Effects of environmental factors on recombinant protein production in Chinese Hamster Ovary cells = CHO 세포에서 재조합 단백질의 생산에 미치는 환경 인자들의 영향

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For the optimization of recombinant protein production in Chinese hamster ovary (CHO) cells, the effects of various environmental factors such as temperature, pH on cell growth and protein quality were systematically investigated using CHO cell lines producing erythropoietin (EPO), humanized anti-4-1BB antibody, therapeutic antibody and follicle stimulating factor (FSH). To determine the effect of low culture temperature on erythropoietin (EPO) production in recombinant Chinese hamster ovary (rCHO) cells, rCHO cells producing EPO (LGE10-9-27) were cultivated at 30, 33, and 37℃, respectively. At a culture temperature lower than 37℃, cell growth was suppressed, but cell viability remained high for a longer culture period. When the culture temperature was lowered from 37℃ to 33℃, more than 2.5-fold increase in the maximum EPO concentration was achieved. This enhanced EPO production at 33℃ was not just because of the extended culture longevity with the decreased release of proteolytic enzymes from dead cells, but mainly because of enhanced specific EPO productivity ($q_{EPO}$). The $q_{EPO}$ at 33℃ was $0.35 ± 0.08 μg/10^6$ cells/hr, which was approximately 4-fold higher than that at $37℃. Although the highest $q_{EPO}$ of $0.49 ± 0.14 μg/10^6$ cells/hr was obtained at 30℃, the maximum EPO concentration was lowest because the detrimental effect of lowering culture temperature on cell growth outweighed its beneficial effect on $q_{EPO}$. Like $q_{EPO}$, the relative EPO mRNA content increased by lowering culture temperature, indicating that the increased transcription level of EPO was responsible in part for the enhanced $q_{EPO}$ at low culture temperature. The quality of EPO produced at 33℃ in regard to isoform pattern, sialic acid content and in vivo biological activity was comparable to or even better than that produced at 37℃. Taken together, the results obtained here demonstrate the potential of the application of the low culture temperature to the commerc...
Advisors
Lee, Gyun-Minresearcher이균민researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
2004
Identifier
240541/325007  / 020015850
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생명과학과, 2004.8, [ xiii, 166 p. ]

Keywords

ENVIRONMENTAL FACTORS; RECOMBINANT PROTEIN; CHO세포; 환경인자; CHO CELLS; 재조합 단백질

URI
http://hdl.handle.net/10203/27568
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=240541&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
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