Development Of Purified Technics For Influenza Split Inactivate Vaccine

Influenza viruses have high infectiousness and can cause acute respiratory illness. Its epidemic feature is outbreak in short term and quickly spread. Influenza causes various degree epidemics including pandemic, part.epidemic and diverging, yet causes considerable morbility and mortality. Now there is no specific medicine to prevent anyone from getting the flu, but vaccination is regarded as the best method of influenza prevention.A complete technics of influenza split inactivate vaccine by the study of influenza viruses was established in the article. First, the virus-containing virus fluids were harvested, after Influenza viruses of H₁N₁), H₃N₂, B respectivly propagated in embryonated chicken eggs and incubated, Then the virus-containing fluids was concentrated, purified, split, the second purified and inactivated. Finally the three types of influenza virus-containing fluids were affiliated to vaccine. Furthermore the two purify metheods of sucrose density gradient centrifugation and Sepharose 4FF agarose chromatography were groped and compared and the optimization method considering industrialization was determined.The results of the quality analysis of the two different purified virus-containing fluids show that two different purified virus-containing fluids qualities all meet the specification of WHO and the two purified methods may all be used in preparation influenza vaccine. But in the discarding rate of mixed protein and ovalbumin, Sepharose 4FF agarose chromatography is better than sucrose density gradient centrifugation. The discarding rate of mixed protein of Sepharose 4FF agarose chromatography and sucrose density gradient centrifugation is respectivly 89.2% and 87.5%. The discarding rate of ovalbumin of Sepharose 4FF agarose chromatography and sucrose density gradient centrifugation is respectivly 99.3% and 97.5%. Howerver, sucrose density gradient centrifugation is better Sepharose 4FF agrose chromatography in reclaimation rate from purification. The reclaimation rate of sucrose density gradient centrifugation and Sepharose 4FF agrose chromatography is respectivly 40.7% and 37.5%. In Endotoxin, there is no difference for the two methods.As technological research findings will be industrialization, we choose the sucrose density gradient centrifugation as purified method and study the postorder work of purified virus-containing fluids. Finally the finished products of vaccine are detected and results show to meet the WHO specification.