【摘要】 研究了以自制胶原蛋白载体及C-DISK载体培养CHO细胞生产rHuEPO蛋白的过程,并以进口A-DISK载体作为对照培养。对培养过程中细胞贴壁过程、葡萄糖代谢速率、rHuEPO浓度及葡萄糖代谢速率与rHuEPO的浓度变化关系进行了实验研究分析和对比。结果表明:自制胶原蛋白载体培养细胞具有良好的生物代谢活力和蛋白表达能力,葡萄糖代谢速率高达14.2mmol?mL?1?h?1,比自制C-DISK载体高16.4%,比进口A-DISK载体高12.7%;在自制胶原蛋白载体上培养CHO细胞时EPO表达量也最高,培养10天后EPO浓度达到282U?mL?1,比自制C-DISK载体高30%,也比进口A-DISK载体高18%。自制胶原蛋白载体具有较佳的性价比,是一种高效、廉价的动物细胞培养载体。细胞的葡萄糖代谢水平对蛋白表达有着重要的影响,但两者又不同步,因此可用葡萄糖的代谢速率变化对该培养过程和目的蛋白表达进行适当监测和调控。更多还原

【Abstract】 The processes of culturing CHO(Chinese Hamster Ovary)cells to produce rHuEPO(recombinant human Erythropoietin)were investigated by using self-prepared collagen carrier, self-prepared C-DISK carrier and imported A-DISK carrier, respectively. The analysis and comparison of the anchorage of CHO cells, metabolic rates of glucose, expression concentration of rHuEPO and the relation between metabolic rate of glucose and expression concentration of rHuEPO for culturing CHO cells on three different carriers were experimentally carried out. The results show that self-prepared collagen carrier has the best bio-activity and highest protein expression in CHO cell culture. The maximum glucose metabolic rate of CHO cells cultured on self-prepared collagen carrier is as high as 14.2 mmol?mL?1?h?1,and which is 16.4% higher than that on C-DISK carrier, 12.8% higher than that on A-DISK carrier, the final rHuEPO expression concentration in CHO cell culture cultured on self-prepared collagen carriers reaches 282 U?mL?1 after culturing ten days, and it is 30% higher than that on C-DISK carrier, 18% higher than that on A-DISK carrier. The self-prepared collagen carrier is a kind of high efficient and cheaper carrier which can be used for large-scale animal cell culture. The experimental results also show that the metabolic rate of glucose and expression concentration of rHuEPO in CHO cell culture cultured on different carriers are strongly relevant, but they are not in synchronization. This means that CHO cell culture stage can be detected and regulated by measuring the change of glucose metabolic rate.更多还原