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Title: Overexpression of cold-inducible RNA-binding protein increases interferon-γ production in Chinese-hamster ovary cells
Authors: Hong, K.T.
May, M.L.
Yap, M.G.S. 
Wang, D.I.C.
Keywords: Chinese-hamster ovary cells (CHO cells)
Cold-inducible RNA-binding protein (CIRP)
Fed-batch bioreactor
Interferon-γ (IFN-γ)
Recombinant-protein productivity
Issue Date: Apr-2008
Citation: Hong, K.T., May, M.L., Yap, M.G.S., Wang, D.I.C. (2008-04). Overexpression of cold-inducible RNA-binding protein increases interferon-γ production in Chinese-hamster ovary cells. Biotechnology and Applied Biochemistry 49 (3-4) : 247-257. ScholarBank@NUS Repository.
Abstract: Culturing recombinant CHO (Chinese-hamster ovary) cells at low temperatures (30-33°C) increases specific recombinant protein productivity by 2-5-fold. However, even though the specific productivity is increased, cell growth is decreased in low-temperature culture such that the final recombinant protein titre remains unchanged or is even diminished, owing to the lower cell density. Exposing mammalian cells to low temperatures results in a change in the expression of many 'cold-stress' genes. CIRP (cold-inducible RNA-binding protein) is a cold-stress protein that is highly expressed at 32°C, but not at 37°C. In the present study we demonstrated that overexpression of CIRP at 37°C can increase the recombinant-protein titre in CHO cells. Stable overexpression of CIRP at 37°C improved the final titre of CHO IFN-γ, a recombinant CHO cell line producing human IFN-γ (interferon-γ), by 25% in adherent culture and up to 40% in suspension culture. Real-time PCR analysis showed that the increase in the recombinant IFN-γ titre could be attributed to increased recombinant IFN-γ mRNA levels, while growth data showed that CIRP overexpression did not result in growth arrest in CHO IFN-γ cells. Glycan analysis showed that the increase in IFN-γ titre as a result of CIRP overexpression did not affect the site occupancy, glycan structures or sialic acid content of IFN-γ. Using this strategy, the final IFN-γ titre was increased by 40% compared with current temperature-based strategies. Furthermore, there is no decrease in cell growth or recombinant-protein glycosylation quality, as previously observed in low-temperature culture. © 2008 Portland Press Ltd.
Source Title: Biotechnology and Applied Biochemistry
ISSN: 08854513
Appears in Collections:Staff Publications

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