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A kind of preparation method of egg yolk antibody of duck astrovirus

An astrovirus, egg yolk antibody technology, applied in the direction of anti-virus immunoglobulin, egg-derived immunoglobulin, etc., can solve the problem of economic loss in duck farming industry

Active Publication Date: 2021-06-15
广东渔跃生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no duck astrovirus vaccine on the market, and there is no related drug to control the disease caused by the virus. Its occurrence often causes heavy economic losses to the duck industry

Method used

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  • A kind of preparation method of egg yolk antibody of duck astrovirus

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preparation example Construction

[0016] (1) Preparation of passaged cell line LMH cells: LMH cells were digested with 0.025% EDTA-trypsin to disperse and passaged, and cultured in DMEM containing 5-10% newborn bovine serum and appropriate amount of double antibody at 37°C, 5% CO 2 Cultivated in an incubator;

[0017] (2) Seed virus propagation: inoculate the LMH cells prepared in step 1 with the duck astrovirus type I virus seed according to the final volume of 1:200, and absorb the virus solution at 37°C for 30 minutes, and then use the virus containing 1~ 2% newborn bovine serum, appropriate amount of double antibody and 2mM glutamine in DMEM medium at 37°C, 5% CO 2 Cultivate to 58±2 hours;

[0018] (3) Virus collection, concentration and purification: when LMH cells have 80% cytopathic changes, the harvested cell venom is repeatedly frozen and thawed twice at -20°C, centrifuged at 5000rpm, 4°C for 10min, and the supernatant, i.e. the virus stock solution, is collected. The titer of the clear virus soluti...

Embodiment 1

[0027] Preparation:

[0028] (1) Use duck astrovirus type I inactivated vaccine to immunize 100-day-old Hailan brown chickens to obtain high-immunity eggs, specifically: subcutaneously inject 1 mL duck astrovirus type I inactivated vaccine Live vaccine immunization, a total of 3 immunizations, the time interval between two adjacent immunizations is 21 days, and eggs are collected 5 days after the last immunization to obtain high-immunity eggs;

[0029] (2) Disinfect the surface of the high-free eggs with 75% alcohol, wash them with water, separate the egg yolks, pass through a 60-mesh nylon sieve to remove the vitelline membrane and frenulum egg yolks, and break them to obtain egg yolk liquid;

[0030] (3) The egg yolk liquid obtained in step (2) is mixed with purified water at a volume ratio of 1:1, and after stirring evenly, adding citric acid and 2% polyanion cellulose with a weight volume ratio of 0.8%, after fully stirring, Place at 5°C for 6h, centrifuge at 10,000r / min ...

Embodiment 2

[0033] Preparation:

[0034] (1) Use duck astrovirus type I inactivated vaccine to immunize 100-day-old Hailan brown chickens to obtain high-immunity eggs, specifically: subcutaneously inject 1 mL duck astrovirus type I inactivated vaccine Live vaccine immunization, a total of 3 immunizations, the time interval between two adjacent immunizations is 21 days, and eggs are collected 5 days after the last immunization to obtain high-immunity eggs;

[0035] (2) High-free eggs are surface-sterilized with 75% alcohol, and the yolk is separated, passed through a 60-mesh nylon sieve to remove the yolk membrane and frenulum, and then placed in a colloid mill to grind for 5 minutes to obtain egg yolk liquid;

[0036] (3) The egg yolk liquid obtained in step (2) is mixed with purified water at a volume ratio of 1:3, and after stirring evenly, adding citric acid and 4% polyanion cellulose with a weight volume ratio of 2.5%, after fully stirring, Place at 1°C for 6h, centrifuge at 11000r / m...

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Abstract

The invention discloses a preparation method of duck astrovirus type I yolk antibody, which comprises the following steps: (1) using duck astrovirus type I inactivated vaccine to immunize breeding-aged hens to obtain highly immune eggs; (2) collecting High-free egg yolk, remove the yolk membrane and frenulum, and use a colloid mill to crush it to obtain egg yolk; (3) Mix the egg yolk obtained in step (2) with purified water at a volume ratio of 1:1 to 3, and stir evenly Finally, add citric acid with a final concentration of 0.8-2.5% weight-volume ratio and 2-4% polyanionic cellulose, stir well, place at 1-5°C for 3-6 hours, centrifuge at 1-5°C, and collect the supernatant; (4) Filter and sterilize the supernatant collected in step (3) to obtain egg yolk antibody liquid. The preparation method of duck astrovirus type I yolk antibody disclosed by the invention has simple steps, high efficiency and yield, less addition, and is economical and applicable.

Description

technical field [0001] The invention relates to a preparation method of duck astrovirus type I egg yolk antibody. Background technique [0002] The mortality rate caused by duck astrovirus type I is related to the age of ducklings. If it infects ducks aged 1 to 2 weeks, the mortality rate can reach 50%. If the disease occurs in ducks aged 4 to 6 weeks, the mortality rate is 10 to 25%. %. It is also pathogenic to adult laying ducks, which can lead to a decline in egg production, and the resulting mortality rate generally does not exceed 30%. Therefore, the disease has become one of the major diseases that seriously affect the production performance of duck farms. At present, there is no duck astrovirus vaccine on the market, and there is no related drug to control the disease caused by the virus. Its occurrence often causes heavy economic losses to the duck industry. [0003] Yolk antibody (IgY) is the immunoglobulin in the blood of the mother poultry that is transferred t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/02
CPCC07K16/02C07K16/10
Inventor 张毓金谢秉超敖艳华严悌昆黄淑芬
Owner 广东渔跃生物技术有限公司
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