Method for preparing monoclonal antibody of spike protein of novel coronavirus

A monoclonal antibody and spike protein technology, applied in antiviral immunoglobulin, virus/phage, biochemical equipment and methods, etc., can solve the problem of no effective drugs, reduce the cure rate of SARS-CoV-2 virus, etc. question

Inactive Publication Date: 2021-01-05
通用生物(安徽)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The SARS-CoV-2 virus that appears at this stage has a very strong infectious ability, and there is no effective drug that can eliminate the SARS-CoV-2 virus, so that patients can only be cured by their own resistance, which greatly reduces the SARS-CoV- 2 The cure rate of the virus

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  • Method for preparing monoclonal antibody of spike protein of novel coronavirus
  • Method for preparing monoclonal antibody of spike protein of novel coronavirus
  • Method for preparing monoclonal antibody of spike protein of novel coronavirus

Examples

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Embodiment 1

[0024]A preparation method of a novel coronavirus spike protein monoclonal antibody, the specific steps are as follows:

[0025]1 Antigen preparation: Complete gene synthesis of Skipe protein RBD region sequence, subcloned into eukaryotic expression vector, and add 8*His tag to C-terminal of expression vector to express purified spike protein receptor binding domain SP-RBD -His;

[0026]2 Animal immunization: Balb / c mice were immunized with spike protein receptor binding domain SP-RBD-His, each Balb / c mouse was immunized with 50 μg each time, immunized 3 times, each time interval was two weeks;

[0027]3 Cell fusion: Take the mouse spleen 1 week after three immunizations, put the mouse spleen into a petri dish containing 5mL DMEM solution, crush the spleen, filter with nylon mesh to obtain a single cell suspension, and suspend the single cell The liquid and logarithmic phase mouse myeloma cell SP2 / 0 cell liquid were mixed at a volume ratio of 1:1, and the cell fusion was performed with a cel...

Embodiment 2

[0032]A preparation method of a novel coronavirus spike protein monoclonal antibody, the specific steps are as follows:

[0033]1 Antigen preparation: Complete gene synthesis of Skipe protein RBD region sequence, subcloned into eukaryotic expression vector, and add 8*His tag to C-terminal of expression vector to express purified spike protein receptor binding domain SP-RBD -His;

[0034]2 Animal immunization: Balb / c mice were immunized with spike protein receptor binding domain SP-RBD-His, each Balb / c mouse was immunized with 50 μg each time, immunized 3 times, each time interval was two weeks;

[0035]3 Cell fusion: Take the mouse spleen 1 week after three immunizations, put the mouse spleen into a petri dish containing 5mL DMEM solution, crush the spleen, filter with nylon mesh to obtain a single cell suspension, and suspend the single cell The liquid and logarithmic phase mouse myeloma cell SP2 / 0 cell liquid were mixed at a volume ratio of 1:1, and the cell fusion was performed with a cel...

Embodiment 3

[0040]A preparation method of a novel coronavirus spike protein monoclonal antibody, the specific steps are as follows:

[0041]1 Antigen preparation: Complete gene synthesis of Skipe protein RBD region sequence, subcloned into eukaryotic expression vector, and add 8*His tag to C-terminal of expression vector to express purified spike protein receptor binding domain SP-RBD -His;

[0042]2 Animal immunization: Balb / c mice were immunized with spike protein receptor binding domain SP-RBD-His, each Balb / c mouse was immunized with 50 μg each time, immunized 3 times, each time interval was two weeks;

[0043]3 Cell fusion: Take the mouse spleen 1 week after three immunizations, put the mouse spleen into a petri dish containing 5mL DMEM solution, crush the spleen, filter with nylon mesh to obtain a single cell suspension, and suspend the single cell The liquid and logarithmic phase mouse myeloma cell SP2 / 0 cell liquid were mixed at a volume ratio of 1:1, and the cell fusion was performed with a cel...

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Abstract

The invention discloses a method for preparing a monoclonal antibody of a spike protein of novel coronavirus. The method comprises the following steps: antigen preparing, animal immunizing, cell fusing, screening detecting, stable cell line establishing and monoclonal antibody producing. The spike protein is a superficial membrane protein of SARS-CoV-2 and contains two subunits S1 and S2, the S1 mainly comprises one receptor binding domain and is responsible for identifying a cell surface receptor, and the S2 comprises fundamental elements required for membrane fusion. The S protein reacts with an antibody and T cells in induction and plays a crucial role in protective immunization, so that the monoclonal antibody of the spike protein can be excellently bound to the spike protein to form acomplex, the complex can be digested or degraded, then, the SARS-CoV-2 virus can be deactivated, and the effect of killing the SARS-CoV-2 virus is achieved.

Description

Technical field[0001]The invention belongs to the technical field of monoclonal antibody preparation, and specifically relates to a preparation method of a novel coronavirus spike protein monoclonal antibody.Background technique[0002]Since December 2019, many cases of pneumonia of unknown cause have been discovered one after another, which has now been confirmed as an acute respiratory infection caused by a new type of coronavirus infection. Based on the current epidemiological survey, the incubation period is generally 3-7 days, and the longest does not exceed 14 days. The clinical manifestations of this epidemic are mainly fever, fatigue, and dry cough. A small number of patients are accompanied by nasal congestion, runny nose, diarrhea and other symptoms. Some patients only showed low fever, mild fatigue, etc., without pneumonia, and recovered after 1 week. The World Health Organization named the pathogen that caused this viral pneumonia case as the 2019 new coronavirus, which is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K19/00
CPCC07K14/005C07K16/10C07K2317/14C07K2319/21C12N2770/20022
Inventor 雍金贵张磊缪连军
Owner 通用生物(安徽)股份有限公司
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