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Yolk antibody specific for helicobacter pylori and its preparation and application

An anti-Helicobacter pylori and yolk antibody technology, applied in application, antibody, food preparation, etc., can solve the problems of ecological imbalance and poor specificity of intestinal flora, and achieve the effect of easy industrialization, low cost and good specificity

Inactive Publication Date: 2005-03-16
上海立泰哲田生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are also a small number of reports on the preparation of IgY from HP immunized laying hens, but the antigens they use are HP whole bacterial antigens or crudely extracted antigens. The result is poor specificity. Group ecological imbalance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: the composite yolk antibody (anti--HPSlgY) of the specific protein antigen of anti-HP can be obtained through the following steps: (1) adopt genetic engineering method to prepare the specific protein antigen of HP; (2) immunize egg-laying hen: select Healthy egg-laying hens aged 10-40 weeks were reared in isolation and subcutaneously injected with 2-8 injection points for each hen. The antigen concentration for immunization was 100-500 μg / ml, and the same amount of Freund’s adjuvant was added. Each immunization dose of each hen is 100-600μg antigen, after the first immunization, booster immunization once every two weeks, a total of 8 times of immunization; (3) collect the eggs produced after the last immunization; (4) collect the eggs from the immunized egg yolk Antibody isolation and purification.

Embodiment 2

[0022] Example 2: Preparation of HP-specific protein antigens by genetic engineering recombination method: Gastric mucosal tissues were taken from patients with gastric ulcers positive for HP infection with a gastroscope, spread on HP selective culture Brookfield agar plates, and cultured in microaerobic at 37°C After 5 days, colonies were collected, HP DNA was extracted by conventional methods, UreB, Hsp60, CagA, and VacA genes of HP were amplified by PCR, the above four genes were cloned into E. coli expression plasmids, recombinant plasmids were transformed into E. coli, and recombinant E. coli was fermented. Bacillus, UreB, Hsp60, CagA, VacA proteins in Escherichia coli were purified by chromatography.

Embodiment 3

[0023]Example 3: Immune egg-laying hens: select 10-week-old healthy egg-laying hens for isolation and rearing, and inject subcutaneously into chickens. There are 2 injection points for each hen, and the antigen concentration for immunization is 100 μg / ml, and an equal amount is added Freund's adjuvant, each immunization dose of each hen is 100 μg of antigen, after the first immunization, booster immunization once every two weeks, a total of 8 times of immunization;

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Abstract

The invention discloses a yolk antibody specific for helicobacter pylori, its preparation and use thereof, wherein gene recombination HP specific proteantigens are employed to immune the oviposition hens, and composite vitelline antibodies (anti-HPSlgY) are extracted from the eggs. The invention can be applied to charge yolk powder containing composite vitelline antibodies into foodstuff or be used to prepare health food, can be used in the prevention and cure of diseases caused by Helicobacter pylori.

Description

technical field [0001] The present invention relates to an antibody and its application, in particular to the preparation of a composite yolk antibody against Helicobacter pylori specific antigen and its application in food, health food and medicine. technical background [0002] In 1982, Warren and Marshall isolated a new bacterium, now called Helicobacter pylori (HP), from the gastric biopsy tissue of a patient with duodenal ulcer, and proposed that it was peptic ulcer pathogenic factor. Fourteen years of research confirmed this hypothesis. In February 1994, at a special meeting hosted by the National Institutes of Health (NIH), an independent panel of experts declared peptic ulcer disease to be an infectious disease and recommended antibiotic therapy to replace the previous antacid therapy. [0003] Helicobacter pylori infection spreads all over the world, and the infection rate is high in economically underdeveloped areas, and increases with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23L33/135A61K39/395A61P1/04A61P31/04C07K16/02C07K16/12G01N33/563
Inventor 许春娣徐帆洪
Owner 上海立泰哲田生命科技有限公司