摘要
目的应用生物反应器培养细胞和病毒,大规模生产人用狂犬病疫苗。方法以巴斯德PV2061为毒种,以143代以内Vero细胞为培养基质,应用生物反应器,每升投放25g微载体,灌流式细胞培养,连续收获病毒液,经浓缩、灭活、纯化,制成Vero细胞狂犬病疫苗。结果细胞培养密度达1.2×107～1.5×107个/ml,病毒感染后可连续收获18～22d,病毒最高滴度8.5LogLD50/ml,平均滴度7.6LogLD50/ml。经柱层析纯化,杂蛋白去除率达99.95%以上,总蛋白含量≤80μg/g,DNA含量≤10pg/0.5ml,GP含量3.5～4.5IU/0.5ml,效力≥4.5IU/0.5ml。结论应用生物反应器细胞培养,可以大规模生产优质Vero细胞人用狂犬病疫苗。
Objective To prepare rabies vaccine for human use in a large scale by cell culture in bioreactor. Methods Prepare rabies vaccine by perfused culture of Vero cells within 143 passages inoculated with PV2061 strain in bioreactor containing 25 g/L of microcarrier. The virus bulk was continuously harvested,then concentrated,lnactivated and purified. Results The density of cultured cells reached 1.2×10^7 - 1.5×10^7 cells/ml. The virus bulk could be continuously harvested for 18 -22 d after inoculation and reached a highest titer of 8.50 LogLD50/ml and a mean titer of 7.2-7.6 LogLD50/mL After purification by column chromatography, the removal rate of foreign protein reached more than 99.95%. The total protein, DNA and GP contents of final bulk were not more than 80μg, not more than 10 pg and 3.5-4. 5 IU respectively,and its potency was not less than 4.5 IU/0. 5 ml. Conclusion The cell culture in bioreactor could be used for the large-scale production of rabies vaccine for human use.
出处
《中国生物制品学杂志》
CAS
CSCD
2006年第3期288-290,共3页
Chinese Journal of Biologicals
基金
辽宁省海外学子创业资金资助项目(辽科发[2005]40号).
