SARS-CoV-2 lymphocyte antigen epitope peptides and application thereof

A sars-cov-2 and epitope peptide technology, applied in the field of medical immunology and infectious diseases, can solve the problems of restricting the development of vaccines and therapeutic preparations, restricting the study of immune pathology and immune protection mechanism

Inactive Publication Date: 2021-06-15
SOUTHEAST UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently, there are still few SARS-CoV-2 antigenic epitope peptides that can stimulate T cell responses, which limits the monitoring and analysis of specific T cell immune function in patients with SARS-CoV-2 infection, and also limits The researc

Method used

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  • SARS-CoV-2 lymphocyte antigen epitope peptides and application thereof
  • SARS-CoV-2 lymphocyte antigen epitope peptides and application thereof
  • SARS-CoV-2 lymphocyte antigen epitope peptides and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1: In some examples of the present invention, a method for predicting and identifying the amino acid sequence of a SARS-CoV-2 protein T cell epitope peptide is provided, and the specific steps are as follows:

[0038] 1. Virtual prediction of T cell epitope peptides restricted by HLA-A molecules from 5 SARS-CoV-2 proteins

[0039] Select 5 kinds of SARS-CoV-2 proteins, for example, E protein (envelope protein), M protein (membrane protein), N protein (nucleocapsid protein), S protein (spike protein) and RdRp protein (RNA-dependent RNA polymerase ). Then use SYFPEITHI, EPIJEN, ConvMHC, IEDB (including ANN, Consensus, NetMHCpan, SMM, SMMPMBEC) and other epitope peptide prediction tools to predict HLA-A0201, A1101, A2402, A3101, A0206, A0207, A3303, A3001, A0203, A1102, A0301, A0101, A2601 and other molecularly restricted T cell epitope peptides for each of the above proteins were virtually predicted.

[0040] The two ends of the antigen-binding groove of the ...

Embodiment 2

[0099] Example 2: In some examples of the present invention, part of the epitope peptides can be synthesized from the above 164 amino acid sequences, namely the T cell epitope peptide sequences shown in SEQ ID NO: 1-164, to prepare a mixed polypeptide vaccine. As an example, the preparation steps are as follows:

[0100] 1. Prepare the mixed peptide pool and adjuvant first

[0101] 1) Using the T cell epitope peptide sequences of SEQ ID NO: 1-164, synthesize 31 kinds of epitope peptides that can be presented by HLA-A2 molecules. Peptides can be synthesized by Suzhou Qiangyao Biotechnology Co., Ltd., with a purity of >95%, 4mg / peptide, 1mg per tube, and stored at -800°C for later use;

[0102] 2) Dissolving peptides: Take 1 tube (1 mg) of each peptide and store it at room temperature for pre-warming. Dissolve it with 20 μL DMSO, then add 180 μL PBS to make the peptide storage concentration 5 μg / μL, store at -80°C for later use.

[0103] 3) In this example, for example, the fo...

Embodiment 3

[0147] Example 3: In some examples of the present invention, epitope peptides were synthesized using the T cell epitope peptide sequences described in SEQ ID NO: 1-164, 10 peptide libraries were formed, and ELISPOT method was established to detect healthy blood donors The number of specific T cells that cross-react with the T cell epitope peptide of the SARS-CoV-2 protein in the population verifies the feasibility of using the above-mentioned expressed peptides to prepare reagents to detect the immune function of SARS-CoV-2 specific T cells. The specific steps can be as follows:

[0148] 1) Divide the above 164 epitope peptides into 10 peptide libraries according to the type of source protein, in which the epitope peptides of each protein are divided into 2 peptide libraries (23 epitope peptides for E protein, 36 epitope peptides for M protein , 25 kinds of epitope peptides of N protein, 41 kinds of epitope peptides of S protein, 39 kinds of epitope peptides of RdRp), each pep...

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Abstract

The invention discloses lymphocyte antigen epitope peptides and application thereof, belongs to the fields of medical immunology and infectious diseases, and particularly relates to 164 thymus dependent lymphocyte antigen epitope peptides of five proteins of SARS-CoV-2 and application thereof. The antigen epitope peptides can be presented by HLA-A molecules to stimulate activation, proliferation and differentiation of SARS-CoV-2 specific thymus dependent lymphocytes, so that the immune effect of resisting SARS-CoV-2 infection is exerted. The antigen peptides can be used for preparing mixed polypeptide vaccines of SARS-CoV-2, recombinant protein vaccines connected with a plurality of epitope peptides in series and DNA and RNA vaccines, can be used for preparing a detection kit for detecting SARS-CoV-2 specific thymus dependent lymphocytes, can also be used for preparing effector thymus dependent lymphocytes or drugs for treating SARS-CoV-2 infection, and have a potential application value in the prevention, treatment and diagnosis of SARS-CoV-2 infection and COVID-19.

Description

technical field [0001] The invention relates to the fields of medical immunology and infectious diseases, in particular to a SARS-CoV-2 lymphocyte antigen epitope peptide and its application. Background technique [0002] SARS-CoV-2 is an enveloped, single-stranded positive-strand RNA betacoronavirus of the Coronaviridae family, with a genome length of 30,000 bases. Including 14 open reading frames (ORFs), Orf1ab at the 5' end encodes 16 non-structural proteins (Nsp1-16), of which Nsp7, Nsp8 and Nsp12 constitute the RNA-dependent RNA polymerase (RNA- dependentRNA polymerase, RdRp), the full length is 932 amino acids. The 13 Orfs at the 3' end encode 4 structural proteins and 9 putative cofactors, respectively. Among them, the four viral structural proteins are: S protein (spike protein), with a total length of 1273 amino acids; E protein (envelope protein), with a total length of 75 amino acids; M protein (membrane protein), with a total length of 222 amino acids; N protei...

Claims

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Application Information

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IPC IPC(8): C07K14/165C12N5/0783A61K39/215A61P31/14C12Q1/02
CPCC07K14/005C12N5/0636A61K39/12A61P31/14G01N33/505C12N2770/20022C12N2770/20034C12N2501/999A61K2039/5158G01N2333/165
Inventor 沈传来金萧萧丁艳孙世慧赵光宇何玉先
Owner SOUTHEAST UNIV
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