Effect of Low Culture Temperature on.pdfVIP

Effect of Low Culture Temperature on Specific Productivity, Transcription Level, and Heterogeneity of Erythropoietin in Chinese Hamster Ovary Cells Sung Kwan Yoon,1 Ji Yong Song,2 Gyun Min Lee1 1Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 371-1 Kusong-Dong, Yusong-Gu, Daejon 305-701, Korea; fax: 82-42-869-2610; e-mail: gmlee@mail.kaist.ac.kr 2Life Science RD, LG Chemical Investment, P.O. Box 61, Yusong-Gu, Science Town, Daejon 305-380, Korea Received 7 July 2002; accepted 1 October 2002 DOI: 10.1002/bit.10566 Abstract: To determine the effect of low culture tempera- peutically important proteins. In an effort to increase re- ture on erythropoietin (EPO) production in recombinant combinant protein productivity in CHO cells, the effects of Chinese hamster ovary (rCHO) cells, rCHO cells produc- ing EPO (LGE10-9-27) were cultivated at 30, 33, and 37°C. various environmental parameters such as pH (Borys et al., At a culture temperature lower than 37°C cell growth was 1993), pO2 (Lin et al., 1993), pCO2 (Kimura and Miller, suppressed, but cell viability remained high for a longer 1996), and osmolality (Ryu et al., 2000) on cell growth and culture period. When the culture temperature was low- recombinant protein production have been investigated ex- ered from 37°C to 33°C, more than a 2.5-fold increase in tensively. the maximum EPO concentration was achieved. This en- hanced EPO production at 33°C was not just because of Temperature is another key environmental parameter that the extended culture longevity with the decreased re- influences cell growth and recombinant protein production. lease of proteolytic enzymes from dead cells, but mainly Most mammalian cells, including CHO cells, are cultivated because of enhanced qEPO. The qEPO at 33°C was 0.35