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B cell linear epitope of novel coronavirus S protein, antibody, identification method and application

A coronavirus and antigenic epitope technology, applied in the field of biology, can solve the problems of low accuracy and limited research data on SARS-CoV-2 antigenic epitopes

Inactive Publication Date: 2021-01-08
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Recent reports have identified some of the SARS-CoV-2 S protein B cell linear epitopes, providing information on the distribution of some of the SARS-CoV-2 B cell linear epitopes. The information is still very limited, and most reports are only based on software predictions, with low accuracy

Method used

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  • B cell linear epitope of novel coronavirus S protein, antibody, identification method and application
  • B cell linear epitope of novel coronavirus S protein, antibody, identification method and application
  • B cell linear epitope of novel coronavirus S protein, antibody, identification method and application

Examples

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Embodiment 1

[0037] Example 1: Identification method of B cell linear epitope of novel coronavirus S protein

[0038] 1. Establishment of peptide library

[0039] According to the amino acid sequence of SARS-CoV-2 S protein (access number: YP_009724390.1), the synthetic polypeptide was designed. According to the sequence of amino acids, peptides with a length of 15 amino acids were synthesized, and adjacent peptides overlapped by 5 amino acids. A total of 127 peptides (P1-P127) were obtained, covering the full length of S protein. Peptides were divided into 13 groups (G1-G13) according to the sequence of amino acid positions, and each group contained 10 or 7 consecutive peptides (Table 1).

[0040] Table 1 SARS-CoV-2 overlapping peptide library sequence (peptide library)

[0041]

[0042]

[0043]

[0044]

[0045]2. Collection of serum from COVID-19 patients during recovery period

[0046] From March 5, 2020 to May 12, 2020, serum was collected from 165 discharged patients...

Embodiment 2

[0056] Example 2: Detection of novel coronavirus antibody with B cell linear epitope, embodiment of detection of novel coronavirus antibody comprising B cell linear epitope P104

[0057] Four B cell linear epitopes (S21P2, S14P5, P82, and P104) were coated on 96-well plates, and 165 convalescent sera from COVID-19 patients were tested by indirect ELISA one by one. Each sample was measured in duplicate. The cutoff value was calculated using healthy human serum as a negative control, and then the S / CO value (sample average OD450nm-630nm value / cutoff value) was calculated.

[0058] Among them, the main operation of indirect ELISA is as follows: ELISA 96-well plate (Corning, #42592) is used for antigen coating, and 100 μL of peptides (each peptide 2.5 μg / mL) or 0.2 μg / mL RBD (CUSABIO, #CSB- MP3324GMY1b1), overnight at 4°C. The plate was washed 6 times with 300 μL of washing buffer (PBS containing 0.05% Twenn-20). Add 300 μL of blocking solution (PBS containing 0.1% Tween-20 and...

Embodiment 3

[0066] Embodiment 3: novel coronavirus vaccine

[0067] The antigenic epitope P104 is used in the preparation of the new coronavirus vaccine, that is, the antigenic epitope P104 polypeptide is used to immunize humans or animals, and the vaccinated humans or animals will produce corresponding antibodies, and the production of antibodies is the main purpose of vaccination. For a specific implementation manner, reference may be made to the prior art, and details are not repeated here.

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Abstract

The invention is applicable to the technical field of biology, and provides a B cell linear epitope of a novel coronavirus S protein, an antibody, an identification method and an application, and theamino acid sequence of the B cell linear epitope of the S protein is shown as SEQ ID NO.104 or SEQ ID NO.82. The invention provides a brand-new B cell linear epitope of the novel coronavirus S protein, which can be used for detecting a specific antibody in blood of a COVID-19 patient, and can also be used for being inoculated into healthy people to induce generation of the specific antibody, so that an important antigen target is provided for design of an SARS-CoV-2 vaccine, research and development of the monoclonal antibody and research and development of an antibody detection kit.

Description

technical field [0001] The invention belongs to the technical field of biology, and in particular relates to a B cell linear antigen epitope, identification method and application of novel coronavirus S protein. Background technique [0002] The novel coronavirus disease 2019 (COVID-19) that broke out in December 2019 was caused by SARS-CoV-2 (severe acute respiratory syndrome corona virus 2). COVID-19 has become a global pandemic, and the epidemic has spread to more than 200 countries and the situation is severe. [0003] The entire genome of SARS-CoV-2 is about 29.8kb nucleotides in length, encoding four main structural proteins: Spike protein S (Spike protein, S protein), membrane protein M (Membrane protein), envelope protein E (Envelope protein ) and nucleocapsid protein N (Nucleocapsid). Among them, after SARS-CoV-2 mediates the binding of the virus to the cell receptor through the receptor binding domain (RBD) on the S protein, the S1 and S2 subunits are cut and dis...

Claims

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Application Information

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IPC IPC(8): C07K14/165C07K16/10G01N33/68G01N33/569A61K39/215A61P31/14
CPCC07K14/005C07K16/10G01N33/6854G01N33/56983A61K39/12A61P31/14C12N2770/20022C12N2770/20034C07K2317/34G01N2333/165G01N2469/20
Inventor 王晓辉汪川张云雯杨峥嵘冯铁建扈庆华何建凡
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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