High-stability novel coronavirus spike protein, related biological material, application of related biological material, detection test paper and detection kit

A coronavirus, spike protein technology, applied in biomaterial analysis, biochemical equipment and methods, applications, etc., can solve problems such as high false positives, poor specificity, and poor stability, and achieve high accuracy, sensitivity, and accuracy. And the effect of good sensitivity, simple and fast operation

Active Publication Date: 2020-12-15
深圳粒影生物科技有限公司
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AI Technical Summary

Problems solved by technology

However, due to the poor stability of S protein after purification, it cannot be stored for a long time, so the immunological detection reagents currently on the market use N protein or a part of S protein (such as the receptor binding region RBD)
These reagents cannot fully characterize the antigenic properties of the virus

Method used

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  • High-stability novel coronavirus spike protein, related biological material, application of related biological material, detection test paper and detection kit
  • High-stability novel coronavirus spike protein, related biological material, application of related biological material, detection test paper and detection kit
  • High-stability novel coronavirus spike protein, related biological material, application of related biological material, detection test paper and detection kit

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Embodiment Construction

[0040] The present invention constructs a SARS-CoV-2 antigen, which is a full-length S protein in a trimer state;

[0041] The present invention improves the stability of the S protein by introducing cysteine ​​mutations to internally cross-link the S protein in a disulfide bond; the cysteine ​​mutations include: S383C, G413C, D985C and V987C;

[0042] The present invention improves the stability of S protein by introducing a proline mutation; the proline mutation is: K986P;

[0043] The amino acid sequence of the stable full-length S protein carrying mutations in the present invention is shown in SEQ ID NO: 1.

[0044] In order to achieve high-efficiency expression and subsequent purification of the full-length S protein in a stable state, the present invention inserts the following sequences on both sides of the S protein: N-terminal secretion signal peptide, C-terminal polymerization signal peptide and 6xHis affinity tag; the N-terminal secretion The amino acid sequence of...

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Abstract

The invention discloses a high-stability novel coronavirus spike protein, a related biological material, application of the related biological material, detection test paper and a detection kit, relates to the field of biotechnology and biomedicine. The bottleneck of the industry is broken through through internal crosslinking modification on the optimal antigen spike protein, namely S protein, onthe surface of novel coronavirus, The high-stability full-length S protein trimer is obtained through expression and purification in mammals, all antigen epitopes of the new coronavirus surface spikeprotein are reserved to the maximum extent, the stable S protein serves as an antigen to capture a new coronavirus antibody in a patient sample, and a novel coronavirus antibody instant detection kitwith high accuracy, high sensitivity and high stability is produced.

Description

technical field [0001] The present invention relates to the fields of biotechnology and biomedicine, in particular to a highly stable novel coronavirus spike protein, related biological materials and applications thereof, detection test strips and detection kits, and to a stable S protein novel coronavirus based on internal cross-linking. Quantitative real-time detection kits for viral antibodies, specifically applying the best antigen stable full-length S protein on the surface of the new coronavirus, colloidal gold and nano-magnetic particle technology, to realize the anti-new coronavirus S protein IgM antibody and (or) in the blood Test strips and kits for point-of-care detection of IgG antibodies. Background technique [0002] Novel coronavirus pneumonia (COVID-19) is an infectious disease caused by infection with a new type of coronavirus (SARS-CoV-2). [0003] The novel coronavirus is highly transmissible, has a long incubation period, and is generally susceptible to ...

Claims

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Application Information

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IPC IPC(8): C07K14/165C12N15/50G01N33/569G01N33/558G01N33/543
CPCC07K14/005G01N33/56983G01N33/558G01N33/54313C12N2770/20022G01N2333/165
Inventor 张影熊晓犁李宁宁张秦鲁徐涤非范恒新陈明徐朋奇何俊周光前
Owner 深圳粒影生物科技有限公司
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