Preparation of Inactivated Japanese Encephalitis Vaccine with Vero Cells Cultured on Fibra Disk in Cellingen Plus

Objective To develop a procedure for preparation of inactivated Japanese encephalitis(JE)vaccine with Vero cells cultured on Fibra disk in Cellingen Plus.Methods JE virus P3V2 seeds were inoculated to Vero cells cultured on Fibra disk in 7.5 L Cellingen Plus.According to the consumption of glucose,the cell growth was analyzed,and the perfusion rate for virus cul-ture was regulated.Samples were taken every 24 h for virus titration.The harvested virus liquid was purified to prepare inactivated JE vaccine.Overall control tests were performed on the prepared vaccine.Results Both glucose consumption and density of Vero cells reached peak values 96 h after culture.Seventy-two hours after inoculation of virus,glucose consumption reached a peak value,while the perfusion rate was 7 L/d.Virus liquid was harvested for 7～9 d and reached a total volume of(40±5)L.The virus titer reached a peak value of 10.0 LgLD50/ml 96 h after inoculation.All the quality indexes of prepared inactivated JE vaccine met the requirements in Chinese Pharmacopoeia(VolumeⅢ,2005 edition).Conclusion A procedure for preparation of inactivated JE vac-cine with Vero cells cultured on Fibra disk in Cellingen Plus was developed.