Optimization of Influenza B Vaccine Candidates Cultured in MDCK Suspension Cells
In order to produce recombinant influenza B vaccine candidate strain rIBV on a large scale,this study explored the optimal culture conditions of rIBV on MDCKs cells in serum-free medium,and then expanded to a bioreactor to optimize the process parameters.After 15 successive passsages,the genetic stability and viral titer of MDCKs cell derived viral strain rIBV were determined.By optimizing the virus culture conditions,we determined that the optimal culture conditions for rIBV proliferation on MDCKs cells were TOI of 72 hpi,MOI=0.001,CCI of 4.0 × 106 cells/mL,TPCK trypsin concentration of 2 μg/mL,50％DO and pH of 7.2±0.5.rIBV showed high genetic stability after 15 successive passages on MDCKs cells.Compared with the chicken embryo derived rIBV strain,the EID50 and TCID50 of rIBV strain were increased from 106 25/mL to 1095/mL and 108,25/mL respectively after 15 passages of adaptation to MDCKs cell culture.In this study,the culture process of recombinant influenza B vaccine candidate rIBV on MDCKs cells was optimized,and a MDCKs cell-derived vaccine candidate rIBV with high infectivity and high viral titer was obtained,which laid a foundation for downstream influenza vaccine production and purification process.
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