Novel coronavirus antigen epitope and application thereof

A technology of antigenic epitopes and coronaviruses, applied in the direction of viral antigen components, viruses, viral peptides, etc., can solve problems such as affecting the immune response of viruses and changing the immunogenicity of viruses

Active Publication Date: 2020-10-30
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Since mutations in viral proteins may alter the immunogenicity of the virus and potentiall

Method used

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  • Novel coronavirus antigen epitope and application thereof
  • Novel coronavirus antigen epitope and application thereof
  • Novel coronavirus antigen epitope and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Antigen epitope calculation

[0075] This embodiment is based on the whole genome nucleic acid sequence of SARS-CoV-2 (GeneBank: MN988669.1), and the original pdb files of S protein, M protein, E protein and N protein are established by the method of homology modeling; in the original pdb file Based on adding water and adjusting the pH of Cl - and Na + , run the molecular dynamics simulation program to obtain the pdb file under the state of human body temperature (310K); based on the pdb file under the state of human body temperature, the molecular dynamics software Gromacs is used to calculate the all-atom structure of the protein, and the following is obtained: Figure 1A , Figure 1B , Figure 1C with Figure 1D The molecular orbital energy level and spatial structure information of the S protein, M protein, E protein and N protein shown;

[0076] Based on the spatial structure information of the structural models of S protein, M protein, E protein and ...

Embodiment 2

[0077] Homology Analysis of Example 2 Epitope

[0078] The amino acid sequences of the S protein, M protein, E protein, and N protein of SARS-CoV-2 were compared with the corresponding amino acid sequences of the S protein, M protein, E protein, and N protein of SARS-CoV and RaTG13 using the ClustalW program to gradually compare To compare the methods, the steps are as follows:

[0079] (1) Input the protein sequence, set the Gap Open Penalty parameter to 10, the Gap Extension Penalty parameter to 0.05, and the scoring matrix to BLOSUM, and compare the three sequences pairwise to construct a relationship matrix;

[0080] (2) Generate a phylogenetic tree based on the distance matrix, start from the two closest sequences, gradually introduce adjacent sequences to construct an alignment, and obtain a protein homology analysis diagram;

[0081] (3) Mark the selected antigenic epitope on the protein homology analysis map, and analyze the conservation of the antigenic epitope among...

Embodiment 3

[0086] Example 3 Utilizes HBc-S to prepare a VLP vaccine displaying antigenic epitopes

[0087] Synthesize the epitope-SpyTag, mix it with the HBc VLP expressing SpyCatcher at a molar ratio of 3:1, add 3 times the volume of citrate buffer, and place it at room temperature for 1-3h; then the mixture is treated with 100kDa Cut-off membrane ultrafiltration to remove unreacted polypeptides (citric acid reaction buffer as ultrafiltration liquid) to obtain HBc VLPs vaccines displaying each antigenic epitope polypeptide.

[0088] The result is as Figure 3A As shown, the size of the HBc-S monomer is about 27kDa, and after binding to the polypeptide, HBc-S-P is slightly larger than the HBc-S monomer, indicating that each polypeptide is bound to the surface of the HBc-S carrier protein; all the strips of HBc-S-P The bands all moved up, indicating that the binding efficiency of HBc-S to the polypeptide is extremely high.

[0089] Subsequently, the assembly of HBc-S-P was detected by t...

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Abstract

The invention provides a novel coronavirus antigen epitope and application thereof. A super computer is used for simulating S, E, M and N protein structures of the novel coronavirus, and novel coronavirus epitopes SEQ ID NO 1-38 are obtained through calculation. The epitopes have very good immunogenicity; an antibody generated by inducing a part of epitope polypeptide has the effect of neutralizing the novel coronavirus; the antigen epitope can be combined with antibodies in serum of novel coronavirus patients at home and abroad, has the potential of resisting various novel coronavirus strains, and also has the potential of identifying and applying different strains. The epitope provided by the invention can be used for (1) research and development of novel coronavirus vaccines and universal coronavirus vaccines such as MERS viruses and SARS viruses; and (2) preparing a novel coronavirus antibody, and further detecting and typing viruses and treating diseases caused by the viruses. Thenovel coronavirus antigen epitope has a wide application prospect in the aspects of prevention of coronavirus and propagation and outbreak thereof, and detection and diagnosis of virus strains.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to novel coronavirus epitopes and applications thereof. Background technique [0002] The novel coronavirus (SARS-CoV-2) was first reported in 2019, and its amino acid sequence has 94.6% homology with severe respiratory distress syndrome coronavirus (SARS-CoV). Similar to SARS, severe cases can lead to respiratory failure. However, due to the complex composition of SARS-CoV inactivated vaccines, most of the induced immune responses are non-protective, and the vaccine has biological safety hazards in the production process; attenuated vaccines have potential safety risks. The new vaccine based on the protective epitope has the characteristics of clear mechanism of action and good safety. Therefore, the protective antigenic epitope against SARS-CoV-2 can be obtained through rapid and high-throughput screening, and based on this, it can be designed and developed New epitope vaccines with g...

Claims

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Application Information

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IPC IPC(8): C07K14/165A61K39/215A61P31/14
CPCC07K14/005A61K39/12A61P31/14C12N2770/20022C12N2770/20034C07K2319/00
Inventor 刘瑞田卢帅谢喜秀
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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