篮式生物反应器制备Vero细胞乙型脑炎灭活疫苗

目的建立篮式生物反应器制备Vero细胞乙型脑炎灭活疫苗的工艺。方法利用7.5L篮式生物反应器和片状载体培养Vero细胞,接种乙型脑炎病毒P3V2株毒种,根据葡萄糖的消耗量,分析细胞的生长情况及调节病毒培养时的灌流速度,每24h取样,检测病毒滴度。收获的病毒液经纯化后,制备乙脑灭活疫苗,检测各项指标。结果Vero细胞培养至96h,葡萄糖消耗量达高峰,细胞密度达峰值;接种病毒后72h,葡萄糖消耗量达高峰,灌流量为7L/d,连续收获7～9d,共可收获(40±5)L病毒液;96h病毒滴度达高峰,为10.0LgLD50/ml;制备的乙型脑炎灭活疫苗各项指标均达到《中国药典》三部(2005版)要求。结论已建立了篮式生物反应器制备Vero细胞乙型脑炎灭活疫苗的工艺。

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.