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
View PDF5 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • 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 the preliminary separation of virus liquid, and then uses tangential ultrafiltration technology and chromatography technology to purify and refine the antigen , has the advantages of simple process, low cost, suitable for large-scale production, etc.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment one: a kind of antigen concentration and purification method of porcine reproductive and respiratory syndrome (PRRS) virus vaccine comprises the following steps:

[0027] Step 1: Virus Culture and Harvest

[0028] (1) Propagation of poisonous species:

[0029] Take well-grown MARC-145 cells, digest them with trypsin, inoculate them in cell flasks, inoculate them with 1% of the inoculum amount, and inoculate them with porcine reproductive and respiratory syndrome virus super mutant strains, culture them at 37°C, and observe the cell changes every day. When 70 When more than % of the cells appear CPE, the poison is collected (about 4 days), frozen and thawed twice, subpackaged, and sampled for identification;

[0030] (2) Cell seed propagation:

[0031] Take out the cell tube from the liquid nitrogen tank, put it in a 37°C water bath to melt, transfer the cells into a centrifuge tube containing 10ml of serum-free medium, and centrifuge at 1000rpm for 5min. S...

Embodiment 2

[0044] Embodiment two: a kind of antigen concentration and purification method of avian influenza (H5N1) virus vaccine comprises the following steps:

[0045] Step 1: Virus Culture and Harvest

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

[0047] Step 2: Inactivation of 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, place it at room temperature for 16 hours, and then transfer it to 4°C for storage. The storage period is 7 days; or use 1 / 4000β-propiolactone was inactivated at 4°C for 12-36h, then hydrolyzed at 37°C;

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

[0050] Chicken embryo allantoic fluid containing inactivated virus, filtered through a 0.65 μm filter membrane ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap