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A kind of egg yolk antibody with high biological activity and preparation method thereof

A kind of egg yolk antibody, biological activity technology

Inactive Publication Date: 2011-12-14
JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no public production technology for the yolk antibody against Aeromonas hydrophila, the main pathogen of fish fulminant hemorrhagic disease.

Method used

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  • A kind of egg yolk antibody with high biological activity and preparation method thereof
  • A kind of egg yolk antibody with high biological activity and preparation method thereof
  • A kind of egg yolk antibody with high biological activity and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The cultivation of embodiment 1 Aeromonas hydrophila F8 (provided by Shanghai Ocean University, isolated from grass carp)

[0025] Aeromonas hydrophila was inoculated on an agar plate or nutrient broth (pH7.0-7.5), cultured at 28-30°C, and the logarithmic growth phase was 6-18 hours after inoculation. The colony shape is smooth, the edges are neat, the surface is moist, round, slightly convex, with a special aroma, white to light yellow.

Embodiment 2

[0026] Example 2 Extraction of Aeromonas hydrophila aerolysin and preparation of oil seedlings

[0027] Aeromonas hydrophila was cultured with LB based on 28-30°C for 48-60h, centrifuged at 4500rpm to get the supernatant, first precipitated with 20% saturated ammonium sulfate at 4°C for 4h, centrifuged at 5000rpm, took the supernatant, and then used 60 Precipitate ammonium sulfate with % saturation, overnight at 4°C, centrifuge at 5000rpm, dissolve and dialyze with 50mmol / L Tris-HCl (pH=7.8~8.0) for two days, centrifuge at 12000rpm for 5min, remove impurities, and adjust the protein concentration of the supernatant to 100μg / mL, Freund's adjuvant equal to the supernatant is fully emulsified until it does not disperse within 1 min after being dropped into water.

Embodiment 3

[0028] The extraction of embodiment 3 extracellular protease ECPase and the preparation of oil seedling

[0029] Aeromonas hydrophila was cultured with LB based on 28-30°C for 48-60h, centrifuged at 4500rpm, supernatant was taken, precipitated with 70% saturated ammonium sulfate, overnight at 4°C, centrifuged at 5000rpm, washed with 50mM Tris-HCl (pH7 .8 to 8.0) were dissolved and dialyzed for two days, centrifuged at 12,000 rpm for 5 minutes to remove impurities, and the protein concentration of the supernatant was adjusted to 200 μg / mL, and fully emulsified with Freund's adjuvant in an amount equivalent to the supernatant.

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Abstract

The invention discloses a highly biologically active yolk antibody and a preparation method thereof. The antibody in the present invention uses aerolysin and extracellular protease, the main pathogenic factors of pathogenic Aeromonas hydrophila, as immunogens respectively, and simultaneously immunizes laying hens with an appropriate immunization program to obtain highly immune eggs, and the titer can be as high as 1:212, the antibody was purified by octanoic acid-ammonium sulfate method and then spray-dried. The antibody titer obtained according to the invention is high, and can be used to control fish bacterial diseases, especially the immune protection effect on fish with hemorrhagic diseases can be as high as 90%. Moreover, the mass production of antibodies by this method is low in cost, labor-saving and workshop-saving, high in social benefits, and broad in prospects.

Description

technical field [0001] A preparation technology of anti-aerolysin and extracellular protease composite enhanced egg yolk antibody dry powder, which is used for the prevention and treatment of fish bacterial diseases. The invention belongs to the technical field of biological products and industrialized separation. Background technique [0002] Aeromonas hydrophila (Aeromonas hydrophila) is a kind of common bacteria that widely exists in the water environment. It is the main pathogen of bacterial diseases such as fulminant hemorrhagic disease in freshwater cultured fish in my country. The diseases caused by it in the aquatic production process There are many types of hazards, a wide range of epidemics, and high mortality, which brings great losses to aquaculture. The traditional treatment is to use antibacterial drugs, sometimes combined with traditional Chinese medicine preparations. However, the use of antibacterial drugs not only brings about serious drug resistance probl...

Claims

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Application Information

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IPC IPC(8): C07K16/12C07K16/02C07K1/30A61K39/40A61P31/04
Inventor 朱善元王帅兵左伟勇陆辉邢晓玲孟婷郑义成大荣王安平王永娟吴双洪伟鸣杨海峰黄文强秦枫
Owner JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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