温度对微囊化重组CHO细胞生长和去氨普酶表达的影响

摘要

为了研究不同温度对微囊化rCHO细胞生长代谢和重组蛋白表达影响,采用不同温度培养微囊化rCHO细胞,经MTT法测定细胞生长曲线,生物传感器测定葡萄糖和乳酸代谢,纤溶平板测定去氨普酶(DSPA,Desmodus ro-tundus salivary plasminogen activator)蛋白表达量.结果表明,低温培养降低微囊化rCHO的增殖速率和细胞密度,但提高了重组蛋白的生产.可见温度显著影响微囊化rCHO的生长代谢和重组蛋白表达,33℃下可以将DSPA蛋白生产量最大提高41%.%To investigate the effects of temperature on microencapsulated recombinant CHO(rCHO)cells proliferation, metabolism and recombinant protein expression,microencapsulated rCHO cells were cultured under different temperature.MTT method was used to measure the cell density,biosensor was used to measure the metabolism of glucose and lactate,and fibrin plate assay was used to meas-ure the expression of DSPA protein.Lower temperature inhibited the cell growth rate and cell density,but improved the DSPA produc-tion capacity of microencapsulated rCHO.The DSPA production increased by 41%at 33℃compared with 37℃.