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Protein for resisting SARS-CoV-2 infection and vaccine

A sars-cov-2, protein technology, applied in the field of medicine, can solve the problems of prevention and treatment, no antiviral drugs, etc., and achieve the effect of convenient purification

Active Publication Date: 2021-02-02
WEST VAC BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the new coronavirus SARS-CoV-2 has caused tens of thousands of infections, and there is no effective antiviral drug for prevention and treatment. The development of a vaccine against the virus is very important for the prevention and treatment of the disease

Method used

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  • Protein for resisting SARS-CoV-2 infection and vaccine
  • Protein for resisting SARS-CoV-2 infection and vaccine
  • Protein for resisting SARS-CoV-2 infection and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Embodiment 1 prepares the protein of anti-SARS-CoV-2 infection of the present invention with insect baculovirus expression system

[0091] Vector construction: The recombinant protein produced by the insect baculovirus expression system mainly utilizes the S protein receptor binding region (RBD). The SARS-CoV-2 virus S protein is a protein located on the viral envelope. In order to simulate its secretion process, when constructing the S protein receptor binding region (RBD), a GP67 signal peptide was added to its N-terminal to help the protein The secretory expression of this signal peptide will be spontaneously excised by insect cells during the process of protein secretion. At the same time, in order to facilitate purification and increase the water solubility of the protein, a thioredoxin tag and an enterokinase (EK enzyme) cleavage site were also introduced into the sequence. The complete nucleotide sequence is shown in SEQ ID No.8 or SEQ ID No.10. The expression ...

Embodiment 2

[0095] Embodiment 2 prepares the protein of anti-SARS-CoV-2 infection of the present invention with CHO cell expression system

[0096] The recombinant protein vaccine produced by CHO cells is mainly aimed at the S protein receptor binding region (RBD). These fragments are gene synthesized according to codon preference, and polyhistidine is used as a purification tag (6His). The nucleotide sequence is shown in SEQ ID No.13. Then it is constructed on the high-expression vector pTT5 to express the precursor protein whose amino acid sequence is shown in SEQ ID No.14.

Embodiment 3

[0097] Example 3 Expression of the present invention's anti-SARS-CoV-2 infected protein in Escherichia coli

[0098] Use pET32a (Novagen Company) as the expression vector (plasmid map see Figure 14 ), which contains the T7 promoter, and is regulated by IPTG to regulate the transcription of the downstream target gene. The N-terminal of the expression product is fused with thioredoxin (Trx), and can be purified in one step by using metal chelate column affinity chromatography. In order to remove Trx after the target protein is purified, an enzyme cleavage site for enterokinase is added after Trx. The complete nucleotide sequence is shown in SEQ ID No.11.

[0099] The recombinant expression plasmids were expressed in Escherichia coli strain BL21(DE3), and the electrophoresis results showed that the size of the target protein was similar to that expected, and was identified by Western Blot. The content of the target protein can reach more than 30% of the total protein, and it ...

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Abstract

The invention relates to a protein for resisting SARS-CoV-2 infection and a vaccine, and belongs to the field of medicines. The problem of lack of effective prevention and treatment drugs for SARS-CoV-2 infection is solved. On one hand, the protein for resisting SARS-CoV-2 infection contains a structural domain combined with an angiotensin converting enzyme 2 receptor in an S protein of SARS-CoV-2, and the first amino acid arginine at the N end of the structural domain can be deleted. On the other hand, the vaccine for preventing and / or treating the SARS-CoV-2 infection contains the protein for resisting the SARS-CoV-2 infection, and pharmaceutically acceptable auxiliary materials or auxiliary components. According to the protein for resisting SARS-CoV-2 infection and the vaccine, the combination of the S protein of the SARS-CoV-2 and an ACE2 receptor of a host cell is blocked by inducing in-vivo generation of antibodies and other immune responses, so that a host is helped to resist coronavirus infection.

Description

technical field [0001] The invention relates to a protein and a vaccine against SARS-CoV-2 infection, and belongs to the field of medicine. Background technique [0002] SARS-CoV-2 is a new type of coronavirus named by the World Health Organization. The virus has an envelope, and the particles are round or oval, often pleomorphic, with a diameter of 60-140nm. Its genetic characteristics are significantly different from SARS-CoV and MERS-CoV, and it is a new branch of coronavirus that has not been found in humans before. Bats may be the natural host of SARS-CoV-2, and some studies suggest that pangolins may also be the animal source of the virus. At present, the new coronavirus SARS-CoV-2 has caused tens of thousands of infections, and there is no effective antiviral drug for prevention and treatment. The development of a vaccine against the virus is very important for the prevention and treatment of the disease. [0003] The main structural proteins of SARS-CoV-2 include ...

Claims

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Application Information

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IPC IPC(8): C07K14/165C07K19/00C12N15/50C12N15/62A61K39/215A61K39/39A61P31/14
CPCC07K14/005A61K39/12A61K39/39A61P31/14C12N2770/20022C07K2319/50C07K2319/21C07K2319/02C07K2319/35C12N2770/20034A61K2039/55505A61K2039/55577A61K2039/55583A61K2039/55566A61K2039/55544A61K2039/54A61K2039/575A61K39/215
Inventor 魏霞蔚逯光文王玮杨金亮杨莉李炯杨静云魏于全王震玲赵志伟沈国波
Owner WEST VAC BIOPHARMA CO LTD
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