应用篮式生物反应器制备Vero细胞人用狂犬病疫苗  

Preparation of Rabies Vaccine for Human Use with Vero Cells in Basket Bioreactor

作　　者：杨屹[1] 汝东宇[2] 郭秀侠[1] 崔文广[1] 苗丽[1] 刘岩[1] 王亚军[1] 蔡月红[1] 苑志刚[1] 

机构地区：[1]长春生物制品研究所疫苗研究室,长春130062 [2]吉林大学畜牧兽医学院兽医微生物与免疫学实验室,长春130062

出　　处：《中国生物制品学杂志》2011年第5期583-585,589,共4页Chinese Journal of Biologicals

摘　　要：目的应用篮式生物反应器制备Vero细胞人用狂犬病疫苗。方法以人用狂犬病疫苗株aGV为毒种,Vero细胞为培养基质,利用14 L生物反应器(500 g片状载体置于载体篮内)灌流式培养,连续收获的病毒液经超滤浓缩、灭活、柱层析纯化后,加入冻干保护剂,制备冻干人用狂犬病疫苗,检测冻干前后及37℃放置28 d后的疫苗效价。结果细胞密度最高可达1.0×107个/ml;病毒收获液毒力为7.5×106～3.4×108 FFU/ml;纯化后总蛋白质含量小于80μg/0.5 ml,Vero细胞蛋白质残留量≤3.5μg/0.5 ml,Vero细胞DNA残留量小于100 pg/0.5 ml,疫苗效价大于5.0 IU/0.5 ml。结论应用篮式生物反应器可制备高质量的人用狂犬病疫苗。Objective To prepare rabies vaccine for human use with Vero cells in basket bioreactor.Methods Rabies vaccine virus strain aGV was cultured in Vero cells in 14 L bioreactor（500 g of disk carriers in basket） by perfused culture.The continuously harvested virus liquid was concentrated by ultrafiltration,inactivated,and purified by column chromatography,then added with stabilizer and prepared into freeze-dried vaccine.The prepared vaccine was determined for titers before and after lyophilization and after storage at 37℃ for 28 d.Results The cell density reached 1.0 × 107 cells / ml at most,and the titer of harvested virus liquid was 7.5 × 106 ～ 3.4 × 108 FFU / ml.After purification,the total protein content,residual Vero cell protein content,residual Vero cell DNA content and vaccine titer were less than 80 μg / 0.5 ml,less than 3.5 μg / 0.5 ml,less than 100 pg / 0.5 ml and more than 5.0 IU / 0.5 ml.Conclusion Basket bioreactor may be used for preparation of rabies vaccine for human use with high quality.

关 键 词：生物反应器 片状载体 狂犬病疫苗 VERO细胞 

分 类 号：R373.9[医药卫生—病原生物学] R392-33[医药卫生—基础医学]