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Construction method and application of coronavirus antigen on the basis of mosaic strategy

A coronavirus and human coronavirus technology, applied in the direction of viral antigen components, viruses, viral peptides, etc., can solve the problems of ignoring the importance of T-cell immune response and no vaccine products.

Inactive Publication Date: 2020-08-14
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, considering factors such as safety and efficacy, there is no vaccine product against HCoV
[0003] The importance of the T cell immune response has often been overlooked in the development of coronavirus vaccines in the past

Method used

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  • Construction method and application of coronavirus antigen on the basis of mosaic strategy
  • Construction method and application of coronavirus antigen on the basis of mosaic strategy
  • Construction method and application of coronavirus antigen on the basis of mosaic strategy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Analysis of Conserved CTL Epitopes on HCoVs Structural Proteins

[0081] In order to study the conserved T cell epitopes that exist on all known HCoVs, this embodiment first downloads a total of 3132 coronavirus genome sequences of all regions, all times, and all hosts from NCBI, VIPR and GISAID, including 928 HCoVs and 2204 other CoVs, and focused on the analysis of four structural proteins encoded by the HCoVs genome: S protein (spike-surface glycoprotein), E protein (small envelope protein), M protein (matrix protein) and N protein (nucleocapsid protein);

[0082] After using computer algorithms to remove repetitive sequences and sequences with poor sequencing quality, 534 S protein sequences, 485 M protein sequences, 410 N protein sequences and 478 E protein sequences were obtained, and the NetMHCpan-4.0 tool was used to predict the sequence of these proteins Potential cytotoxic T lymphocyte (CTL) epitopes present;

[0083] According to the phenotype freq...

Embodiment 2

[0086] Example 2 Design of Mosaic Antigen and Analysis of CTL Epitopes

[0087] In order to obtain artificial antigens that cover conserved CTL epitopes to the greatest extent, this example uses the Mosaic VaccineDesigner program to further analyze the S, M, N, and E protein sequences, and removes rare, low-frequency occurrences in natural epitopes (the number of occurrences is less than 3) After the epitope was identified, a series of mosaic antigens assembled from short peptides (composed of 9 amino acids) were obtained: S protein mosaic cocktail (SEQ ID NO: 1-4), M protein mosaic cocktail (SEQ ID NO: 5- 8), N protein mosaic cocktail (SEQ ID NO: 9-12) and E protein mosaic cocktail (SEQ ID NO: 13-16).

[0088] Analysis of the CTL epitopes on the four mosaic antigens revealed that most of the CTL epitopes on the natural S, M, N, E proteins were present on the mosaic antigens. Such as figure 1 As shown, 9 amino acids of 90% CTL epitopes on the mosaic antigen completely match ...

Embodiment 3

[0092] Phylogenetic Analysis of Example 3 Mosaic Antigens

[0093] This example explores the relationship between mosaic antigens and natural proteins. Such as image 3 As shown, 7 HCoVs form 4 clades, among which, A is the clade of HCoV-229E and HCoV-NL63, B is the clade of SARS-CoV and SARS-CoV-2, and C is the clade of HCoV-HKU1 and HCoV- The clade of OC43, D is the clade of MERS-CoV; the four mosaic antigens (stars) exist in each clade, indicating that the designed mosaic antigens widely cover all known HCoVs.

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Abstract

The invention provides a construction method and application of a coronavirus antigen on the basis of a mosaic strategy. The method comprises the following steps of: (1) analyzing the natural structural protein of a human coronavirus, and obtaining cytotoxic T lymphocyte epitope which is in the presence on the natural structural protein; and (2) adopting the mosaic strategy to analyze the structural protein of the human coronavirus, and after rare epitope with low occurrence frequency in the natural structural protein is removed, obtaining the coronavirus antigen assembled by short peptides. According to the construction method, on the basis of an artificial intelligence algorithm, all known CoV (coronavirus) genomic sequences are subjected to big data analysis, and a novel antigen sequence is designed on the basis of the mosaic strategy. A novel antigen covers most CTL (cytotoxic T lymphocyte) epitope of natural S, M, N and E proteins on all seven known HCoVs (human coronavirus), hasa structural similarity with a natural protein, and has a potential application prospect and value on an aspect of preparing a coronavirus universal vaccine.

Description

technical field [0001] The invention belongs to the field of biomedicine technology, and relates to a method for constructing a coronavirus antigen based on a mosaic strategy and its application, in particular to a method for constructing a coronavirus antigen based on a mosaic strategy, and a coronavirus covering a large number of conserved T cell epitopes thus constructed Antigen and its application in the preparation of a coronavirus universal vaccine. Background technique [0002] At present, the methods for researching human coronavirus (HCoV) vaccines mainly include inactivated vaccines, live attenuated vaccines, subunit vaccines, DNA vaccines, mRNA vaccines, recombinant vector vaccines, and combination vaccines. However, considering factors such as safety and efficacy, there is no vaccine product against HCoV. [0003] In the development of coronavirus vaccines in the past, the importance of T cell immune response was often ignored. T cell immune response is a k...

Claims

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Application Information

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IPC IPC(8): C07K14/165C07K19/00A61K39/215A61P31/14A61P11/00
CPCC07K14/005A61K39/12A61P31/14A61P11/00C12N2770/20022C12N2770/20034C07K2319/40A61K2039/53
Inventor 孙彩军杜向军舒跃龙李敏超曾金锋
Owner SUN YAT SEN UNIV
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