Effect of low culture temperature on specific productivity, transcription level, and heterogeneity of erythropoietin in chinese hamster ovary cells

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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 37degreesC. At a culture temperature lower than 37degreesC cell growth was suppressed, but cell viability remained high for a longer culture period. When the culture temperature was lowered from 37degreesC to 33degreesC, more than a 2.5-fold increase in the maximum EPO concentration was achieved. This enhanced EPO production at 33degreesC was not just because of the extended culture longevity with the decreased release of proteolytic enzymes from dead cells, but mainly because of enhanced q(EPO). The q(EPO) at 33degreesC was 0.35 +/- 0.08 mug/10(6) cells/h, which was approximately 4-fold higher than that at 37degreesC. Although the highest q(EPO) Of 0.49 +/- 0.14 mug/10(6) cells/h was obtained at 30degreesC, 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 33degreesC in regard to isoform pattern, sialic acid content, and in vivo biological activity was comparable to or even better than that produced at 37degreesC. Taken together, the results obtained demonstrate the potential of the application of low culture temperature to the commercial EPO production in rCHO cells. (C) 2003 Wiley Periodicals, Inc.
Publisher
JOHN WILEY & SONS INC
Issue Date
2003-05
Language
English
Article Type
Article
Keywords

MONOCLONAL-ANTIBODY PRODUCTION; BUTYRATE-INDUCED APOPTOSIS; ALPHA-AMIDATING ENZYME; SODIUM-BUTYRATE; CHO CELLS; MAMMALIAN-CELLS; RECOMBINANT ERYTHROPOIETIN; PERFUSION CULTURES; NUCLEOTIDE POOLS; MOUSE HYBRIDOMA

Citation

BIOTECHNOLOGY AND BIOENGINEERING, v.82, pp.289 - 298

ISSN
0006-3592
URI
http://hdl.handle.net/10203/79351
Appears in Collection
BS-Journal Papers(저널논문)
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