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Tandem epitope polypeptide vaccine of novel coronavirus and application thereof

A coronavirus and epitope technology, applied in the direction of viral peptides, antiviral immunoglobulins, viruses, etc., can solve the problems of low response rate of the crowd, low immunogenicity, difficulty in overcoming virus immune escape, etc.

Pending Publication Date: 2021-12-17
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the immune protection produced by currently developed vaccines is limited, such as low immunogenicity, safety risks, low population response rate, and difficulty in overcoming viral immune escape

Method used

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  • Tandem epitope polypeptide vaccine of novel coronavirus and application thereof
  • Tandem epitope polypeptide vaccine of novel coronavirus and application thereof
  • Tandem epitope polypeptide vaccine of novel coronavirus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0167] Example 1 T / B cell epitope screening and tandem epitope polypeptide design based on the sequence and structure analysis of S protein RBD

[0168] By analyzing the sequence and structure of the S protein RBD of SARS-CoV-2, the present inventors predicted and analyzed the CD4+T / CD8+T cell epitope, linear / conformation B Cellular epitopes, structural features and key sites of interaction. After comprehensive analysis of the results, suitable T / B cell antigen epitopes were screened and determined, and these epitopes were concatenated with the universal Th epitope PADRE sequence, wherein the epitopes were connected by four glycines to ensure mutual non-interference.

[0169] Specifically, the inventors took the RBD region interacting with human ACE2 in the SARS-CoV-2 virus S protein as the analysis object, and determined the key sites in the RBD interacting with ACE2, such as figure 1 shown.

[0170] The present inventor also used Allele Frequency Net Database statistical a...

Embodiment 2

[0178] The preparation of embodiment 2 polypeptide

[0179] In this example, a fully automatic solid-phase peptide synthesizer was used to prepare the peptide LP2.

Embodiment 3

[0180] Example 3 The immune effect of LP2 polypeptide vaccine

[0181] In this example, the polypeptide LP2 prepared in Example 2 was used to immunize cynomolgus monkeys, and the immune effect of LP2 was evaluated.

[0182] Mix the polypeptide LP2 with an adjuvant (such as TiterMax) to prepare an immune preparation, immunize cynomolgus monkeys by subcutaneous multi-point injection, and 14 days after the second immunization, use the Bridging-ELISA method to measure the antibody titer, and measure the antiserum Blocking capacity of RBD binding to ACE2.

[0183]The neutralizing antibody was detected by the competitive ELISA method, and the specific determination method was as follows: 10 μg / mL ACE2 was coated overnight on the microtiter plate, and it was blocked before use. Dilute the anti-peptide serum (1:128, 1:64, 1:32, 1:16, 1:8 and 1:4) with sample dilution buffer to different degrees, and then mix the diluted antiserum with 12 μg / mL of Bio-RBD was incubated at 37°C for 1...

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Abstract

The invention discloses a tandem epitope polypeptide vaccine of a novel coronavirus and application of the tandem epitope polypeptide vaccine. Specifically, the invention provides a novel coronavirus pneumonia vaccine polypeptide based on analysis and research of RBD sequence and structural information of S protein of SARS-CoV-2, and the vaccine polypeptide comprises the following elements connected in series: a universal Th epitope sequence, a B cell epitope sequence and a T cell epitope sequence, wherein the B cell epitope and the T cell epitope have amino acid sequences from an RBM region of S protein of SARS-CoV-2. The invention further provides a vaccine composition containing the vaccine polypeptide and application of the vaccine composition. Experiments show that the vaccine polypeptide can enable cynomolgus monkeys to start relatively strong cell and humoral immunity, generates a neutralizing antibody for blocking the combination of RBD and ACE2, and can be used for preventing and treating novel coronavirus pneumonia.

Description

technical field [0001] The present invention relates to the fields of polypeptide drugs and polypeptide vaccines, in particular, to novel coronavirus tandem epitope polypeptide vaccines and applications thereof. Background technique [0002] The novel coronavirus pneumonia (Corona virus disease 2019, COVID-19) caused by the coronavirus SARS-CoV-2 is highly contagious. However, there is still a lack of clear and effective preventive and therapeutic drugs and measures for COVID-19, and supportive and symptomatic treatment is currently the main clinical approach. [0003] Establishing herd immunity against SARS-CoV-2 through vaccines is the ultimate way to control and block the COVID-19 epidemic. Various types of COVID-19 vaccines are currently in preclinical and clinical trials, including live attenuated vaccines, inactivated virus vaccines, recombinant virus vector vaccines, recombinant protein vaccines, DNA vaccines, RNA vaccines, and peptide vaccines. [0004] However, th...

Claims

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Application Information

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IPC IPC(8): C07K14/165A61K39/215A61P31/14
CPCC07K14/005A61K39/12A61P31/14C12N2770/20022C12N2770/20034A61K2039/5158A61K35/76C07K19/00A61K2039/575A61K2039/545C07K16/1003A61K39/4614A61K39/4615A61K39/4613A61K39/4611A61K39/464838A61K39/215A61K2039/70
Inventor 宫丽崑任进秦秋平朱维良龙益如徐志建孙建华刘婷婷靳广毅左建平黄蔚
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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