Study On The Purification Process Of MDCK Cell Matrix Influenza Vaccine

[Objective] Influenza causes huge damage to human health and economy every year,and getting a flu vaccine is an effective way to prevent influenza.The rapid and efficient production of vaccines in the short term is a challenge for manufacturers.Therefore,a cost-effective influenza vaccine production process is needed to support the huge demand for vaccines.Traditional influenza vaccines are mainly produced with the help of chicken embryos and have various limitations.For example,the supply of chicken embryos may be affected by the influenza pandemic.The cell-cultured influenza vaccine has gradually become a research and development hotspot in recent years,and some foreign companies have listed this vaccine.In this study,by comparing the purification process of different cell-cultured influenza vaccines and optimizing the purification process,a higher purity cell-cultured influenza vaccine stock solution was obtained.[Methods] After deep filtration,nuclease treatment and ultrafiltration,the BV69 influenza virus was subjected to one-step chromatography to optimize the purification conditions of Sepharose 4FF,Capto core 700 and Cellufine sulfate,and to inactivate the harvested virus and verify it‘s inactivation.The inactivated virus solution is subjected to two-step chromatography,including optimization of purification conditions for Capto Q and Q Sepharose xl.In view of considering the cost and efficiency of actual production,on the premise of ensuring the virus recovery rate,the two-step chromatography method combining Capto core 700 and Capto Q was initially selected as the purification option of BV69,H1N1,BY,H3N2.[Results] Taking the purification of BY virus as an example,after two steps of Capto core 700 and Capto Q chromatography,the final virus recovery was 61.5%,the hemagglutinin content was 45.4?g/mL,the total protein removal rate was 79.3%,and the total protein was 72.8?g/mL,the DNA removal rate was 99.3%,and the DNA content was 20pg/mL.The above indicators have met the requirements of Pharmacopoeia.Laid the foundation for large-scale production of influenza vaccine.[Conclusion] Through exploring a series of purification processes,the contaminated proteins and host DNA in the virus stock solution were removed,and finally all relevant indexes of the stock solution reached the proposed requirements.The purification process conditions for MDCK cell-cultured influenza vaccine established by this subject play an important role in the large-scale production of cellmatrix influenza vaccine.