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Nano vaccine taking SARS-CoV-2 virus S protein RBD region as antigen and preparation method of nano vaccine

A nano-vaccine, sars-cov-2 technology, which is applied in the preparation of the nano-self-assembled particle protein vaccine and the field of nano-self-assembled particle protein vaccine, and achieves the effects of high safety, good antigenicity and facilitating process expansion.

Inactive Publication Date: 2020-12-04
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

[0004] Although nano-self-assembled particle vaccine technology is expected to provide a new breakthrough for the prevention and treatment of SARS-CoV-2 virus, there are no relevant reports in the prior art

Method used

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  • Nano vaccine taking SARS-CoV-2 virus S protein RBD region as antigen and preparation method of nano vaccine
  • Nano vaccine taking SARS-CoV-2 virus S protein RBD region as antigen and preparation method of nano vaccine
  • Nano vaccine taking SARS-CoV-2 virus S protein RBD region as antigen and preparation method of nano vaccine

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0032] Example 1: Preparation of the nanometer self-assembled particle protein vaccine of the present invention.

[0033] Cell transfection plasmid preparation

[0034] 1) The gene sequence of the SARS-CoV-2 virus S protein RBD region (as shown in the sequence table SEQ ID NO.2) is optimized according to the codons of the human expression system and the whole gene is synthesized, and constructed on the pc3.1+ vector backbone .

[0035] 2) Construction of the plasmid The correct expression frame was verified by sequencing, and the E. coli seed bank of the target plasmid was established.

[0036] 3) Use the endotoxin removal plasmid kit to extract the SARS-CoV-2 virus S protein RBD region expression plasmid.

[0037] Protein expression conditions in different cell lines

[0038] 1. Protein expression in 293T cell system

[0039] The SARS-CoV-2 virus S protein RBD region expression plasmid was transiently transfected into 293T cells:

[0040] 1) Use a 10cm culture plate for ...

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Abstract

The invention discloses a nano vaccine taking an SARS-CoV-2 virus S protein RBD region as an antigen and a preparation method of the nano vaccine. An expression vector with an SARS-CoV-2 virus S protein RBD region gene sequence is transfected into 293T, 293i and CHO cell lines for antibody expression, and the required vaccine is obtained after protein extraction, ultrafiltration enrichment, antibody affinity chromatography purification, protein collection after purification, PBS dialysis and aluminum hydroxide adjuvant mixed incubation. The vaccine is a protein for expressing an amino acid sequence of the SARS-CoV-2 virus S protein RBD region, 24 monomers in a cell can be self-assembled into a sphere structure, and an SARS-CoV-2 virus S protein RBD peptide fragment subjected to fusion expression is displayed on the surface of a sphere. The vaccine has high neutralizing activity on the SARS-CoV-2 virus, and can effectively inhibit infection of the virus on vero cells. A toxicity attacking experiment shows that the vaccine can obviously and effectively protect the infection and damage of the SARS-CoV-2 to rhesus monkeys, and an excellent candidate vaccine is provided for the prevention and control of the current COVID-19 epidemic situation.

Description

technical field [0001] The invention belongs to the technical field of coronavirus vaccine preparation, in particular to a nanometer self-assembled granular protein vaccine with good neutralizing activity against SARS-CoV-2 virus. At the same time, the invention also relates to a preparation method of the nanometer self-assembled granular protein vaccine. Background technique [0002] In recent years, vaccine preparation technology has been continuously developed, and the newly emerged nano-self-assembled particle vaccine has the following advantages: fast preparation, simple purification process, easy to scale up, multiple antigen monomer polymerization display makes antigenicity higher, and is produced by human cell fermentation Higher security. For example, the technology of displaying the conserved antigenic sites of influenza virus on the surface of self-assembled nanoparticles to enhance vaccine antigenicity and safety has been widely used in the development of univer...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61P31/14C12N15/85C12N15/50
CPCA61K39/12A61P31/14C12N15/85C07K14/005C12N2800/107C12N2770/20022C12N2770/20034A61K2039/55505
Inventor 刘龙丁李恒郭磊郑惠文梁燕杨泽宁陈燕丽和占龙施海晶李菁
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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