Preparation Process Of Subunit Influenza Vaccine

Influenza is severe respiratory disease caused by influenza virus which is highly infectious. At present, influenza split vaccine has a protective efficiency, but it can't efficiently protect under three years old children and the over sixty years old population from influenza infection. In order to solve this problem,influenza subunit vaccine with more economy, efficiency, and safety was developed in this thesis.Several important operations in the preparation process of influenza subunit vaccine were studied. First, the seed banks of three influenza virus strains H1N1,H3N2 and B were prepared under different culture conditions with various passage numbers. The results showed that these three influenza virus strains were stable in chick embryos less than five passages and theirmain seed banks and work seed banks were established. Besides, the effects of incubation temperature, harvest time and vaccinated chicken embryo age on influenza virustiter of coagulation were tested and the incubation processes were established, vaccination in the age of 11 days of chicken embryos at 35 ? and the harvest after 48 hours for H1N1 and H3N2,vaccination in the age of 10 days of chicken embryos at 35?, and harvest after 72 hours for strain B. The conditions for Inactivation of the virus were tested and inactivation operation was developed. H1N1 vaccine was inactivated with 0.1%formaldehyde for 72 h at 4?, while H3N2 and B vaccines with 0.05% formaldehyde at 4 ? for 48 h. Ultra filtration membranes with diffierent cutoffs and areas were tested for condensation. The purification operation was filted with 300 k and 2.5 m2 ultra filtration membrane and condensed ten times. The optimal split operation was conducted by adding 1.2% Triton N-101 into vaccine at 15~25? for 30~50min. The stability of the subunit influenza vaccine was tested under different conditions,indicating that the production process enabled the the vaccine efficiency.This research supported for the development and basic study of influenza virus vaccine.was helpful for the further development and industrial production of subunit influenza virus vaccine.