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https://scholarbank.nus.edu.sg/handle/10635/64365
DC Field | Value | |
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dc.title | Overexpression of cold-inducible RNA-binding protein increases interferon-γ production in Chinese-hamster ovary cells | |
dc.contributor.author | Hong, K.T. | |
dc.contributor.author | May, M.L. | |
dc.contributor.author | Yap, M.G.S. | |
dc.contributor.author | Wang, D.I.C. | |
dc.date.accessioned | 2014-06-17T07:46:22Z | |
dc.date.available | 2014-06-17T07:46:22Z | |
dc.date.issued | 2008-04 | |
dc.identifier.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. | |
dc.identifier.issn | 08854513 | |
dc.identifier.uri | http://scholarbank.nus.edu.sg/handle/10635/64365 | |
dc.description.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. | |
dc.description.uri | http://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1042/BA20070032 | |
dc.source | Scopus | |
dc.subject | Chinese-hamster ovary cells (CHO cells) | |
dc.subject | Cold-inducible RNA-binding protein (CIRP) | |
dc.subject | Fed-batch bioreactor | |
dc.subject | Glycosylation | |
dc.subject | Interferon-γ (IFN-γ) | |
dc.subject | Recombinant-protein productivity | |
dc.type | Article | |
dc.contributor.department | CHEMICAL & BIOMOLECULAR ENGINEERING | |
dc.description.sourcetitle | Biotechnology and Applied Biochemistry | |
dc.description.volume | 49 | |
dc.description.issue | 3-4 | |
dc.description.page | 247-257 | |
dc.description.coden | BABIE | |
dc.identifier.isiut | 000255385600008 | |
Appears in Collections: | Staff Publications |
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