生物反应器规模化培养重组CHO-C28细胞表达HBsAg的研究

应用新型聚酯纤维盘片,采用连续灌注培养方式,分别试验了细胞接种量、pH、DO、罐流速度等因素对CHO-C28细胞生长分泌HBsAg的影响,初步建立了5L生物反应器生产重组乙型肝炎疫苗的生产工艺。经3次试验培养,每次培养60d,较适宜的培养条件确定为:pH6.80-7.10,DO 20%-30%,温度36-37℃,灌流速度138ml/h,接种浓度1.9×106cell/ml。收获液的HBsAg平均滴度是1∶256,最高滴度可达1∶512,纯化后的HBsAg产率为0.912mg/L。最后对反应器培养工艺与现行的转瓶培养工艺进行了比较,生物反应器培养具有可控制培养条件、不易污染和可使HBsAg产率提高等优点。

Evaluation of a new microcarrier process in bioreactor in large-scale cultivation for hepatitis B vaccine

Bioreactor process for recombinant CHO-C28 cell cultivation of hepatitis B vaccine was investigated in this study.A piece of three dimensional polyester fiber discs was used as support material and a continuous perfusion mode was used for cultivation.The culture conditions such as inoculation of cell density,pH,DO and perfusion velocity were studied respectively.The continuous three run cultivations were performed,each run maintained for 60 days.The optimal culture conditions are in following pH 6.80~7.10,DO 20%~30%,temperature 36~37℃,perfusion velocity 138ml/h and inoculation cell density 1.9 106 cell/ml.The ultimate average titer for harvested culture is 1∶256 in RPHA,and the content of purified HBsAg is 0.912mg/L.Finally,the advantages and disadvantages for both culture systems of roller bottle and bioreactor were compared and summarized.