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Recombinant vector, recombinant protein and virus-like particle of human papilloma virus 16-type epitope chimeric L1 as well as preparation and application thereof

A technology of human papillomavirus and recombinant vector, which is applied in the direction of virus/bacteriophage, virus, virus peptide, etc., can solve the problem of weak cellular immune response, and achieve the effect of strong cellular immune response, plump shape, and uniform particle size

Inactive Publication Date: 2020-11-17
JINLIN MEDICAL COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the cellular immune response induced by such vaccines is weak

Method used

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  • Recombinant vector, recombinant protein and virus-like particle of human papilloma virus 16-type epitope chimeric L1 as well as preparation and application thereof
  • Recombinant vector, recombinant protein and virus-like particle of human papilloma virus 16-type epitope chimeric L1 as well as preparation and application thereof
  • Recombinant vector, recombinant protein and virus-like particle of human papilloma virus 16-type epitope chimeric L1 as well as preparation and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0043] main experimental materials

[0044] 1. Plasmids, cells and animals

[0045] 293FT cells were purchased from BioVector NTCC; pET28a plasmid was purchased from TaKaRa; plasmid pShell16 was purchased from addgene; pcDNA3.1-Luc+ Purchased from BioVector NTCC; Balb / c mice were purchased from the Animal Center of Bethune School of Medicine, Jilin University.

[0046] 2. Reagents, enzymes, medicines and instruments

[0047] BamH I and Xho I endonucleases were purchased from TaKaRa Company; Escherichia coli BL21(DE3) and DH5α competent cells were purchased from Beijing Dingguo Biotechnology Co., Ltd.; plasmid mini-extraction kit and agarose gel DNA recovery kit were purchased from from Tiangen Company; PVDF membrane, ECL chemiluminescence kit, and HRP-labeled goat anti-mouse IgG were purchased from Beyond Company; mouse anti-HPV16 L1 monoclonal antibody was purchased from Abcam Company in the United Kingdom; DMEM and 1640 medium were purchased from GIBCO Company in the United...

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Abstract

A virus-like particle system containing a human papilloma virus 16 type epitope chimeric DNA fragment is constructed; firstly, an epitope chimeric main capsid protein L1 DNA fragment containing humanpapilloma virus 16 type E749-57 is connected to a plasmid pET28a to construct a recombinant plasmid pET28a-16L1-E749-57, and then is transferred into escherichia coli for culture to obtain recombinantprotein; and self-assembling is carried out, such that virus-like particles (VLPs) are obtained. The assembled epitope chimeric virus-like particles can stimulate humoral and cellular immune responses at the same time.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant vector containing human papillomavirus type 16 epitope chimeric L1, a recombinant protein, a virus-like particle and its preparation and application. Background technique [0002] Cervical cancer is one of the most common malignant tumors in women, and its incidence rate ranks second in female malignant tumors, and its incidence tends to be younger. According to the World Health Organization, in 2018, there were 570,000 new cases of cervical cancer and 311,000 deaths worldwide, seriously endangering women's health. At present, research on cervical cancer preventive vaccines is relatively successful. The HPV vaccines marketed internationally are the quadrivalent vaccine Gardasil-4 (Gardasil-4) and the nine-valent vaccine Gardasil-9 (Gardasil-9) produced by Merck. As well as GlaxoSmithKline’s bivalent vaccine Cervarix, all three vaccines use the HPV L1 capsid protein sel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/70C07K19/00A61K39/12A61P31/20A61P35/00
CPCC07K14/005C12N15/70A61K39/12A61P31/20A61P35/00C07K2319/00C12N2710/20022C12N2710/20023C12N2710/20034
Inventor 刘微孙美艳郭健张磊许会静王皓
Owner JINLIN MEDICAL COLLEGE
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