Helicobacter pylori epitope vaccine, design method thereof, preparation method thereof and application thereof

A Helicobacter pylori epitope vaccine technology, applied in the field of biomedicine, can solve problems such as inability to effectively inhibit urease activity

Inactive Publication Date: 2011-08-17
CHINA PHARM UNIV
View PDF1 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, experimental studies have shown that anti-urease antibodies e

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
  • Helicobacter pylori epitope vaccine, design method thereof, preparation method thereof and application thereof
  • Helicobacter pylori epitope vaccine, design method thereof, preparation method thereof and application thereof
  • Helicobacter pylori epitope vaccine, design method thereof, preparation method thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0061] Example 1: Molecular structure design of Helicobacter pylori epitope vaccine CTB-UA

[0062] CTB forms a pentameric structure and combines with ganglioside GM1, which is very important for CTB to function as a mucosal immune adjuvant. Experimental studies have shown that when exogenous proteins are coupled to the C-terminus of CTB, the fusion protein can form a pentameric structure to the greatest extent, and has the strongest binding activity with ganglioside GM1. Therefore, in the design of the new Hp multi-epitope vaccine, the urease multi-epitope peptide (UE) was fused to the C-terminus of CTB to maintain the mucosal immune adjuvant activity of CTB to the greatest extent. Therefore, in the molecular structure design of Hp epitope vaccine CTB-UA, the urease A subunit epitope (UreA 183-203 ) is fused to the C-terminus of CTB.

[0063] In the design and research of enterotoxigenic Escherichia coli (ETEC) vaccine LTB-ST, it was shown that when 7 amino acids (DPRVPSS) ...

example 2

[0065] Example 2: Construction of recombinant expression vector pETCUA (containing fusion gene CTB-UA)

[0066] (1) Urease A subunit epitope UreAB 183-203B PCR synthesis of gene sequence

[0067] Primer P1-UreAB was designed using Primer Premier V5.0 183-203B and P2-UreAB 183-203B . Primer P1-UreAB 183-203B and P2-UreAB 183-203B The sequence and characteristics are as follows:

[0068] Primer P1-UreAB 183-203B :C C;

[0069] Primer P2-UreAB 183-203B :

[0070] Primer P1-UreAB 183-203The sticky end (C) with Kpn I restriction site at the front end and the sticky end (C) with Xho I restriction site at the back end are marked in black bold font; primer P2-UreAB 183-203 The sticky end with Xho I restriction site at the front end (TCGAG), and the sticky end with Kpn I restriction site at the back end (GGTAC), are marked in bold black font; primer P1-UreAB 183-203 and P2-UreAB 183-203 The middle part is the complementary urease A subunit epitope UreAB 183-203 an...

example 3

[0081] Example 3: Prokaryotic expression of Hp epitope peptide fusion protein CTB-UA

[0082] Transform the correct recombinant expression plasmid pETCUA into E.coli BL21(DE3) strain. On the pre-prepared LB plate containing 50 μg / mL Amp, inoculate the loop-streaked genetically engineered strain BL21(DE3) / pETCUA, place it upside down in a 37°C incubator, and after cultivating overnight for 12-16 hours, pick a single colony and inoculate it on a plate containing In LB medium with 50 μg / mL Amp, culture at 37°C, 180 rpm, for 12-16 hours. Inoculate the recombinant bacteria with 2% inoculum in LB medium containing 50 μg / mL Amp, shake overnight at 37°C and 180 rpm, and inoculate the bacterial solution at 1% inoculum in LB liquid culture containing 50 μg / mL Amp the next morning After shaking the flask at 37°C and 180rpm for 3 hours in the medium, IPTG was added to make the final concentration reach 1mmol / L, and the expression was induced at 37°C and 180rpm. The empty vector strain BL...

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 provides a helicobacter pylori epitope vaccine. The active constituent of the helicobacter pylori epitope vaccine is a polypeptide and mainly consists of a cholera toxin B subunit and a B cell antigen epitope from a urease A subunit. A preparation method of the helicobacter pylori epitope vaccine comprises the following steps of: synthesizing the nucleotide sequence of the B cell antigen epitope from the urease A subunit by a PCR (polymerase chain reaction) technology, coupling the nucleotide sequence with the gene sequence of the cholera toxin B subunit to form into a fusion gene, and expressing the fusion gene in the escherichia coli by an expression vector to obtain the fusion protein of the epitope vaccine by means of protein purification. The helicobacter pylori epitope vaccine can be used for inducing the human body to generate an epitope specific antibody which is higher in titer to the urease. In the biologic medical field, the epitope vaccine can be used for preventing and curing the relevant diseases caused by the helicobacter pylori infection, thereby being great in economic benefit and social benefit.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a Helicobacter pylori epitope vaccine and its design, preparation method and application. Background technique [0002] Helicobacter pylori (Hp) is an important pathogenic factor of chronic gastritis, peptic ulcer and duodenal ulcer, and is closely related to gastric cancer. The infection rate of Helicobacter pylori is very high. The Hp infection rate of the world population exceeds 50%, and the Hp infection rate of the Chinese population is 58.07%. It is clustered in families and the situation is even more severe. The current clinical treatment of Hp infection is mainly based on "triple" drug therapy (proton pump inhibitors, antibiotics, and bismuth), which has many disadvantages: such as toxic and side effects of drugs, increase of Hp drug-resistant strains, and repeated infections after drug withdrawal. Therefore, the "triple" drug therapy is not suitable for large-scal...

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): A61K39/106A61P31/04C07K19/00C12N15/62C07K14/435C12N15/09
Inventor 奚涛郭乐邢莹莹何赟绵王学全
Owner CHINA PHARM UNIV
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