Novel coronavirus antibody and ELISA detection kit of novel coronavirus antibody

A coronavirus, a new type of technology, applied in the field of cellular immunology and molecular biology, can solve the problems of large impact, prone to missed diagnosis, false positive and false negative, etc.

Active Publication Date: 2020-08-11
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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  • Novel coronavirus antibody and ELISA detection kit of novel coronavirus antibody
  • Novel coronavirus antibody and ELISA detection kit of novel coronavirus antibody
  • Novel coronavirus antibody and ELISA detection kit of novel coronavirus antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Antibody Screening

[0079] 1. Expression of recombinant SARS-CoV2 nucleoprotein (NP)

[0080] 1.1 Main reagents

[0081] 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); EXTaq enzyme was purchased from TaKaRa; HRP-labeled anti-human Fc antibody was purchased from Sigma; other chemical reagents were domestic analytical reagents ; Sera from patients infected with 2019-nCoV were collected and preserved by our center, all of which were Jiangsu cases.

[0082] 1.2 Prokaryotic expression plasmid construction

[0083] The primers for prokaryotic expression of NP gene were designed, the upstream primer had a BamHI restriction site, and the downstream primer had a NotⅠ restriction site. ...

Embodiment 2

[0134] Example 2 Expression of whole antibody and related functional verification

[0135] 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.

[0136] Among them, the JS12 antibody sequence is as follows:

[0137] The amino acid sequence of the heavy chain variable region CDR1 is shown in SEQ ID NO.1;

[0138] The amino acid sequence of the heavy chain variable region CDR2 is shown in SEQ ID NO.2;

[0139] The amino acid sequence of the heavy chain variable region CDR3 is shown in SEQ ID NO.3;

[0140] The amino acid sequence of the light chain variable region CDR1 is shown in SEQ ID NO.5;

[0141] The amino acid sequence of the light chain variable region CDR2 is shown in SEQ ID NO.6;

[0142] The amino acid sequence of the light chain variable region CDR3 is shown in SEQ ID NO.7;

[0143] The amino acid sequence of the heavy chai...

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Abstract

The invention discloses a novel coronavirus antibody and an ELISA detection kit of the novel coronavirus antibody. The antibody comprises CDR1 as shown in SEQ ID NO.1, CDR2 as shown in SEQ ID NO.2, CDR3 as shown in SEQ ID NO.3 and CDR4 as shown in SEQ ID NO.4, CDR1 shown in SEQ ID NO.5, CDR2 shown in SEQ ID NO.6, and CDR3 shown in SEQ ID NO.7. The antibody provided by the invention can be used fornovel coronavirus detection or novel coronavirus infection diagnosis.

Description

technical field [0001] The invention belongs to the fields of cellular immunology and molecular biology, and relates to a novel coronavirus antibody and an ELISA detection kit for the novel coronavirus antibody. 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 treatments and vaccines on the market. [0003] SARS-CoV-2 belongs to the genus Coronaviridae and is a single-stranded positive-sense R...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/569G01N33/543
CPCC07K16/10C07K2317/56C07K2317/565G01N33/543G01N33/56983G01N2333/165
Inventor 张黎高行素郑滨洋孟繁岳王文娟郭喜玲梁祁朱凤才
Owner JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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