Monoclonal antibody capable of aiming at SARS-CoV-2 spike protein non RBD (Receptor Binding Domain), and application of monoclonal antibody

A monoclonal antibody and spike protein technology, which is applied in the fields of molecular biology and cellular immunology, can solve problems such as the listing of therapeutic drugs and vaccines, and the specificity of new coronaviruses, and achieve weakened virus toxicity, strong specificity, and high potency Effect

Active Publication Date: 2020-11-20
潍坊市第二人民医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, there are many treatment options for new coronary pneumonia, but there are no specific therapeutic drugs and vaccines for new coronavirus on the market. Since antibody drugs have played an important role in the treatment of infectious diseases, autoimmunity and other diseases, it also gives us research Antibodies against viruses provide theoretical support, so it is of great significance to develop monoclonal antibodies against the non-RBD region of the SARS-CoV-2 spike protein of the new coronavirus

Method used

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  • Monoclonal antibody capable of aiming at SARS-CoV-2 spike protein non RBD (Receptor Binding Domain), and application of monoclonal antibody
  • Monoclonal antibody capable of aiming at SARS-CoV-2 spike protein non RBD (Receptor Binding Domain), and application of monoclonal antibody
  • Monoclonal antibody capable of aiming at SARS-CoV-2 spike protein non RBD (Receptor Binding Domain), 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 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, discard the supernatant, ad...

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Abstract

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

Description

technical field [0001] The invention relates to the technical fields of cellular immunology and molecular biology, in particular to a monoclonal antibody against the non-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 Coronaviridae family (Coronaviridae), the betacoronavirus genus (Coronavirus) of the Orthocoronavirinae subfamily (Orthocoronavirinae), the coronavirus associated with severe acute respiratory syndrome (SARS-CoV) and the Middle East Respiratory Syndrome-associated coronavirus (MERS-CoV) close relatives can cause severe pneumonia symptoms. Common clinical symptoms after human infection with the new coronavirus mainly include fever, fatigue, dry cough as the main respiratory symptom, and gradually dyspnea, acute respiratory distress in severe cases, metabolic acidosis that is difficult to correct, and coagulation dysfunction, etc. [0003] Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10G01N33/577G01N33/569A61K39/42A61P31/14A61P11/00
CPCA61K2039/505A61P11/00A61P31/14C07K16/10C07K2317/35C07K2317/56C07K2317/565C07K2317/76G01N33/56983G01N33/577G01N2333/165G01N2469/10
Inventor 董金华陈丽梅李海梅
Owner 潍坊市第二人民医院
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