Split encephalitis B virus vaccine and method for preparing the same
A technology of Japanese encephalitis virus and Japanese encephalitis, applied in the field of preparation of virus split vaccine
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Embodiment 1
[0037] Lysis, Extraction and Purification of Japanese Encephalitis Virus Particles and Preparation of Vaccine
[0038] Type SA 14 ,P 3 strain. The Vero cells were cultured at 37°C and passed through four consecutive passages. After the 4th passage cells are cultured for 3-5 days, they are rinsed with phosphate buffer, and the inoculation virus titer is not less than 7.5LgLD 50 / ml Japanese encephalitis virus, and added to the cell maintenance solution at 35 ℃, cultured for 24 hours. Replace with fresh cell maintenance solution, and continue culturing for 2-4 days according to the above conditions. When the cytopathic pathology (CPE) reaches ++~+++, harvest and combine the virus solution. After the sample was reserved, 20% formaldehyde was added to make the final concentration 0.05%, and the virus was inactivated at 22°C for 8 days. After passing the inactivation test, it was concentrated by ultrafiltration with a 100KD filter membrane. 10 mg / ml protamine sulfate was adde...
Embodiment 2
[0040] Lysis, Extraction and Purification of Japanese Encephalitis Virus Particles and Preparation of Vaccine
[0041] Type SA 14 ,P 3 strain. The Vero cells were cultured at 37°C and passed through four consecutive passages. After the 4th passage cells are cultured for 3-5 days, they are rinsed with phosphate buffer, and the inoculation virus titer is not less than 7.5LgLD 50 / ml Japanese encephalitis virus, and added to the cell maintenance solution at 35 ℃, cultured for 24 hours. Replace with fresh cell maintenance solution, and continue culturing for 24 days according to the above conditions. When the cytopathic pathology (CPE) reaches ++~+++, harvest and combine the virus solution. After the sample was reserved, 20% formaldehyde was added to make the final concentration 0.05%, and the virus was inactivated at 22°C for 8 days. After passing the inactivation test, it was concentrated by ultrafiltration with a 300KD filter membrane. Add 10 mg / ml protamine sulfate to th...
Embodiment 3
[0043] Preparation of Japanese Encephalitis Virus Split Vaccine Absorbed by Aluminum Adjuvant
[0044] According to the virus protein content of Japanese encephalitis virus split vaccine stock solution, add sterilized non-pyrogenic aluminum hydroxide adjuvant to the vaccine stock solution. Add slowly and mix well to form a uniformly distributed suspension, pH 7.0-8.0. Each dose contains 70 μg of viral protein, aluminum hydroxide ≤ 0.70 mg, 0.5 ml per dose, and the vaccine is divided into packages.
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