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Preparation method of herpes zoster vaccine

A technology of herpes zoster and herpes zoster virus, which is applied in the field of virus vaccines, can solve problems such as potential safety hazards, unsatisfactory immune effects, and risks in the production process, so as to improve similarity, improve the level of glycosylation modification, and enhance immunity active effect

Pending Publication Date: 2019-09-17
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Live attenuated vaccines induce the body to form antibodies at the cost of low toxicity. The immune effect is not ideal and there are safety hazards
Inactivated vaccines only inactivate the virus during the use stage, and infectious viruses are still used in the production process, and there are risks in the production process

Method used

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  • Preparation method of herpes zoster vaccine
  • Preparation method of herpes zoster vaccine
  • Preparation method of herpes zoster vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Bioinformatics software was used to obtain three genes: the glycosylation site region gene of envelope glycoprotein I of herpes zoster virus, the gene of the outer envelope domain of envelope glycoprotein E, the gene encoding the glycine-linked peptide chain, And codon-optimized according to yeast codon preference.

[0032] The three genes are arranged in the following order: the envelope glycoprotein E extra-envelope domain gene is located after the signal peptide, followed by the encoding gene containing 35 glycine-linked peptide chains, followed by the envelope glycoprotein of the herpes zoster virus In the glycosylation site region gene of I, restriction enzyme sites are designed at both ends of the fusion gene: EcoRI and XhoI, for the recombination of the target gene into the pPICZαA expression vector; chemical synthesis of the target gene with EcoRI and XhoI sites , recombined with yeast expression vector pPICZαA and used for transformation of Pichia pastoris X33 ...

Embodiment 2

[0037] Bioinformatics software was used to obtain the gene of the outer envelope domain of envelope glycoprotein E, and the gene of the glycosylation site region of envelope glycoprotein I of herpes zoster virus, and directly connect them into fusion genes, and according to the yeast codon preference Perform codon optimization. Restriction enzyme sites were designed at both ends of the fusion gene: EcoRI and XhoI, for the recombination of the target gene into the pPICZαA expression vector; the fusion gene with EcoRI and XhoI sites was chemically synthesized, recombined with the yeast expression vector pPICZαA and used for The red yeast X33 strain was transformed, screened and identified, and then cultured in large quantities, and then the highly glycosylated herpes zoster virus envelope protein was secreted and expressed by methanol induction. The positive yeast single colony was inoculated in BMGY medium at 30°C, 250- 400rpm, cultivate to OD600=2-6; expand and inoculate in BS...

Embodiment 3

[0039] Use bioinformatics software to obtain the target genes, including the glycosylation site region gene of envelope glycoprotein I of herpes zoster virus, the gene of the outer envelope domain of envelope glycoprotein E, and the connecting peptide chain containing 50 glycines coding gene. Wherein, the glycine-linked peptide chain is: GGGGGAGGGG GSGGGGSGGG GGAGGGGGAG GGGGSGGGGG GGAGGGGGAG GGGGSGGGG The glycine chain used to connect the two polypeptide regions contains 50 glycines, and the short glycine peptides are spaced by alanine and serine.

[0040]The three genes are arranged in the following order: the glycosylation site region gene of the envelope glycoprotein I of the herpes zoster virus is located after the signal peptide, followed by the coding gene containing 50 glycine-linked peptide chains, and finally the envelope sugar. Protein E envelope domain gene, three gene groups are fusion genes, and restriction enzyme sites are designed at both ends of the fusion ...

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Abstract

The invention belongs to the field of virus vaccines and discloses a preparation method of a herpes zoster vaccine. Through combination use of a glycoprotein E envelope outer structural domain of the herpes zoster and a glycosylation locus area of envelope glycoprotein I, the herpes zoster vaccine is established, the envelope glycoprotein I, interacting with glycoprotein E of the herpes zoster, on a herpes zoster virus envelope, of the herpes zoster virus assists the glycoprotein E envelope outer structural domain of the herpes zoster in preparation of the herpes zoster vaccine, the glycosylation modification level of a vaccine protein complex is improved, the similarity of the vaccine protein complex and a natural virus envelope is improved, and the immunocompetence of the herpes zoster vaccine is improved.

Description

technical field [0001] The invention belongs to the field of virus vaccines, and particularly relates to a preparation method of a herpes zoster vaccine. Background technique [0002] Vaccines are the most economical and effective means to prevent and control infectious diseases. From the earliest vaccination with vaccinia to the current variety of vaccines, they play an important role in protecting human health. [0003] There are three main types of vaccines currently in use: live attenuated vaccines, inactivated vaccines, and genetically engineered vaccines. Live attenuated vaccines induce the body to form antibodies at the expense of low toxicity, and the immune effect is not ideal and has potential safety hazards. Inactivated vaccines only inactivate the virus in the use stage, and infectious viruses are still used in the production process, and there are risks in the production process. The genetic engineering vaccine induces the human body to form antibodies by hete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61P31/22C12N15/62C12N15/85C12N15/81C07K19/00
CPCA61K39/12A61P31/22C07K14/005C12N2710/16722C12N2710/16734C07K2319/00
Inventor 刘宝全李林李贵兴李春斌王剑锋权春善
Owner DALIAN NATIONALITIES UNIVERSITY
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