Hen egg yolk antibody for resisting pyloric helicobacterium cytotoxin, its preparation and application

A technology of chicken egg yolk antibody and cytotoxin, which is applied in the field of egg yolk antibody product-anti-Helicobacter pylori cytotoxin chicken egg yolk antibody, to achieve the effect of easy industrialization

Inactive Publication Date: 2002-06-19
重庆和润实业(集团)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the prior art, there is no report on the method of immunizing healthy hens by using Helicobac...

Method used

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  • Hen egg yolk antibody for resisting pyloric helicobacterium cytotoxin, its preparation and application
  • Hen egg yolk antibody for resisting pyloric helicobacterium cytotoxin, its preparation and application

Examples

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Embodiment 1

[0022] Embodiment 1: The product of Helicobacter pylori cytotoxin yolk antibody with antibody activity can be obtained by the following steps: prepare Helicobacter pylori (HP) cytotoxin antigen, immunize healthy hens with HP toxin antibiotic antigen, and then collect the produced eggs, and then extract anti-HPTxIgY from the eggs laid.

Embodiment 2

[0023] Example 2: Preparation of HP cytotoxin antigen, isolate the toxin-producing HP strain, smear it on Brooke's agar, and culture it microaerobically at 37°C for 72 hours. After obtaining pure strains, expand it in Brooke's broth medium Culture T2h. Collect the culture supernatant, separate the toxin by ultrafiltration and chromatography, add complete Freund's adjuvant at a weight ratio of 1:1, and emulsify to obtain HP toxin antigen.

Embodiment 3

[0024] Embodiment 3: Construct the expression plasmid containing HP cytotoxin structural gene VacA or CagA, express in the Escherichia coli system, the expression product is through separation and purification, prepares HP vacuolating toxin (VacA) and cytotoxic toxin (CagA), by weight Mixed at a ratio of 1:1 as immune antigen, added an equal volume of Freund's adjuvant, emulsified and mixed to obtain HP toxin antigen.

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Abstract

The present invention relates to a chicken vitellus antibody (anti-HPTxIgY) with antibody activity for resisting pyloric helicobacterium cytotoxin, its preparation method and application in the preparation of medicine and health-care product for curing related diseases resulted from pyloric helicobacterium infection and preparation for detecting these related diseases. The above-mentioned antibody is undergone the process of IgY purity checking treatment. PAGE electrophoressi can be used for detecting purified vitallus antibody purity. Only one electrophoretic band is produced in the electrophoresis, and an UV spectrophotometer is used for determination so as to calculate the albumen concentration (mg/ml) which is (1.45XOD 280nm-0.74XOD 260nm)X dilute multiple. The above-mentioned method is as follows: preparing pyloric helicobacterium cytotoxin antigen, immunizing health hen with said antigen, collecting egg and extracting anti-HPTxIgY from the egg.

Description

(1) Technical field [0001] The invention relates to an egg yolk antibody product with antibody activity-anti-helicobacter pylori cytotoxin chicken egg yolk antibody, and relates to its preparation process and pharmaceutical application. (2) Technical background [0002] Helicobacter pylori (Helicobacter pylori, HP) is a pathogenic bacterium closely related to the occurrence of chronic gastritis, peptic ulcer and non-ulcer dyspepsia discovered in the past two decades. WHO lists it as a class I carcinogen. HP can produce a variety of protein molecules with cytotoxic activity. After colonization in the gastric mucosa, HP releases toxins such as VacA (vacuolar toxin) and CagA (cytotoxin-like protein) Necrosis of gastric mucosal epithelium and inflammation. Therefore, cytotoxin molecules play an important role in the pathogenic process of HP. At present, diseases related to HP infection, such as chronic gastritis, peptic ulcer, gastric cancer, etc. [0003] The treatment of HP...

Claims

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Application Information

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IPC IPC(8): A61K31/04C07K16/18C07K16/44C12P21/02C12Q1/04
Inventor 陈君雁杰
Owner 重庆和润实业(集团)有限公司
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