Production of hygrophilous monospermous bacterium main-protective antigen univalent and multivalent vitelline antibody and use in aquatic animal

A technology of Aeromonas hydrophila and protective antigen, applied in application, antibody, animal feed, etc., can solve the problem of non-compliance with the safety requirements of meat animal food, large irritation and toxic side effects, and specific antibody titer. Low problems, to achieve the effect of easy industrial production, easy preservation, and no safety problems

Inactive Publication Date: 2007-11-21
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent application: the patent application whose application number is 200510060325.X provides a kind of anti-Aeromonas hydrophila immunoglobulin preparation technology, but the shortcoming of this technology is: 1) the antigen adopted is whole bacteria inactivated antigen, The antigen particles are large, the components are complex, the antigen specificity is poor, and the specific antibody titer is low; 2) The disadvantage of using Freund's adjuvant (FCA) as an immune adjuvant is that ① FCA oil can cause tissue damage through the sarcolemma, Local nodules and aseptic granulomas are formed, which damage the meat quality; ②FCA contains mycobacteria, which can make the tuberculin test positive; ③FCA may have carcinogenic effects; ④Viscous, and the injection is laborious; ⑤Immune adjuvant Freund The adjuvant is too irritating and toxic to animals, which can lead to a significant drop in the egg production rate of laying hens, and does not meet the food safety requirements of meat animals, so it is banned from clinical use

Method used

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  • Production of hygrophilous monospermous bacterium main-protective antigen univalent and multivalent vitelline antibody and use in aquatic animal
  • Production of hygrophilous monospermous bacterium main-protective antigen univalent and multivalent vitelline antibody and use in aquatic animal

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Embodiment 1

[0064] A method for preparing the protective antigen monovalent yolk antibody of the Aeromonas hydrophila outer membrane protein, first obtaining the protective antigen-outer membrane protein from the Aeromonas hydrophila, and the outer membrane protein is composed of Prepared by the following method: inoculate bacteria in TSB medium containing 1% sodium chloride, shake at 150r / min at 30°C for about 24h, wash at 10000r / min, and wash 3 times with 0.02mol / L Tris-HCL (pH7.5) , resuspended in about 10mL of the above buffer, 200W ultrasonic crushing for about 6min, 7000r / min at 4°C, 10min to collect the supernatant, adding N-lauroylsarcosine sodium salt to make it final The concentration is 1.2% (W / V), mix well, and stand overnight at 4°C. 40 000r / min at 4°C for 40min, the lower precipitate is the main outer membrane protein, resuspended in distilled water to determine protein concentration by Bradford method, aliquoted, and stored at -20°C for later use. Then the obtained main ou...

Embodiment 2

[0075] The remaining steps and methods of this embodiment are the same as those in Example 1, except that the method of immunizing laying hens with the obtained main outer membrane protein immunostimulatory complex is changed to three immunizations, and the specific method is: the first immunization dose: 30μg-60μg / chicken, 15 days after the first immunization for the second immunization, immunization dose: 40μg-80μg / chicken; 15 days after the second immunization for the third immunization, immunization dose: 40μg-80μg / chickens, eggs were collected for 60 consecutive days after the 15th day after the third immunization. And the method for extracting egg yolk from egg is to get egg yolk after disinfecting egg shell with iodine.

Embodiment 3

[0077] The remaining steps of this embodiment are the same as those in Embodiment 2, except that the method for extracting the egg yolk is to use 10% double-chain polyamine chloride salt to sterilize the shell of the egg and then take the egg yolk.

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Abstract

Production of various Aeromonas hydrophila protective antigen monovalent and multivalent vitelline antibody and its usage in prevention of aquatic animal diseases are disclosed. It utilizes minor-protective antibody to make immune irritant composite egged hen and produce vitelline antibody product. It's safe, efficient, fast, has no harm and residue. It has more output, better immune-system development and functions of nutrients-added, disease-prevention and health-care.

Description

technical field [0001] The present invention relates to a kind of biological immunity technology, especially a preparation method of main protective antigen monovalent and multivalent egg yolk antibodies of Aeromonas hydrophila suitable for the prevention and treatment of passive immunity of aquatic animals, and their antiviral effect on aquatic animal diseases. application in prevention. technical background [0002] Yolk antibody (IgY), as a new class of drugs and feed nutritional additives, has been widely used in the prevention and treatment of bacterial or viral diseases in livestock such as pigs, cattle, and poultry such as chickens, ducks, and geese. Studies have proved that the purified egg yolk antibody and egg yolk liquid can be stored in freeze-dried or spray-dried storage without loss of antibody activity, acid resistance, heat resistance and stable performance. The polyvalent egg yolk antibody has no toxic and side effects, and long-term use will not lead to dr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02A61K39/395A61P31/04A23K1/18A23K1/16A23K20/195A23K50/80
Inventor 林天龙宋铁英俞伏松曾丽莉
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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