Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Recombinant plasmid vaccine for treating hepatitis B and composition thereof

A technology of recombinant plasmids and recombinant vaccines, which is applied in the field of biomedicine and can solve problems such as the unsatisfactory immune effect

Active Publication Date: 2011-11-09
BEIJING KAWIN TECH SHARE HLDG
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Although the preliminary clinical trial results of DNA vaccine are satisfactory, its immune effect has not yet reached the ideal level. Therefore, how to enhance the immune effect has become the key to DNA vaccine research, especially how to ensure that DNA vaccine can simultaneously obtain a stronger effect. The immune effect of HBsAg and HBcAg

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
  • Recombinant plasmid vaccine for treating hepatitis B and composition thereof
  • Recombinant plasmid vaccine for treating hepatitis B and composition thereof
  • Recombinant plasmid vaccine for treating hepatitis B and composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Example 1 Construction of recombinant plasmid pRec2.0-preS2S-C (abbreviated as pS2S-C)

[0119] (1) First construct the recombinant plasmid vector backbone pOE-EKS, whose nucleotide sequence is the nucleotide sequence indicated by SeqNo.1:

[0120] Using pDRVISV1.0 as a template, Primer1 and Primer2 as primers, among which primer 2 is a primer phosphorylated at the 5' end, amplified to obtain a replicon region (Ori) with a size of 748bp, and introducing EcoRI, KpnI and SwaI restriction sites at the same time point;

[0121] Primer1: 5'-GGAATTCGGGGTACCATTTAAATTTGAACGTTCGCAAtATGTGAGCAAAAGGCCAGC-3'

[0122] Primer2: 5'-CGGCGCGCGCCGAAAACGACGATTGCGAACGTTCAACCCGTAGAAAAAGATCAAAGG-3'

[0123] Using pDRVISV1.0 as a template, using Primer3 and Primer4 as primers, among which primer 3 is a primer phosphorylated at the 5' end, amplified to obtain a Kanna resistance marker gene (Kan) with a size of 1056bp, and introducing an EcoRI restriction site ;

[0124] Primer3: 5'-tcgtcgt...

Embodiment 2

[0145] Example 2 Construction of recombinant plasmid pRec2.0-C-preS2S (pC-S2S)

[0146] Using the pRec2.0-PreS2S constructed in Example 1 as a template, using Primer11 and Primer12 as primers, amplify the PreS2S gene with a size of 873bp, and pass HindIII+XbaI double digestion and HindIII+XbaI double digestion vector pcDNA3 .1 connect, construct and obtain pcDNA3.1-PreS2S;

[0147]Primer11: 5'-CCCAAGCTTGCCGCCACCATGCAGTGGAACTC-3'

[0148] Primer 12: 5'-GCTCTAGAATCAGATGTAAACCCAC-3'

[0149] The PreS2S expression cassette was excised from the plasmid pcDNA3.1-PreS2S by BglII+PvuII double enzyme digestion, and cloned into pRec2.0-C which was digested by BglII+EcoRV double enzymes, and the recombinant plasmid pRec2.0-C-PreS2S (pC -S2S).

Embodiment 3

[0150] Example 3 Construction of recombinant adjuvant plasmid pmIL12

[0151] (1) Ligate the synthetic gene mP35 with the HindIII+XbaI double-digested vector pcDNA3.1 through HindIII+XbaI double-digestion to construct and obtain the recombinant plasmid pcDNA3.1-mP35;

[0152] (2) The mP35 gene expression cassette was excised from the plasmid pcDNA3.1-mP35 by BglII+PvuII double digestion, and cloned into the plasmid pRec2.0-PreS2S which was digested by BglII+EcoRV to obtain the recombinant plasmid pRec2.0-PreS2S -mP35;

[0153] (3) Ligate the synthetic gene mP40 with the HindIII+XbaI double-digested vector pcDNA3.1 through HindIII+XbaI double-digestion to construct and obtain the recombinant plasmid pcDNA3.1-mP40;

[0154] (4) The mP40 gene expression cassette was cut out by SalI+PvuII double enzymes, cloned into the plasmid pRec2.0-PreS2S-mP35 cut by SalI+SwaI double enzymes, and the recombinant plasmid pRec2.0-PreS2S-mIL12 was obtained;

[0155] (5) Digest the plasmid pRec2...

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

No PUM Login to View More

Abstract

The invention relates to a recombinant plasmid DNA (deoxyribonucleic acid) vaccine which simultaneously carries a hepatitis B surface antigen protein-coding gene and a hepatitis B core antigen protein-coding gene. The constructed recombinant plasmid DNA vaccine comprises two target protein expression cassettes, wherein one target protein expression cassette is an optimized target protein expression cassette, and the two genes are separated by one target protein expression cassette. The invention also relates to a double plasmid DNA vaccine which comprises the recombinant plasmid DNA vaccine provided by the invention and a recombinant interleukin 12 adjuvant plasmid. The recombinant plasmid DNA vaccine and the composition thereof constructed by the invention can be used for preparing medicaments for preventing and treating hepatitis B.

Description

technical field [0001] The invention relates to a recombinant plasmid DNA vaccine for treating hepatitis B disease and its composition, belonging to the field of biomedicine. Background technique [0002] Hepatitis B seriously endangers human health, and there is no effective treatment at present. The main reason for the chronicity of hepatitis B is that the body lacks persistent specific cellular immunity and humoral immunity after hepatitis B virus (HBV) infection. [0003] HBV coat protein is composed of large protein LS (S1S2S antigen, encoded by pre-S1-pre-S2-S gene), medium protein MS (S2S antigen, encoded by pre-S2-S gene), small protein S (S antigen, encoded by S gene coding) composed of three molecules, among which the pre-S2 antigen has a small molecular weight and strong antigenicity, and involves the adsorption of hepatitis B virus to host cells. Therefore, the vaccine containing the pre-S2 antigen should be more effective in protecting the vaccinated. [0004] ...

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): A61K48/00A61K39/29A61K39/39C12N15/63C12N15/66C12N15/51A61P1/16A61P31/20
Inventor 李鼎峰刘勇周德胜张春丽
Owner BEIJING KAWIN TECH SHARE HLDG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com