Preparation method of refined egg yolk antibody for duck viral hepatitis

A technology for duck viral hepatitis and egg yolk antibodies, which is applied to antiviral immunoglobulins and immunoglobulins from eggs, can solve the problems of non-specific antibody waste and selection of specific antibodies, so as to reduce the cost of antibody use and avoid wasteful effect

Active Publication Date: 2018-05-01
SHANDONG SINDER TECH
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Problems solved by technology

According to a large number of literature reports and laboratory test results, it is shown that the occurrence of duck viral hepatitis that is prevalent in my country is mainly caused by DHAV-1 and DHAV-3 infections, and the prevalence of a single serotype accounts for more than 70%. The mixed infection of the two serotypes accounts for less than 30%. However, the existing clinically available purified egg yolk antibodies to duck viral hepatitis are mainly bivalent antibodies, and no specific antibodies are selected for the serotypes prevailing in the local area.
There is almost no cross-protection between DHAV-1 and DHAV-3, which undoubtedly causes a waste of non-specific antibodies

Method used

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  • Preparation method of refined egg yolk antibody for duck viral hepatitis
  • Preparation method of refined egg yolk antibody for duck viral hepatitis
  • Preparation method of refined egg yolk antibody for duck viral hepatitis

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preparation example Construction

[0017] The preparation method of refined egg yolk antibody against duck viral hepatitis comprises the following steps: isolating duck viral hepatitis strain and preparing an inactivated vaccine; the inactivated vaccine is immunized with healthy laying hens to obtain highly immune eggs, and the egg yolk is acidified after being separated , caprylic acid treatment, centrifugation, plate and frame filtration, inactivation, ultrafiltration, liquid preparation, and filter sterilization to obtain the refined egg yolk antibody against duck viral hepatitis.

[0018] Among them, the following strains are preserved in China Center for Type Culture Collection:

[0019] Name Duck hepatitis virus SD-1 strain;

[0020] Deposit number CCTCC V201440;

[0021] The deposit date is November 2, 2014;

[0022] Referred to as duck hepatitis virus SD-1 strain;

[0023] Preservation address China. Wuhan. Wuhan University.

[0024] Name Duck hepatitis virus SD-3 strain;

[0025] Deposit number CC...

Embodiment 1

[0068] Example 1 Preparation of duck viral hepatitis DHAV-1 type and DHAV-3 type bivalent refined egg yolk antibody

[0069] 1) Separation of egg yolks: Take 250kg of DHAV-1 and DHAV-3 high-free eggs that have passed the above tests, totaling 500kg. After the eggshells are thoroughly sterilized with chloroform, the egg yolks are separated, and the egg yolk liquid obtained by separating the egg yolks is about It is 125kg.

[0070] 2) Acidification treatment: Prepare acetic acid-acetate buffer solution in proportion, weigh 375L water for injection, weigh 0.375kg anhydrous sodium acetate and 1.125 liters of glacial acetic acid in proportion, add to water for injection after weighing, and stir until completely dissolve. After dissolving, add ground egg yolk liquid whose weight ratio is 1 / 3 of the quality of the above-mentioned water for injection, and stir well for 30 minutes.

[0071] 3) Caprylic acid treatment: Slowly add 20 L of caprylic acid in proportion to the egg yolk liq...

Embodiment 2

[0092] Example 2 Preparation of duck viral hepatitis DHAV-1 type refined egg yolk antibody

[0093] 1) Egg yolk separation: Take 250 kg of DHAV-1 type high-free eggs that have passed the above tests, and thoroughly disinfect the eggshells with Neogilamine, then separate the egg yolks to obtain about 63 kg of egg yolk liquid.

[0094] 2) Acidification treatment: prepare acetic acid-acetate buffer solution in proportion, weigh 189L water for injection (3 times the weight of egg yolk liquid), weigh 0.189kg of anhydrous sodium acetate and 0.567 liter of glacial acetic acid in proportion, add injection after weighing In water, stir well until completely dissolved. After dissolving, add the ground egg yolk liquid and stir well for 30 minutes.

[0095] 3) Caprylic acid treatment: Slowly add 10 L of caprylic acid to the egg yolk liquid in proportion, and stir thoroughly for 60 minutes.

[0096] 4) Centrifugation: transfer the octanoic acid-treated material to a tubular continuous ce...

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Abstract

The invention provides a method for preparing duck viral hepatitis purified egg yolk antibody, comprising the following steps: isolating duck viral hepatitis strains and preparing an inactivated vaccine; immunizing disease-free laying hens with the inactivated vaccine to obtain highly immune eggs, and separating egg yolks After acidification treatment, octanoic acid treatment, centrifugation, plate and frame filtration, inactivation, ultrafiltration, liquid preparation and filter sterilization, the refined egg yolk antibody of duck viral hepatitis was obtained. Duck viral hepatitis type 1 and duck viral hepatitis type 3 strains were separately vaccinated, and healthy laying hens were immunized separately, avoiding the factors affecting the immune response competition of the two antigens in the same body, each serotype The antibody titer is controlled separately to avoid the consequences of immune tolerance to another serotype antigen caused by reimmunization with the double vaccine when the antibody titer of one serotype is not high. This technology has obtained high-titer refined egg yolk antibodies without lipid substances and any foreign microbial contamination.

Description

technical field [0001] The invention relates to the field of poultry biological products, in particular to a method for preparing duck viral hepatitis purified egg yolk antibody. Background technique [0002] Duck viral hepatitis is a highly fatal and rapidly spreading viral disease of ducklings caused by duck hepatitis virus (DHV), with hepatitis as its main feature. If the control is not proper, the mortality rate is very high, which will cause great economic losses to the duck farm. [0003] Duck viral hepatitis originated in Shanghai, my country in 1958. So far, the disease has been prevalent in my country for more than 50 years. According to a large number of literature reports and laboratory test results, it is shown that the occurrence of duck viral hepatitis that is prevalent in my country is mainly caused by DHAV-1 and DHAV-3 infections, and the prevalence of a single serotype accounts for more than 70%. Mixed infection of the two serotypes accounts for less than 3...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/02
Inventor 李朝阳赵锋祥乔彦良吴庆海薛希娟
Owner SHANDONG SINDER TECH
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