摘要
目的建立生物反应器培养分泌重组人干扰素β1a(IFNβ1a)的CHO细胞的工艺,探讨CHO细胞表达分泌IFNβ1a的反应动力学规律。方法应用15 L生物反应器悬浮微载体方式培养CHO工程细胞,比较不同时期细胞的生长形态、数量、生物活性、灌流量、葡萄糖消耗量及其他物理参数的变化规律。结果 15 L生物反应器中在初始pH 6.86～7.20,溶氧30%～70%,温度36.8～37.2℃,微载体4 g/L,罐流量6.5 L/h的条件下连续培养40 d,CHO工程细胞的密度维持在5×105个/ml之间,收获的细胞液中重组人IFNβ1a的生物活性为2.5×104～4.0×105IU/ml,葡萄糖消耗量在0.5～2.5 mg/ml之间。结论初步建立了15 L生物反应器培养分泌重组人IFNβ1a的CHO细胞的工艺,为进一步建立工业化生产工艺奠定了基础。
Objective To develop a procedure for culture of Chinese hamster ovary （CHO） cells secreting recombinant human interferon β1a （IFNβ1a） in bioreactor and investigate the kinetic regularity of secretion of IFNβ1a. Methods CHO cells were subjected to suspension culture on microcarriers in 15 L bioreactor, of which the growth, count, biological activity, perfusion rate, glucose consummation and other physical parameters were evaluated. Results The density of CHO cells reached 5 × 105 cells/ml after culture in 15 L bioreactor at initial pH value of 6. 86 ～ 7. 20, dissolved oxy- gen content of 30% ～ 70%, temperature of 36. 8 - 37.2 ℃, microcarrier content of 4 g/L and flow rate of 6. 5 L/h for 40 d, while the biological activity of recombinant human IFNβ1a in harvested cells was 2. 5 × 104 - 4. 0 × 105 IU/ml, and the glucose consumption was O. 5 ～ 2. 5 mg / ml. Conclusion A procedure for culture of CHO cells secreting recombi- nant human IFNβ1a in 15 L bioreactor was preliminarily developed, which laid a foundation of further development of large-seale oroduetion procedure of IFNβ1a.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第10期1493-1497,共5页
Chinese Journal of Biologicals
基金
广东省教育部产学研结合项目(2012B091100408)
深圳市科技创新委资助项目(2112K3070011
JAS2009032012-00A)
