Hydrophobia split vaccine for human body
A rabies virus and vaccine technology, applied in the field of virus split vaccine preparation, can solve the problems of inability to further increase antigen content, increase of vaccine side reactions, etc.
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Embodiment 1
[0033] Lysis, extraction and purification of rabies whole virus particles and vaccine preparation:
[0034] The Vero cells were cultured at 37°C and passed through four consecutive passages. After the 4th passage cells were cultured for 3 to 5 days, they were washed with phosphate buffer, and the virus titer inoculated was not less than 7.5LgLD 50 / ml of rabies virus fixed virus CTN-1V strain. Then add cell maintenance solution and incubate at 35°C for 24 hours. Replace with fresh cell maintenance solution, and continue culturing for 2 to 4 days according to the above conditions. When the cytopathic effect (CPE) reaches ++ to +++, harvest and combine the virus solution. Add β-propiolactone at a ratio of 1:4000, and place at 22°C for 8 days to inactivate the virus. After passing the inactivation test, it was concentrated by ultrafiltration with a 100KD filter membrane. 10 mg / ml of protamine sulfate was added to the concentrated virus liquid to make the final concentration 0....
Embodiment 2
[0036] Lysis, extraction and purification of rabies whole virus particles and vaccine preparation:
[0037] The Vero cells were cultured at 37°C and passed through four consecutive passages. After the 4th passage cells were cultured for 3 to 5 days, they were washed with phosphate buffer, and the virus titer inoculated was not less than 7.5LgLD 50 / ml of rabies virus fixed virus aGV strain. Then add cell maintenance solution and incubate at 35°C for 24 hours. Replace with fresh cell maintenance solution, and continue culturing for 2 to 4 days according to the above conditions. When the cytopathic effect (CPE) reaches ++ to +++, harvest and combine the virus solution. Add β-propiolactone at a ratio of 1:4000, and place at 22°C for 8 days to inactivate the virus. After passing the inactivation test, it was concentrated by ultrafiltration with a 100KD filter membrane. 10 mg / ml of protamine sulfate was added to the concentrated virus liquid to make the final concentration 0.1 ...
Embodiment 3
[0039] Safety test of rabies virus split vaccine:
[0040] The safety of the rabies virus split vaccine was assessed by abnormal toxicity tests in mice and guinea pigs. Choose clean-grade guinea pigs with a body weight of 250-350 g, use 2 guinea pigs for each vaccine specification, and inject 5.0 ml of rabies virus split vaccine intraperitoneally into each guinea pig, weigh them before and 7 days after immunization, and observe the vaccination reaction at any time. Choose clean-grade mice with a body weight of 18-22 g, use 5 mice for each vaccine specification, and inject 0.5 ml of rabies virus split vaccine into each mouse, weigh them before and 7 days after immunization, and observe the vaccination reaction at any time . The test results are listed in Table 1.
[0041]
[0042] Animal safety tests have shown that rabies virus split vaccines of different specifications prepared with different fixed strains of rabies are free from contamination by exogenous toxic s...
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