应用生物反应器无血清培养杂交瘤细胞生产治疗用WuT3单抗的工艺(pdf)

    【摘要】 目的 建立用生物反应器无血清大规模培养WuT3杂交瘤细胞生产体内治疗用单抗的工艺。方法 将WuT3细胞适应于无血清培养液中,并在方瓶及3.5L、14L、75L生物反应器中逐级放大培养。结果 细胞能适应无血清培养，培养规模能成功放大到75L规模，细胞浓度达1.5×106/ml,培养上清中单抗浓度>50μg/ml，达到含10%小牛血清(FCS)培养液培养的水平。在整个放大培养过程中细胞能稳定分泌单抗，细胞株支原体检测为阴性。收获的单抗活性及热原检测均合格。结论 建立了一套无血清培养杂交瘤细胞生产治疗用单抗的工艺，适于规模化生产。

    【关键词】 生物反应器；无血清培养液；杂交瘤细胞；单克隆抗体

    Technological procedure on no serum cultivation of WuT3 hybridoma cells in bioreactor for production of therapeutic mAb

    ZOU Chang-yong,ZHAN Qian,LENG Wu-jun,et al.Wuhan Institute of Biological Products,Wuhan 430060,China

    【Abstract】 Objective To establish the technological procedure on large-scale cultivation of WuT3 hybridoma cells in bioreactor for production of therapeutic mAb. Methods WuT3 hybridoma cells were cultured in SFM in flask and gradually scaled up to 3.5L,14L and 75L bioreactor. Results WuT3 hybridoma cells can be cultured in flask and 75L bioreactor.The cells density and mAb concentration in harvest supernatant were 1.5×106/ml and 60μg/ml,respectively,which were similar to those in 10%FCS.The cells were stable and were negative in mycoplasma test.The pyrogen test and activity of the harvest mAb were qualified. Conclusion The procedure on large-scale cultivation of WuT3 hybridoma cells in SFM for production of therapeutic mAb was successfully established. ......