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Helicobacter pylori compound antigen, antibody, preparation method and application

A technology of Helicobacter pylori and Helicobacter pylori, which is applied in the field of biomedicine, can solve the problems of poor colonization inhibition effect and the effect needs to be improved, and achieve the effects of optimizing amino acid sequence, increasing potency, increasing concentration and purity

Active Publication Date: 2022-06-28
YOULIKANG (JIANGSU) BIOPHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this tetravalent antigen is mainly aimed at the toxin factor and HpaA of Helicobacter pylori, and has a poor effect on the colonization inhibition of some Helicobacter pylori that does not completely or does not rely on HpaA for adhesion, so its effect in treating different Helicobacter pylori infections still needs to be improved

Method used

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  • Helicobacter pylori compound antigen, antibody, preparation method and application
  • Helicobacter pylori compound antigen, antibody, preparation method and application
  • Helicobacter pylori compound antigen, antibody, preparation method and application

Examples

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Embodiment 1

[0048] This embodiment provides an Hp pentavalent composite antigen, which is composed of five antigen proteins, urease (UreB), vacuolar cytotoxin (VacA), cytotoxic gene A (CagA), adhesin HpaA and adhesin BabA; The amino acid sequences of the five antigenic proteins are respectively shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4 and SEQ ID No.5 after optimization; wherein, the sequence optimization includes The code preference of Escherichia coli and the sequence conservation of Helicobacter pylori are optimized, and the polypeptide sequence CTB is added to the N-terminus or C-terminus of the HpaA antigen protein. The CTB sequence is shown in SEQ ID No. 11; Add a protein expression tag to the C-terminus.

[0049] The specific preparation method is as follows:

[0050] Expression of S1 and Hp complex antigens

[0051] The nucleotide sequences corresponding to the five antigen proteins of urease (UreB), vacuolar cytotoxin (VacA), cytotoxin gene A (CagA), adhesin H...

Embodiment 2

[0069] This example provides an anti-Helicobacter pylori yolk antibody. The pentavalent composite antigen prepared in Example 1 is used to induce immunization of hens, and then a yolk antibody containing anti-five Hp antigens is obtained. The specific preparation steps are as follows:

[0070] (1) five kinds of Hp combined antigens are emulsified with adjuvant; the adjuvant is Freund's complete adjuvant;

[0071] (2) Immunization of laying hens: 10-40-week-old healthy laying hens are selected for isolation and rearing, and subcutaneous injection is carried out. Each hen is injected with 2-6 injection points. 80μg~200μg / chicken is preferred, 100μg / chicken is used in this example, after the first immunization, booster immunization is performed once every two weeks;

[0072] (3) collecting eggs produced by the last immunization, namely obtaining eggs containing yolk antibodies against five Hp antigens;

[0073] (4) separating and purifying the egg containing the egg yolk antibo...

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Abstract

The present invention provides a Helicobacter pylori composite antigen, which is composed of five full-length antigen proteins of urease (UreB), vacuolar cytotoxin (VacA), cytotoxin gene A (CagA), adhesin HpaA and adhesin BabA or partial sequences; the amino acid sequences of the five antigenic proteins are respectively shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4 and SEQ ID No.5. The composite antigen provided by the present invention has higher immunogenicity, and the anti-Hp yolk antibody prepared by the composite antigen has a stronger activity of neutralizing Hp, and can efficiently block the colonization of Hp and inhibit the activity of Hp at the same time. Effectively prevent and treat gastrointestinal diseases caused by Hp infection.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and particularly relates to a Helicobacter pylori composite antigen, an antibody, a preparation method and an application. Background technique [0002] Helicobacter pylori (Hp) is a spiral-shaped, Gram-negative microaerophilic bacillus that requires harsh growth conditions. It is the only microbial species found so far that can survive in the human stomach. It was first isolated in 1982. H. pylori can cause a variety of digestive tract diseases and even gastric cancer. The global infection rate exceeds 50%. The continuous increase of its drug resistance leads to a continuous decline in the eradication rate, which in turn promotes the occurrence of a large number of intractable infections, which seriously threatens human health. [0003] H. pylori continuously colonizes human gastric and duodenal mucosa, and causes vacuolar degeneration, abnormal proliferation and differentiation of gastric ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/80C07K14/195C07K1/22C12N15/55C12N15/31C12N15/70C07K16/12C07K16/02A61K39/40A61P31/04
CPCC12N9/80C07K14/195C12N15/70C07K16/121C07K16/02A61P31/04C12Y305/01005C12N2800/22C07K2317/11C07K2317/92C07K2317/35A61K2039/505Y02A50/30
Inventor 唐小军张秀芳
Owner YOULIKANG (JIANGSU) BIOPHARMACEUTICAL CO LTD
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