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Purification and inactivation method of yolk antibody

A technology of egg yolk antibody and inactivation, applied in the field of purification and inactivation of egg yolk antibody, can solve the problems of low purity of egg yolk antibody and weak cellular immunity, and achieve the effects of good purity, low preparation cost and simple process

Active Publication Date: 2017-07-25
河南后羿生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is simple to operate and low in preparation cost, but the purity of the obtained egg yolk antibody is also low, and the inactivated antibody can only induce humoral immunity in the body, and has little effect on cellular immunity

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0033] The method for purifying and inactivating the anti-gosling plague freeze-dried egg yolk antibody comprises the following steps:

[0034] 1) Take 100 gosling plague-free eggs, disinfect and fumigate the eggs, beat the eggs by hand to separate the egg yolks;

[0035] 2) Add 3 times the volume of sterilized PBS buffer solution with a concentration of 0.01mol / L and pH 7.2 to the egg yolk, stir well, add sterile polyethylene glycol 6000, and stir while adding, so that the final concentration is 12% , continue to stir for 30 minutes, centrifuge at 3500 rpm for 10 minutes, and take the precipitate;

[0036] 3) Restore the precipitate to the volume of the original egg yolk with PBS buffer (same as above), then add 6 times the volume of acidified water (hydrochloric acid solution) pre-cooled to 2°C and pH 4.0, stir well, let stand at 4°C for 16 hours, and extract the Serum;

[0037] 4) Heating the supernatant to 22°C, and adding n-octanoic acid with a final concentration of 0....

Embodiment 2

[0044] The purification and inactivation method of anti-porcine pseudorabies virus freeze-dried egg yolk antibody comprises the following steps:

[0045] 1) Take 100 porcine pseudorabies virus high-immunity eggs, disinfect and fumigate the eggs, beat the eggs by hand to separate the egg yolks;

[0046] 2) Add 2.5 times the volume of sterile PBS buffer solution with a concentration of 0.01mol / L and pH 7.2 to the egg yolk, stir well, add sterile polyethylene glycol 4000, and stir while adding, so that the final concentration is 15% , continue to stir for 30 minutes, centrifuge at 3000rpm for 15min, and take the precipitate;

[0047] 3) Restore the precipitate to the volume of the original egg yolk with PBS buffer (same as above), then add 5 times the volume of acidified water (hydrochloric acid solution) pre-cooled to 1°C and pH 4.2, stir well, let stand at 2°C for 20 hours, and extract the Serum;

[0048] 4) Heating the supernatant to 18°C, and adding n-octanoic acid with a f...

Embodiment 3

[0055] The method for purifying and inactivating the freeze-dried egg yolk antibody against Muscovy duck parvovirus comprises the following steps:

[0056] 1) Take 100 Muscovy duck parvovirus-free eggs, disinfect and fumigate the eggs, beat the eggs by hand to separate the yolks;

[0057] 2) Add 3.5 times the volume of sterile PBS buffer solution with a concentration of 0.01mol / L and pH 7.2 to the egg yolk, stir well, add sterile polyethylene glycol 8000, and stir while adding, so that the final concentration is 10% , continue to stir for 30 minutes, centrifuge at 4000rpm for 5min, and take the precipitate;

[0058] 3) Restore the precipitate to the volume of the original egg yolk with PBS buffer (same as above), then add 7 times the volume of acidified water (hydrochloric acid solution) pre-cooled to 4°C and pH 4.5, stir well, let stand at 8°C for 12 hours, and extract the Serum;

[0059] 4) Heating the supernatant to 18°C, and adding n-octanoic acid with a final concentrat...

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Abstract

The invention discloses a purification and inactivation method of a yolk antibody, and belongs to the technical field of biological products. The method comprises the following steps: 1) adding a buffer and polyethylene glycol in egg yolk until the final concentration of polyethylene glycol is 10%-15%, evenly mixing, centrifuging, and taking a precipitate; 2) adding a buffer and acidified water in the precipitate until the pH value is 4.2-5.0, evenly mixing, standing, and taking a supernatant; 3) adding n-caprylic acid into the supernatant until the final concentration is 0.1% to 0.3%, evenly mixing, standing, and taking a supernatant; 4) filtering and degerming the supernatant, and performing ultrafiltration and concentration to obtain a concentrated solution; and 5) using a thiophilic mixed mode adsorption technology for purification of the concentrated solution to obtain the purified egg yolk antibody. The preparation process is simple, the operation is convenient, the product has the advantages of high yield, good purity, long storage life, safe and reliable use, no residue and low preparation cost, and is suitable for industrial production and application.

Description

technical field [0001] The invention relates to a method for purifying and inactivating egg yolk antibodies, belonging to the technical field of biological products. Background technique [0002] Egg yolk antibodies refer to immunoglobulin (IgY) in egg yolk. The hen will transfer the immunoglobulin in the blood to the egg during cell division and laying eggs. For example, IgA and IgM in the blood will combine with other proteins in the oviduct to form the protein part of the egg, and IgG will transfer to the egg yolk through the egg yolk membrane internal. Because there are differences between egg yolk IgG and mammalian IgG in relative molecular mass, isoelectric point, and bond structure of Fc substrate on the cell membrane, it is specifically called IgY. [0003] IgY is a 7S immunoglobulin with a relative molecular mass of about 180 kD and contains two subunits, namely a heavy chain of 67-70 kD and a light chain of 22-30 kD. IgY in egg yolk has high purity and large con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02C07K1/34C07K16/08
CPCC07K16/02C07K16/081C07K16/085
Inventor 孙林杰徐进李厚伟
Owner 河南后羿生物工程股份有限公司
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