Effect of simultaneous application of stressful culture conditions on specific productivity and heterogeneity of erythropoietin in Chinese hamster ovary cells.
A single stressful culture condition induced by hypoosmotic stress (210 mOsm kg(-1)), low culture temperature (32 degrees C), or NaBu addition (1 mM) resulted in a 1.8- to 2.2-fold enhancement of specific erythropoietin (EPO) productivity (qEPO) of recombinant Chinese hamster ovary (rCHO) cells compared to normal culture condition (37 degrees C and 310 mOsm kg(-1)). Simultaneous application of these stressful conditions further enhanced qEPO up to approximately 5-fold. However, the quality of EPO was affected by stressful culture conditions. The proportion of acidic isoforms of EPO under a single stressful condition was 2.8-13.8% lower than that under normal culture condition. Simultaneous application of the stressful conditions further decreased the portion of acidic isoforms but not significantly. Despite 5-fold enhancement of q(EPO), the portion of acidic isoforms under the simultaneous application of stressful culture conditions was 12.9-21.6% lower than that under normal culture condition. Taken together, these results suggest the potential of simultaneous application of different stressful culture conditions to the production phase of two-stage culture, where cell growth and production phases are separated, for improved EPO production.
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 371-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea.
Sung Kwan
SK
Yoon
Jong Kwang
JK
Hong
Gyun Min
GM
Lee
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004921:Erythropoietin / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D007525:Isoelectric Focusing; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D004921:Erythropoietin