摘要
目的通过离心试验,制备出一种高纯度的流行性乙型脑炎(乙脑)灭活纯化疫苗(非洲绿猴肾细胞)[Purified Inactivated Japanese Encephalitis Vaccine(Vero Cell),JEV]。方法生物反应器培养的Vero细胞乙脑病毒液,超滤浓缩、甲醛灭活后,应用硫酸鱼精蛋白处理,再经过高速离心,蔗糖密度梯度超速离心纯化,超滤除糖,配制成JEV(Vero细胞)。结果经检测,疫苗中Vero细胞脱氧核糖核酸残留量≤10pg(皮克,Picogram)/剂,蛋白含量≤1μg(微克)/剂,宿主蛋白残留量≤30ng(纳克,Nanogram)/ml,并获得较高的抗原回收率。结论经过优化硫酸鱼精蛋白处理和蔗糖密度梯度超速离心纯化,制得了高纯度的JEV(Vero细胞)。
Objective To prepare a purified inactivated Japanese encephalitis vaccine（JEV）by ultracentrifugation.Methods Japanese encephalitis supernatant was collected from bioreactor.After concentrated by ultra-filtration and inactivated by formaldehyde,the inactivated virus suspension was treated with protamine sulfate and purified by sucrose density gradient ultracentrifugation.The sucrose was removed by ultra-filtration.Results The residual of vero cells deaxyribonucleic acid（DNA）was reduced to 10pg/dose and total protein content was reduced to 1μg/dose,the host cell protein was decreased to 15ng/dose,and satisfied recovery was obtained.Conclusion A purified inactivated JEV was prepared by optimizing condition of protamine sulfate treatment and sucrose density gradient ultracentrifugation purification.
出处
《中国疫苗和免疫》
CAS
2011年第4期350-352,共3页
Chinese Journal of Vaccines and Immunization
