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Antibody aiming at novel coronavirus NP protein and detection application thereof

A coronavirus, a new type of technology, applied in the fields of cellular immunology and molecular biology, can solve the problems of false positives and false negatives, affecting the implementation of prevention and control measures, and being prone to missed diagnosis.

Active Publication Date: 2020-08-21
JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although nucleic acid diagnosis is fast, it is greatly affected by the quality of sampling, and there are false positives and false negatives, which affect the implementation of prevention and control measures
Some asymptomatic infected persons are also negative in the nucleic acid test in the late stage of the disease, and it is easy to miss the diagnosis based on the nucleic acid test alone

Method used

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  • Antibody aiming at novel coronavirus NP protein and detection application thereof
  • Antibody aiming at novel coronavirus NP protein and detection application thereof
  • Antibody aiming at novel coronavirus NP protein and detection application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Antibody Screening

[0073] 1. Expression of recombinant SARS-CoV2 nucleoprotein (NP)

[0074] 1.1 Main reagents

[0075] The SARS-CoV2 NP gene sequence (GenBank serial number: MT066176.1) and related primers were synthesized and sequenced by General Biosystems (Anhui) Co., Ltd.; E.coli DH5α, BL21(DE3) competent cells were purchased from General Biosystems (Anhui) Co., Ltd.; BamHI and NotI endonucleases were purchased from New England Biolabs (NEB); EX Taq enzyme was purchased from TaKaRa Company; HRP-labeled anti-human Fc antibody was purchased from Sigma Company; other chemical reagents were domestic analytical grade Reagents; Sera from patients infected with 2019-nCoV were collected and preserved by our center, all of which were Jiangsu cases.

[0076] 1.2 Prokaryotic expression plasmid construction

[0077] The primers for prokaryotic expression of NP gene were designed, the upstream primer had a BamHI restriction site, and the downstream primer had a N...

Embodiment 2

[0127] Example 2 Expression of whole antibody and related functional verification

[0128] From the obtained 159 antibodies, 16 antibodies were finally selected for the expression of the whole antibody and related functional verification, and these 16 antibodies were named JS01-JS16.

[0129] Among them, the JS13 antibody sequence is as follows:

[0130] The amino acid sequence of the heavy chain variable region CDR1 is shown in SEQ ID NO:1;

[0131] The amino acid sequence of the heavy chain variable region CDR2 is shown in SEQ ID NO:2;

[0132] The amino acid sequence of the heavy chain variable region CDR3 is shown in SEQ ID NO: 3;

[0133] The amino acid sequence of the light chain variable region CDR1 is shown in SEQ ID NO:5;

[0134] The amino acid sequence of the light chain variable region CDR2 is shown in SEQ ID NO: 6;

[0135] The amino acid sequence of the light chain variable region CDR3 is shown in SEQ ID NO:7;

[0136] The amino acid sequence of the heavy ch...

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Abstract

The invention discloses an antibody aiming at a novel coronavirus NP protein and a detection application thereof. The antibody comprises an antigen complementation determining region CDR1, an antigencomplementation determining region CDR2 and an antigen complementation determining region CDR3 of an antibody light chain variable region, wherein the antigen complementation determining region CDR1,the antigen complementation determining region CDR2 and the antigen complementation determining region CDR3 are respectively amino acid sequences shown in SEQ ID NO: 5, SEQ ID NO: 6 and SEQ ID NO: 7;the antigen complementation determining regions CDR1, CDR2 and CDR3 of the antibody heavy chain variable region are respectively amino acid sequences shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO:3. The invention also discloses a preparation process of the antibody and amino acid sequences of a heavy chain variable region and a light chain variable region of the antibody.

Description

technical field [0001] The invention belongs to the fields of cellular immunology and molecular biology, and relates to an antibody against a novel coronavirus NP protein and its detection application. Background technique [0002] The International Committee on Taxonomy of Viruses named the new coronavirus as SARS-CoV-2, and the World Health Organization named the pneumonia caused by this virus as COVID-19. The virus is highly contagious and spreads widely. The virus can quickly adapt to the human environment, and has the ability to spread during the incubation period after infection. At the same time, some asymptomatic infections have been reported, and viral nucleic acids have even been detected in various animals. These factors make the prevention and control of the virus very complicated, and there are currently no effective therapeutic drugs and vaccines on the market. [0003] SARS-CoV-2 belongs to the genus Coronaviridae and is a single-stranded positive-sense RNA ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/569G01N33/577
CPCC07K16/005C07K16/10C07K2317/565C07K2317/92G01N33/56983G01N33/577G01N2333/165
Inventor 张黎郑滨洋高行素郭喜玲朱凤才王祥喜朱玲孙瑶李靖欣
Owner JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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