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
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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 ...
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