant effects, toxic and side effects, and achieve the effect of simple and efficient screening, strong specificity, prevention or/or treatment of new coronavirus pneumonia

Active Publication Date: 2020-12-01
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 for the treatment of new coronary pneumonia, and the effect of conventional antibiotic drug treatment is not obvious, and some of them have serious side effects

Method used

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  • Monoclonal antibody for non-RBD region of new coronavirus spinous process protein and application of monoclonal antibody
  • Monoclonal antibody for non-RBD region of new coronavirus spinous process protein and application of monoclonal antibody
  • Monoclonal antibody for non-RBD region of new coronavirus spinous process protein and application of monoclonal antibody

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Experimental program
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Effect test

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 109 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 bacteriophage containing 100μg / ml ampicillin, 50μg / 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 mi...

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Abstract

The invention discloses a monoclonal antibody for a non-RBD region of a new coronavirus spinous process protein and application of the monoclonal antibody. The monoclonal antibody specifically binds to the RBD region of the new coronavirus spinous process glycoprotein, 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 non-RBD region of a new coronavirus spinous process protein is high in titer and specificity, can be efficiently expressed, can be specifically combined with a region outside a spinous process protein RBD region on the surface of new coronavirus SARS-CoV-2, is used for qualitative or quantitative detection of the new coronavirus, can neutralize and weaken certain toxicity of the new coronavirus, and plays a role in preventing or / and treating the new coronavirus 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 envelope structure of the new coronavirus contains three proteins: spike protein (Spike protein, S protein), membrane (Membrane protein, M protein) and envelope protein (Envelope protein, E protein). M protein is involved in the formation of the virus envelope It is related to the budding process; HE protein constitutes the short protrusion of the envelope, which may be related to the early adsorption of coronaviruses. The HE protein of some coronaviruses can cause erythrocyte agglutination and erythrocyte adsorption; It plays a key role in guiding the fusion of the viral envelope with the cell membrane. In many mammals, the S p...

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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