期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 105, 期 1, 页码 215-220出版社
WILEY
DOI: 10.1002/bit.22533
关键词
mRNA translation; cold-shock; CHOk1 cells; eIF2 alpha; recombinant protein; IRES
资金
- BBSRC [BB/D009375/1, BB/F018908/1]
- Biotechnology and Biological Sciences Research Council [BB/F018908/1, BB/D009375/1] Funding Source: researchfish
Chinese hamster ovary cells (CHO) are routinely used in industry to produce recombinant therapeutic proteins and a number of studies have reported increased recombinant mRNA levels at temperatures <37 degrees C. Surprisingly, the effect of reduced temperature on mRNA translation in CHO cells has not been investigated despite this process being highly responsive to environmental stresses. The relationship between low temperature culturing of CHO cells and mRNA translation was therefore investigated using labeling studies and dual luciferase reporter gene technology. Global protein synthetic capacity was not greatly affected at 32 degrees C but was diminished at lower temperatures. The expression of both cap-dependent and cap-independent (IRES driven) mRNA translated luciferase reporter gene activity was highest at 32 degrees C on a per cell basis and this was partially accounted for by increased mRNA levels. Importantly, post-translational events appear to proceed with higher fidelity and accuracy at 32 than 37 degrees C resulting in increased yield of active protein as opposed to an increase in total polypeptide synthesis. Therefore at 32 degrees C recombinant cap-dependent mRNA translation appears sufficient to maintain recombinant protein yields on a per cell basis and this is associated with processing. Biotechnol. Bioeng. 2010;105: 215-220. (C) 2009 Wiley Periodicals, Inc.
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