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Preparation method for duck viral hepatitis refine yolk antibody

A duck virus hepatitis, egg yolk antibody technology, applied in the direction of anti-virus immunoglobulin, egg-derived immunoglobulin, etc., can solve the problems of inconvenient storage, virus transmission, high storage cost, and achieve no toxic side effects, no chemical residues , a strong specific effect

Active Publication Date: 2009-12-23
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, my country mainly uses egg yolk antibody to prevent the occurrence of the disease. The egg yolk antibody produced by it has many disadvantages: 1. Egg yolk contains some pathogens that can be vertically transmitted to chicks, such as mycoplasma, salmonella, adenovirus and reovirus etc. If strict disinfection and inactivation are not carried out in the production of egg yolk antibodies, it can lead to artificial transmission of the virus
2. Bacteria commonly found on the surface of eggshells cannot be inactivated by crude methods; 3. High-concentration egg yolk liquid provides the most favorable conditions for the reproduction of contaminated bacteria; 4. Inconvenient storage and high storage costs; 5. Use Extremely inconvenient, because of its high viscosity, it is easy to block the syringe; 6. The stress response is relatively strong

Method used

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Examples

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Effect test

Embodiment 1

[0023] A kind of preparation method of duck viral hepatitis refined egg yolk antibody: its preparation method step is:

[0024] 1. Preparation of virus seed for production: the AV2111-5 strain virus seed that was purchased from the duck virus hepatitis seed virus AV2111 strain of the China Veterinary Drug Supervision Institute of the Ministry of Agriculture in the 10-day-old SPF chicken embryo for 5 generations was used to kill Bacterial saline for 10 -2 Dilute, inoculate well-developed 9-10-day-old SPF chicken embryos with the diluted virus seeds through the chorioallantoic cavity, incubate at 37°C, and harvest the allantoic fluid of chicken embryos that died within 48-96 hours and had typical embryo body lesions;

[0025] 2. Preparation of venom for antigen preparation: make 10 sterilized physiological saline with poisonous species for production -2 Dilute, inoculate 10-day-old chicken embryos into the chorioallantoic cavity, inoculate 0.1ml per embryo, and incubate at 37°C...

Embodiment 2

[0034] A preparation method of purified egg yolk antibody against duck viral hepatitis: the preparation method steps are: the preparation steps ①, ②, ③, ④, ⑤, ⑦, ⑧ are the same as the preparation steps ①, ②, ③, ④, ⑤ of Example 1 , ⑦, ⑧; steps ⑥, ⑨ become:

[0035] ⑥Inactivation I and extraction: Dilute the isolated egg yolk 1:1 with 0.015mol / L sterilized PBS pH7.2, heat inactivation at 60°C for 15 minutes; add ammonium sulfate with a final concentration of 16%, 8000 rpm / separation Incubate for 15 minutes and collect the pellet.

[0036] ⑨. Preparation and packaging: The collected sterile filtrate was subjected to duck liver neutralizing antibody titer determination, and the antibody was diluted with 0.015mol / L pH7.2 sterilized PBS to a neutralizing antibody titer of 1:512, and the final The concentration is 0.02% (volume percentage) Tween-80 as a stabilizer and fully mixed, subpackaged, and aseptically quantitatively subpackaged.

Embodiment 3

[0038] A preparation method of purified egg yolk antibody against duck viral hepatitis: the preparation method steps are: the preparation steps ①, ②, ③, ④, ⑤, ⑦, ⑧ are the same as the preparation steps ①, ②, ③, ④, ⑤ of Example 1 , ⑦, ⑧; steps ⑥, ⑨ become:

[0039] ⑥Inactivation I and extraction: Dilute the isolated egg yolk 1:1 with 0.015mol / L sterilized PBS pH7.2, heat inactivation at 60°C for 15 minutes; add ammonium sulfate with a final concentration of 19%, 8000 rpm / separation Incubate for 15 minutes and collect the pellet.

[0040] ⑨. Preparation and packaging: The collected sterile filtrate was subjected to duck liver neutralizing antibody titer determination, and the antibody was diluted with 0.015mol / L pH7.2 sterilized PBS to a neutralizing antibody titer of 1:1024, and the final The concentration is 0.02% (volume percentage) Tween-80 as a stabilizer and fully mixed, subpackaged, and aseptically quantitatively subpackaged.

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Abstract

The invention relates to a preparation method for a duck viral hepatitis refine yolk antibody, which comprises the following steps: (1) preparation of virus seed for production; (2) preparation of venom for preparing antigen; (3) preparation of oil-emulsion inactivated vaccine; (4) preparation of eggs from hype-immunized chickens; (5) separation of yolk; (6) inactivation I and extraction; (7) inactivation II, collation and deep filtration; (8) inactivation III and degerming filtration; and (9) mixing and package, wherein the collected sterile filtered liquid is subject to liver neutralizing antibody titer determination; the antibody is diluted until the neutralizing antibody titer of the antibody is 1:256-1:1,024 by using 0.015mol / L of sterilized PBS with the pH of 7.2; 0.02 volume percent of tween-80 as a stabilizer is added in the antibody, and the mixture is evenly mixed and sub-packaged; and the obtained product is sterile and quantitative sub-packaged. The product has the advantages of high purity, strong specificity, easy preservation, no residue and the like, and has extremely good effect of preventing and treating the duck viral hepatitis.

Description

Technical field: [0001] The invention relates to the technical field of poultry biopharmaceutical preparations, in particular to a method for preparing duck viral hepatitis refined egg yolk antibody. Background technique: [0002] Duck viral hepatitis (DVH) is an acute fatal infectious disease of ducklings. The disease has an acute onset, rapid spread, and morbidity and mortality rates can be as high as 100%. Since duck hepatitis virus (DHV) was first isolated in Shanghai in 1958 in my country, it has been prevalent in many provinces and cities across the country, causing huge economic losses, and has become an important threat to the healthy development of the duck industry. [0003] Duck viral hepatitis mainly threatens ducklings less than 20 days old. At this time, the immune system of ducklings is not yet fully developed, and it is difficult to achieve the preventive effect after vaccination. Although higher maternal antibodies can make ducklings immune within 2 weeks ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02C07K16/10
Inventor 孙进忠乔荣岑习向锋闵亚杰晋春霞
Owner PU LIKE BIO ENG
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