VLP vaccine of recombinant HPV-58 L1 and preparation method thereof

A technology of vaccines and combination methods, applied in botany equipment and methods, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of large protein loss, difficulty of target protein, and difficult application in large-scale production

Active Publication Date: 2014-08-20
BEIJING HEALTH GUARD BIOTECH
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because most of the HPV L1 protein expressed by E. coli loses its natural conformation, it cannot produce protective antibodies against HPV.
Or, although the above proteins can be purified by inclusion bodies, renaturation and other steps can also be used to obtain HPV VLPs, but in the renaturation process, the protein loss is large and the yield is low, so it is difficult to apply in large-scale produc

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • VLP vaccine of recombinant HPV-58 L1 and preparation method thereof
  • VLP vaccine of recombinant HPV-58 L1 and preparation method thereof
  • VLP vaccine of recombinant HPV-58 L1 and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment l

[0072] Embodiment 1: have the construction of the engineering bacterium of HPV58 L1 sequence 2

[0073] 1、 The full-length HPV58 L1 gene was synthesized by GENEWIZ Biotechnology Co., Ltd. (GENEWIZ). Nucleotide sequence SEQ ID NO: 1, which is derived from GeneBank, the sequence number is GenBank: M14119.1.

[0074] 2. A template for PCR reaction containing the gene fragment of SEQ ID NO:1. With the forward primer sequence: 5'- CGCGGA TCCGGA AAGGAAGATC CATTAAATAA A -3'; a restriction endonuclease AccIII site was introduced at the 5' end of the H4 structure, and the sequence of the AccIII site was TCCGGA. The downstream contains the XhoI endonuclease site, the reverse primer sequence: 5'-GCTCTCCTCGAG TTA GGGCTTTGCT TTAAGGCCTG A -3', its 5' end introduces the restriction endonuclease XhoI site, and the XhoI site sequence is CTCGAG. HPV58B was amplified by PCR reaction.

[0075] 3. A template containing the gene fragment of SEQ ID NO: 1 for PCR reaction. To contain the intro...

Embodiment 2

[0080] Embodiment 2: Construction of engineering bacteria with HPV58 L1 sequence 4

[0081] 1. The target gene fragment of the full-length HPV58 L1 gene was purchased from the waste of clinical cell samples containing wild-type HPV58 virus from the Gynecology Clinic of Beijing Anzhen Hospital. Nucleotide The sequence is SEQ ID NO: 3 (GenBank: FN870689.1).

[0082] 2. A template containing the gene fragment of SEQ ID NO: 3 for PCR reaction. With the forward primer sequence: 5'- CGCGGATCCGGA AAGGAAGATC CATTAAATAA A -3'; a restriction endonuclease AccIII site was introduced at the 5' end of the H4 structure, and the sequence of the AccIII site was TCCGGA. The downstream contains the XhoI endonuclease site, the reverse primer sequence: 5'-GCTCTCCTCGAG TTA TGCACGGGTA GTAGGGGCTG A -3', its 5' end introduces the restriction endonuclease XhoI site, and the XhoI site sequence is CTCGAG. After PCR reaction, HPV58B was amplified.

[0083] 3. A template containing the gene fragmen...

Embodiment 3

[0088] Embodiment 3: Construction of engineering bacteria with HPV58 L1 sequence 6

[0089] 1、 The full-length HPV58 L1 gene was synthesized by GENEWIZ Biotechnology Co., Ltd. (GENEWIZ). Nucleotide sequence SEQ ID NO: 5, which is derived from GeneBank, the sequence number is GenBank: FN870694.1.

[0090] 2. A template containing the gene fragment of SEQ ID NO: 3 for PCR reaction. With the forward primer sequence: 5'- CGCGGATCCGGA AAGGAAGATC CATTAAATAA A -3'; a restriction endonuclease AccIII site was introduced at the 5' end of the H4 structure, and the sequence of the AccIII site was TCCGGA. The downstream contains the XhoI endonuclease site, the reverse primer sequence: 5'-GCTCTCCTCGAG TTA TGCACGGGTA GTAGGGGCCG A -3', its 5' end introduces the restriction endonuclease XhoI site, and the sequence of the XhoI site is CTCGAG. PCR reaction, amplified to obtain HPV58B.

[0091]3. A template containing the gene fragment of SEQ ID NO: 3 for PCR reaction. To contain the introd...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Diameteraaaaaaaaaa
The average particle sizeaaaaaaaaaa
Login to view more

Abstract

The invention provides a novel polynucleotide gene fragment coding recombinant HPV-58 L1 protein, a vector containing the gene fragment, a host cell including the vector, HPV-58 L1 protein pentamer or VLP translated and expressed by the gene fragment and a vaccine against HPV-58 infection composed by the HPV-58 L1 protein pentamer or VLP.

Description

technical field [0001] The present invention relates to the prevention and / or treatment of human papilloma virus infection. More specifically, the present invention relates to a recombinant human papillomavirus type 58 L1 protein, pentamers and VLPs composed of it, a vaccine containing the protein and its role in preventing HPV58 type virus infection, especially in preventing Use in cervical cancer diseases caused by HPV58 virus infection. Background technique [0002] Human Papillomavirus (HPV) is a DNA virus transmitted through close contact. In human tissues, HPV mainly infects skin and mucous membrane tissues. HPV DNA is divided into three categories according to the carcinogenicity of the virus: (1) Low-risk HPVs, including HPV6, 11, 40, 42, 43, 44, 54, 61, 70.72.51, mainly cause benign exophytic warts, cervical Intraepithelial neoplasm (cervical intraepithelial neoplasm, CIN) (2) high-risk HPV, including HPV 16, 18, 31, 33, 35, 39, 43, 51, 52, 56, 58, 59, 68, 73, 82...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K14/025C12N15/37C12N15/70C12N15/81C12N1/21C12N1/19A61K39/12A61P31/20
CPCA61K39/00C07K14/005C12N2710/20022C12N2710/20023C12N2710/20034
Inventor 刘永江陈小江陈林盖大海许铮曹科陈建平潘勇昭银飞阮芳勇
Owner BEIJING HEALTH GUARD BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap