Preparation and verification of tetravalent influenza virus subunit vaccine

Objective To prepare the 4-valent influenza virus subunit vaccine. Methods Four strains of virus were inoculated into the allantoic cavity of healthy chicken embryo which were 9 to 11 days old. After incubated at 33-35 ℃ for 48-72 h, the embryos were cooled and the allantoic fluid was harvested. Following virus inactivation, lysis and purification, sterilization and filtration were applied to prepare a monovalent vaccine stock solution; polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity. The harvested four monovalent stocks are mixed to prepare a tetravalent influenza virus subunit vaccine. Finally, the finished product vaccine was fully verified according to the Pharmacopoeia of the People’s Republic of China (2015 edition). Results The tetravalent influenza virus subunit vaccine was successfully prepared, among which hemagglutinin accounted for 91.4% of the total protein. The hemagglutinin of the four vaccine strains were H1N1 for 30 µg/ml, H3N2 for 34 µg/ml, B1 for 36 µg/ml, and B2 for 30 µg/ml, respectively. Conclusion The four-valent influenza subunit vaccine produced meets the requirements of production and verification. Key words: Tetravalent influenza virus subunit vaccine; Influenza; Vaccine; Type B influenza virus