Method for concentration and purification of antigens of avian influenza (H5N1) or porcine reproductive and respiratory syndrome (PRRS) viral vaccines

A technology for respiratory syndrome and virus vaccines, applied in the field of antigen concentration and purification, agricultural biology, can solve the problems of low antigen concentration and high cost of antigen purification process, and achieve the effect of improving specific activity

Inactive Publication Date: 2017-03-29
广东永顺生物制药股份有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to overcome the shortcomings of low vaccine antigen concentration, containing a large amount of host protein, and high cost of antigen purification process, and applies centrifugation and filtration technology to th

Method used

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Examples

Experimental program
Comparison scheme
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Example Embodiment

[0026] Example 1: A method for antigen concentration and purification of porcine reproductive and respiratory syndrome (PRRS) virus vaccine, including the following steps:

[0027] Step 1: Virus cultivation and harvest

[0028] (1) Virus seed reproduction:

[0029] Take the well-growing MARC-145 cells, digest them with trypsin, inoculate them in the cell bottle, inoculate the super mutant porcine reproductive and respiratory syndrome virus at 1% of the inoculum, and culture them at 37℃. Observe the cytopathic changes daily. When more than% of the cells have CPE, collect the poison (about 4 days), freeze and thaw twice, subpackage and sample for identification;

[0030] (2) Cell seed propagation:

[0031] Take out the cell tube from the liquid nitrogen tank, place it in a 37°C water bath to thaw, transfer the cells into a centrifuge tube containing 10ml of serum-free medium, and centrifuge at 1000rpm for 5min. Suspend cells in DMEM medium with 10% FBS, 37°C, 5% CO 2 Cultivation, when t...

Example Embodiment

[0044] Example 2: A method for antigen concentration and purification of avian influenza (H5N1) virus vaccine, including the following steps:

[0045] Step 1: Virus cultivation and harvest

[0046] Take 9-day-old chicken embryos without immune maternal antibodies and inoculate H5N1 avian influenza virus via the allantoic cavity. After inoculation, incubate at 37°C for 36-72h to collect allantoic fluid;

[0047] Step 2: Inactivation of the virus:

[0048] Add formaldehyde to the virus solution at a ratio of 0.1% (V / V), mix well, incubate in a shaking incubator at 37°C for 8 hours, take it out, put it at room temperature for 16 hours, then transfer to 4°C for storage, and the shelf life is 7 days; or use 1 / 4000β-propiolactone is inactivated at 4°C for 12-36h, and then hydrolyzed at 37°C;

[0049] Step 3: Initial separation of virus liquid:

[0050] The chicken embryo allantoic fluid containing inactivated virus is filtered through a 0.65μm filter membrane to remove cell debris and micros...

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Abstract

The invention discloses a method for concentration and purification of antigens of avian influenza (H5N1) or porcine reproductive and respiratory syndrome (PRRS) viral vaccines. Through research on high efficiency, low cost and large-scale concentration and purification technologies of antigens of avian influenza (H5N1) and porcine reproductive and respiratory syndrome (PRRS) and comparison with the concentration and purification technologies, the optimal concentration and purification methods and routes aiming at different antigens are selected, the concentration multiple of the virus raw liquid, impurity protein content determination indexes and antigen purification recovery rate indexes are determined through repetitive study of the antigen concentration and purification processes and batch sample quality stability study, a large-scale concentration and purification technology and a production line suitable for large-scale antigen concentration and purification are built, and a safe and efficient vaccine product is developed. The method has the advantages of simple processes, low cost and large-scale production feasibility. The method builds an animal vaccine multi-virus antigen liquid concentration and purification technology, provides a key practical novel technology for production of animal biological products in China and greatly promotes the study and development of animal biological products in China.

Description

technical field [0001] The technology relates to the antigen concentration and purification of avian influenza (H5N1) or porcine reproductive and respiratory syndrome (PRRS) virus vaccines, and belongs to the field of agricultural biotechnology, specifically the field of antigen concentration and purification. Background technique [0002] Since Pasteur, the father of vaccines, successfully developed the attenuated anthrax, cholera and rabies vaccines, the vaccine preparation technology has made great progress, and the vaccine forms have also developed from inactivated vaccines, live attenuated vaccines to genetic engineering vaccines, polypeptide vaccines and nucleic acid vaccines. . As far as vaccines for livestock and poultry are concerned, the world is still dominated by conventional vaccines, and so is our country. Due to the long development cycle of recombinant vaccines, poor immune efficacy and high cost, conventional vaccines will still dominate the market for a lo...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61K39/12A61P31/14A61P31/16C12N7/00C12N7/02C12N7/06
Inventor 林旭埜林德锐张毓金齐冬梅唐海兵黄平容赵肃清朱思思
Owner 广东永顺生物制药股份有限公司
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