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Monoclonal antibody for neutralizing novel coronavirus infection

A coronavirus and antibody technology, applied in antiviral agents, antiviral immunoglobulins, antibodies, etc., can solve problems such as no specific treatment methods

Active Publication Date: 2021-04-09
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For severe cases, there is still no specific treatment method, and serum therapy for patients with new coronary pneumonia is one of the effective treatment methods

Method used

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  • Monoclonal antibody for neutralizing novel coronavirus infection
  • Monoclonal antibody for neutralizing novel coronavirus infection
  • Monoclonal antibody for neutralizing novel coronavirus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Preparation of monoclonal antibody N5 and humanized human-mouse chimeric antibody ch-N5

[0045] 1. Preparation of monoclonal antibody N5

[0046] 1. Discovery of monoclonal antibody N5

[0047] The S protein (100ug / ml) of the SARS-CoV-2 virus was emulsified with an equal volume of Freund's complete adjuvant as an immunogen, and the female Balb / C healthy mice (purchased from military Experimental Animal Center, Academy of Medical Sciences), the injection dose was 0.4 mL of immunogen per mouse. Female Balb-c mice were immunized three times with an interval of 2 weeks between each time. From the second booster immunization, on the 3rd day after each immunization, blood was collected from the orbit of the mouse to measure the antibody titer, and the mouse with the best serum titer was selected, and the spleen of the mouse was dissected to prepare a spleen cell suspension. Cultured in complete DMEM medium for 5 days in an incubator. Hybridoma cells secreting ...

Embodiment 2

[0078] Example 2. Application of monoclonal antibody N5 and humanized human-mouse chimeric antibody ch-N5

[0079] 1. Monoclonal antibody N5 antibody specifically binds to the RBD region of the S protein of the 2019-nCoV novel coronavirus

[0080] The ELISA method is used to detect the binding activity of the monoclonal antibody N5 antibody to the S protein of the new coronavirus, and the operation steps are as follows:

[0081] 1. Coating: Dilute the recombinantly expressed novel coronavirus S-RBD protein (product of Beijing Boaolong Immunology Technology Co., Ltd., BD-VP1488) to 5 μg / ml, and coat 100 μl / well of an ELISA microwell plate, wherein, The formulation of the test solution is: sodium carbonate / sodium bicarbonate buffer solution (0.1M) with pH 9.6. overnight at 4°C.

[0082] 2. Blocking: wash once with washing solution (recipe: phosphate buffered saline solution containing Tween-20 with a volume percentage of 0.5‰, referred to as PBST washing solution), and then us...

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Abstract

The invention discloses a monoclonal antibody for neutralizing novel coronavirus infection. The invention provides the antibody. The amino acid sequence of a light chain variable region of the antibody is the 1-110th site of a sequence 2 in a sequence listing; and the amino acid sequence of a heavy chain variable region of the antibody is the 1-120th site of a sequence 4 in the sequence listing. The monoclonal antibody N5 is identified to have higher binding specificity and affinity to the S protein RBD (receptor binding domain) of the novel coronavirus, and an in vitro neutralization experiment shows that the N5 can effectively neutralize the infection activity of the novel coronavirus SARS-CoV-2. On the basis, the gene sequence of the N5 antibody is measured, then recombinant expression is carried out, and the expressed recombinant antibody also has the neutralizing activity of the SARS-CoV-2.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a monoclonal antibody that neutralizes novel coronavirus infection. Background technique [0002] The novel coronavirus belongs to the Coronaviridae family and has been named SARS-CoV-2. The population is generally susceptible. It has infected more than 70 million people worldwide and killed 300,000 cases. The novel coronavirus is highly contagious, seriously endangers people's health, and can easily cause social panic. The elderly, weak and immunocompromised are more prone to severe illness and high fatality rate. For severe cases, there is still no specific treatment. Serum therapy for convalescent patients with new coronary pneumonia is one of the effective treatment methods. The development of specific therapeutic antibodies has a clear and immediate therapeutic effect in the emergency treatment of infected patients, and has more clinical application value. Contents of the in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13A61K39/42A61P31/14
CPCC07K16/10A61P31/14C07K2317/565C07K2317/56C07K2317/52C07K2317/76C07K2317/24C07K2317/92
Inventor 康晓平李裕昌姜涛李靖张森吴晓燕祝庆余邓永强秦成峰
Owner ACADEMY OF MILITARY MEDICAL SCI
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