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Method for purification treatment of embryotoxin antigen and application of method

A technology for purifying and treating antigens, which is applied in the direction of virus antigen components, biochemical equipment and methods, viruses, etc., and can solve the problems of unguaranteed vaccine safety, great human harm, and the content of antigen-contaminated viruses

Active Publication Date: 2021-08-20
SHANDONG SINDER TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this production method also has some disadvantages. For example, because the antigen is harvested from the allantoic fluid of the poultry embryo, or even the embryo body, there must be many miscellaneous proteins and lipids in the poultry embryo itself. During the preparation of the vaccine, these impurities must be removed. , otherwise the safety of the vaccine cannot be guaranteed, and the stability of long-term storage is poor
Conventional treatment methods have some problems, such as simple centrifugal treatment can only remove impurities with large particles; chromatography treatment steps are complicated and costly; chloroform extraction is toxic and harmful to the human body, so Restricted use and not easy for large-scale production; repeated freezing and thawing methods are easy to contaminate the antigen and have a great impact on the virus content

Method used

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  • Method for purification treatment of embryotoxin antigen and application of method
  • Method for purification treatment of embryotoxin antigen and application of method
  • Method for purification treatment of embryotoxin antigen and application of method

Examples

Experimental program
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Effect test

Embodiment 1

[0024] The method for the purification treatment of embryotoxin antigens provided in this specific example is applied to duck viral hepatitis antigens produced by duck embryos in Cherry Valley, and the specific purification treatment method includes the following steps:

[0025] Harvest duck embryo allantoic fluid and embryo body, after fully grinding by colloid mill, use cup centrifuge at 7000rpm, centrifuge for 20min, take supernatant (this step is sampled and preserved, marked as sample A), supernatant is placed in advance preheating Put it into a water bath at 37°C, after the temperature of the supernatant rises to 37°C, add octanoic acid in an amount of 0.5% of the volume of the supernatant, stir while adding, and keep stirring for 5min, then place the stirring liquid at room temperature Leave it for 4 hours, then filter the upper layer of lipids with sterilized gauze. Add calcium chloride to the filtrate, the addition of calcium chloride is the mass percentage concentrat...

Embodiment 2

[0030] The method for the purification treatment of embryotoxin antigens provided in this specific example is applied to the production of gosling plague antigens in Dasanhua goose embryos. The specific purification treatment method includes the following steps:

[0031] After harvesting the goose embryo allantoic fluid, embryo body and allantoic membrane, use a colloid mill to fully grind it, then use a cup centrifuge at 6000rpm, centrifuge for 30min, take the supernatant (this step is sampled and preserved, marked as sample D), and the supernatant Place in a water bath preheated to 42°C in advance. After the temperature of the supernatant rises to 42°C, add octanoic acid in an amount of 0.5% of the volume of the supernatant, stir while adding, and keep stirring for 10 minutes, and then stir the liquid Stand at room temperature for 5h. Use sterilized gauze to filter the upper layer of lipids, and then add calcium chloride to the filtrate. The amount of calcium chloride added ...

Embodiment 3

[0036] The method for purification treatment of embryotoxin antigens provided in this specific example is applied to the production of goose paramycin antigens in chicken embryos, and the specific purification treatment method includes the following steps:

[0037] In this example, chicken embryos were used to inoculate goose paramyxovirus. After the allantoic fluid of chicken embryos was harvested, it was frozen and thawed once at -20°C, and centrifuged at 5000rpm for 15 minutes in a cup centrifuge, and the supernatant (sampling marked as sample G) was taken. The solids were collected and weighed to calculate a solids content of approximately 1.5%. Because only the allantoic fluid is collected for this batch of antigens, the lipid content in the antigens is very small, so the octanoic acid treatment step can be omitted. Add calcium chloride to the supernatant obtained by centrifugation, the addition of calcium chloride is the mass percentage concentration of calcium chloride ...

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Abstract

The invention discloses a method for purification treatment of an embryotoxin antigen and application thereof. The specific method for purification treatment of the embryotoxin antigen comprises the following steps: inoculating a poultry embryo with poison, mixing a harvested embryo body and an allantoic fluid, grinding and centrifuging the mixed embryo body and allantoic fluid, and enabling supernate to remain; adding octanoic acid into the obtained supernate, standing, and filtering to remove lipids; and adding calcium chloride and sodium polyacrylate into an obtained filtrate, standing and centrifuging to obtain supernate, namely antigen liquid. The treatment method provided by the invention is simple and easy to operate, is safe and environment-friendly, can thoroughly remove impure proteins in the antigen, and has no influence on the virus content.

Description

technical field [0001] The invention belongs to the technical field of antigen purification treatment, and in particular relates to a method for purification treatment of embryotoxin antigens and its application. Background technique [0002] Embryotoxin antigen is a kind of antigen obtained by the production of poultry embryo. The production of antigen by poultry embryo is a classic antigen production method. A considerable part of the vaccine production of livestock and poultry requires the production of antigen by poultry embryo, which is widely used. . However, this production method also has some disadvantages. For example, because the antigen is harvested from the allantoic fluid of the poultry embryo, or even the embryo body, there must be many miscellaneous proteins and lipids in the poultry embryo itself. During the preparation of the vaccine, these impurities must be removed. Otherwise, the safety of the vaccine cannot be guaranteed, and the stability of long-term...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02A61K39/00A61K39/125A61K39/23A61K39/155A61P31/14A61P1/16A61P31/20
CPCA61K39/12A61K2039/552A61P1/16A61P31/14A61P31/20C07K16/02C07K2317/11C12N2750/14334C12N2760/18034C12N2770/32434
Inventor 裴香玲王学波刘志亮罗济冠苑慕荣乔彦良张磊李朝阳马晓旭
Owner SHANDONG SINDER TECH