Method for inactivating viruses during avian influenza H9 subtype vaccine production
A technology for the production of avian influenza viruses and vaccines, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of destroying the immunogenicity of pathogens and long time for formaldehyde inactivation
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Embodiment 1
[0018] Embodiment 1: Optimization of inactivation process conditions
[0019] Test material: H9N2 subtype avian influenza virus
[0020] Inactivation method: (1) Virus inoculation and cultivation: select LMH cells for vaccine preparation and subculture until the density of LMH cells reaches 1.0×10 6 -1.5×10 6 Each / mL was inoculated with avian influenza virus H9 subtype, and added to cell maintenance medium for propagation and culture; (2) Virus collection, concentration and purification: the cell culture medium in step (1) was frozen and thawed three times, and stored at 4°C, After centrifugation at 7000rpm for 15 minutes, collect the avian influenza virus H9 subtype supernatant, and ultrafilter and concentrate the above supernatants respectively to obtain the avian influenza virus H9 subtype virus venom; (3) Virus inactivation: in the negative pressure production area, According to the amount of virus liquid, add BPL with final concentrations of 0.005%, 0.015%, 0.025%, 0.05...
Embodiment 2
[0032] Embodiment 2: vaccine preparation
[0033] Vaccine preparation: Mix 94 parts of white oil, 6 parts of Tween, and 1.5 parts of aluminum stearate to prepare the oil phase, mix and emulsify the solution of avian influenza virus and the oil phase according to the volume ratio of 1:2, and prepare the H9 subtype of avian influenza vaccine.
[0034] Table 3: Vaccines prepared by different inactivation processes
[0035]
Embodiment 4
[0036] Embodiment 4: BPL inactivated vaccine safety test
[0037] Test method: 60 SPF chickens were randomly divided into 6 groups, 10 birds in each group, among which 5 batches of BPL inactivated vaccine were used for safety evaluation in groups 1-5, and 2.5 mL was injected intramuscularly into the chest of each chicken. 6 groups of blank control group, no inoculation.
[0038] Safety test results: 14 days after immunization, the chickens in each treatment group were healthy, and the vaccine was well absorbed by necropsy, and no abnormalities appeared.
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