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Nucleic acid vaccine for preventing HPV (human papilloma virus) infection

A technology of HPV-L1 and nucleotide sequence, which is applied in the field of preventing HPV infection, HPVL1 microcircle DNA vaccine and its preparation, and achieves the effects of low industrial production cost, enhanced immune response, and simple preparation process

Inactive Publication Date: 2018-08-21
江苏润洁生物科技有限公司
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No minicircle DNA vaccine against HPV L1 capsid protein for cervical cancer prevention

Method used

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  • Nucleic acid vaccine for preventing HPV (human papilloma virus) infection

Examples

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Embodiment 1

[0042] Example 1 Preparation of HPV L1 microcircle DNA

[0043] 1. Reagent Preparation

[0044] 1) TB culture solution: 12 g of peptone, 24 g of yeast extract and 4 ml of glycerin were dissolved in 900 ml of water. After the components are dissolved, autoclave. Cool to 60°C, then add 100ml of a mixture of 170mmol / L KH2PO4 and 0.72mol / L K 2HPO4 that has been sterilized by autoclaving or filtered through a 0.22μm membrane, and the mixture is obtained.

[0045] 2) Minicle Induction Mix: Add 4ml of 1M NaOH and 200μl of 20% arabinose to 100ml of LB, mix and filter through a 0.22μm filter membrane.

[0046] 2. Preparation of recombinant microcircle DNA

[0047] 1) Designing and synthesizing the HPV-L1 recombinant fragment of the objective one with BglII and SalI restriction sites added at both ends to obtain the objective gene fragment;

[0048] 2) Digest the above target gene fragment and the ZY781 vector with BglII and SalI at the same time, and clone the target fragment into ...

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Abstract

The invention belongs to the field of biological medicine and particularly relates to an HPV (human papilloma virus) L1 minicircle DNA vaccine and an application of the vaccine to prevention of HPV infection. The HPV L1 minicircle DNA contains a nucleotide sequence of HPV 16 subtype L1 (major capsid protein). Meanwhile, the minicircle DNA has following advantages: 1), the minicircle DNA is not integrated into a host cell genome, cannot induce new mutation and is safer than a virus vector; 2), the minicircle DNA can be amplified massively in engineering bacteria, the preparation technology is simple, and industrial production cost is low; 3), exogenous gene expression is more stable and more efficient as compared with expression of traditional plasmid. The HPV L1 minicircle DNA has great significance for cervical cancer prevention when applied to vaccine preparation.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an HPV L1 microcircle DNA vaccine, a preparation method thereof, and an application in preventing HPV infection. Background technique [0002] Cervical cancer accounts for the second place among female gynecological malignancies, and has now become an important disease that threatens women's health. In China, nearly 50,000 people die from cervical cancer every year. The etiology of cervical cancer has always been valued by scholars at home and abroad. The key to cervical cancer is HPV infection. HPV is a spherical, double-stranded circular DNA virus belonging to the genus Papillomavirus of the family Papillomaviridae. HPV infects humans mainly through direct or indirect contact with contaminated objects or through sexual transmission. HPV infection has host specificity and tissue specificity, and can only infect damaged basal cells of human skin and mucous membranes. [...

Claims

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Application Information

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IPC IPC(8): C12N15/62C07K19/00A61K39/12A61P31/20
CPCA61K39/12A61K2039/53A61P31/20C07K14/005C07K2319/02C12N2710/20022C12N2710/20034
Inventor 庞以芹谢亦武吴必杰魏勇
Owner 江苏润洁生物科技有限公司
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