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High-stable anti-helicobacter pylori recombinant VacA specific egg yolk antibody and use thereof

A technology against Helicobacter pylori and Helicobacter pylori, which is applied in the direction of antibodies, antibacterial drugs, antibacterial immunoglobulin, etc., can solve the problems of high cost and complicated production, and achieve convenient material acquisition, simple preparation process, and enhanced antifreeze The effect of the ability to fuse

Inactive Publication Date: 2009-03-18
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are more complicated and costly to manufacture.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Preparation of specific IgY against Helicobacter pylori recombinant VacA

[0021] The purified recombinant VacA was mixed with an equal amount of complete Freund's adjuvant, fully ground and emulsified, and immunized with 300-day-old Lohmann hens, and vaccinated by multi-point injection in the breast muscle. The first immunization dose was 100ug / monkey; the second immunization was carried out 15 days later, the dose was 200ug / bird; the third immunization and booster immunization were carried out every month thereafter, and the dose was 300ug / bird. Egg collection began 1 week after the second immunization. The egg yolk liquid was first diluted with water and combined with chloroform organic precipitation method to extract IgY, then purified by conventional saturated ammonium sulfate method, sterilized by 0.22 μm filtration, analyzed for purity by 12% SDS-PAGE, and determined for protein content by Bradford method, and stored at -20°C.

Embodiment 2

[0022] Example 2 Preparation of highly stable anti-Helicobacter pylori recombinant VacA-specific yolk antibody

[0023] Take the purified anti-H. pylori recombinant VacA-specific egg yolk antibody, dilute it to pH 7.1 with pH 7.0 sodium dihydrogen phosphate buffer solution, and take 70 ml for later use; take another 30 ml of sucralfate, and dilute it with 70 ml The anti-H. pylori recombinant VacA-specific egg yolk antibody was fully mixed and stored at 4°C.

Embodiment 3

[0024] Example 3 Preparation of highly stable anti-Helicobacter pylori recombinant VacA-specific yolk antibody

[0025] Take the purified anti-Helicobacter pylori recombinant VacA-specific egg yolk antibody, dilute it to pH 7.1 with pH 7.0 sodium dihydrogen phosphate buffer solution, take 60 ml for later use; take another 40 ml of sucralfate, and dilute it with 60 ml The anti-H. pylori recombinant VacA-specific egg yolk antibody was fully mixed and stored at 4°C.

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PUM

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Abstract

The invention aims at providing a helicobacter pylori resistant recombined VacA specificity yolk antibody with convenient available material, simplicity, economical efficiency and high stability. Sucralfate with volume percent of between 30 and 50 percent is added to the antibody to serve as a protectant, thereby improving the stability of the specificity yolk antibody against acid and pepsin, strengthening the anti-freeze thawing capacity, and maintaining the activity of the yolk antibody at room temperature. In addition, the self protective function of the sucralfate on the gastric mucosa can also be brought into play, and the sucralfate can also be used for other specificity yolk antibodies. Regarding the preparation, the material of the antibody is conveniently available, simple and economical. Two kinds of liquids can be simply mixed according to the proportion for direct taking. The specificity yolk antibody can be used for treating helicobacter pylori infection.

Description

technical field [0001] The invention relates to an anti-helicobacter pylori recombinant VacA specific egg yolk antibody, in particular to a highly stable anti-helicobacter pylori recombinant VacA specific egg yolk antibody added with a protective agent and application thereof. Background technique [0002] Helicobacter pylori (Helicobacter pylori, Hp) is one of the bacteria with a high infection rate in the world. Children are susceptible to it. It is only discovered when symptoms appear. It is associated with chronic gastritis, peptic ulcer, gastric adenocarcinoma and mucosa-associated lymphoid tissue. The World Health Organization has listed it as the first type of carcinogen, and the prevention and treatment of Hp infection has attracted great attention from scholars at home and abroad. Antibiotics are commonly used to treat Hp infection clinically, but there are problems such as poor patient compliance, emergence of drug-resistant strains, and repeated infections. It is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02A61K39/40A61P31/04
Inventor 杨致邦郭丽媛
Owner CHONGQING MEDICAL UNIVERSITY
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