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Process method of extracting egg yolk antibody of duck tembusu virus

A duck Tembusu virus and a technological method are applied in the field of egg yolk immunoglobulin preparation, can solve the problems of difficult removal, toxic reagents, low purity and the like, and achieve the effects of prolonging storage time, high recovery efficiency and reducing pollution

Inactive Publication Date: 2017-06-20
GUANGZHOU GREAT BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] All of the above methods have some shortcomings. Some reagents are toxic and difficult to remove in IgY solution; some IgY has low recovery rate and low purity, which affects the quality of the antibody.

Method used

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  • Process method of extracting egg yolk antibody of duck tembusu virus
  • Process method of extracting egg yolk antibody of duck tembusu virus

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A process for extracting egg yolk antibody against duck Tembusu virus disease, comprising the following steps:

[0036] (1) Collect high-immune eggs after immunization with inactivated duck Tembusu vaccine. After inspection and disinfection, egg whites and yolks are separated manually or mechanically. Add acidified water according to the volume ratio of egg yolk liquid and acidified water 1:6 (acidified water is obtained by adjusting the pH to 4.5 with hydrochloric acid), and lower the temperature to 2-8°C, then add egg yolk liquid, stir well, and set aside at 2-8°C. After 5 hours, the supernatant was separated. The supernatant was frozen at -18°C and then thawed at 4°C.

[0037] (2) Add 2% diatomaceous earth to carry out microfiltration, and adopt a 0.45um micromembrane during filtration.

[0038] (3) Add octanoic acid solution to the filtrate, so that the final concentration (v / v) of octanoic acid is 0.4%, stir well, let it stand for precipitation, and the water pha...

Embodiment 2

[0040] A process for extracting egg yolk antibody against duck Tembusu virus disease, comprising the following steps:

[0041] (1) Collect high-immune eggs after immunization with inactivated duck Tembusu vaccine. After inspection and disinfection, egg whites and yolks are separated manually or mechanically. Add acidified water according to the volume ratio of egg yolk liquid to acidified water (1:7) (acidified water is purified water and adjust the pH to 4.3 with hydrochloric acid), and cool down to 2-8°C, then add egg yolk liquid, stir well, and set aside at 2-8°C After 5 hours, the supernatant was separated. The supernatant was frozen at -18°C and then thawed at 4°C.

[0042] (2) Add 3% diatomaceous earth to carry out microfiltration, and adopt 0.45um micromembrane during filtration.

[0043] (3) Add octanoic acid solution to the filtrate so that the final concentration (v / v) of octanoic acid is 0.3%, stir well, let it stand for precipitation, and the water phase part is ...

Embodiment 3

[0045] A process for extracting egg yolk antibody against duck Tembusu virus disease, comprising the following steps:

[0046] (1) Collect high-immune eggs after immunization with inactivated duck Tembusu vaccine. After inspection and disinfection, egg whites and yolks are separated manually or mechanically. Add acidified water according to the volume ratio of egg yolk liquid to acidified water (1:5) (acidified water is purified water and adjust the pH to 4.7 with hydrochloric acid), and cool down to 2-8°C, then add egg yolk liquid, stir well, and set aside at 2-8°C After 5 hours, the supernatant was separated. The supernatant was frozen at -18°C and then thawed at 4°C.

[0047] (2) Add 2% diatomaceous earth to carry out microfiltration, and adopt a 0.45um micromembrane during filtration.

[0048](3) Add octanoic acid solution to the filtrate so that the final concentration (v / v) of octanoic acid is 0.8%, stir well, let it stand for precipitation, and the water phase part is...

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Abstract

The invention discloses a process method of extracting an egg yolk antibody of a duck tembusu virus. The process method comprises the following steps of: S1, directly mixing acidified water with a yolk original liquid; breaking an emulsifying state under the action of mechanical stirring and water, wherein in the pH environment, the solubleness of the egg yolk antibody is the minimum and non water soluble lipoproteins are precipitated, and oil and an aqueous solution are separated to form a precipitated; S2, adding a proper amount of diatomite into the aqueous solution extracted in the first step for micro-filtration, wherein in acidic condition, the content of proteins is almost while the settling amount of lipoproteins is relatively great; and S3, mixing the aqueous solution extracted in the step 2 with caprylic acid, wherein caprylic acid is reacted with impure proteins in the aqueous solution to form large-particle substances which are separated from an immune globulin aqueous solution to achieve a purpose of extracting the immune globulins. The method disclosed by the invention has the advantages of being high in antibody recovery rate, simple to operate and the like.

Description

technical field [0001] The invention belongs to the technical field of preparation of egg yolk immunoglobulin in biotechnology pharmacy, and in particular relates to a process for extracting egg yolk antibody of duck Tembusu virus disease. Background technique [0002] Chicken yolk immunoglobulin G antibody is a 7S immunoglobulin that is slightly different from mammalian IgG. The protein has a molecular weight of about 180kDa and contains two subunits, namely a heavy chain of 67-70kDa and a light chain of 22-30kDa, also known as IgY (Yolk Immunoglobulin), which exists in the yolk of the immunized hen. [0003] Experiments have shown that after immune stimulation, the hen produces an immune response. During the yolk maturation period in the oviduct, IgY in the blood can be selectively transferred to the yolk, and it is the only immunoglobulin in the yolk. As early as the 1960s, scientists discovered that IgY antibodies existed in chicken egg yolk, and its content was similar...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02
CPCC07K16/02C07K16/1081C07K2317/11
Inventor 魏平华王小庆
Owner GUANGZHOU GREAT BIOLOGICAL TECH
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