Please use this identifier to cite or link to this item:
|Title:||Overexpression of cold-inducible RNA-binding protein increases interferon-gamma production in Chinese-hamster ovary cells.|
|Source:||Tan, H.K.,Lee, M.M.,Yap, M.G.,Wang, D.I. (2008-04). Overexpression of cold-inducible RNA-binding protein increases interferon-gamma production in Chinese-hamster ovary cells.. Biotechnology and applied biochemistry 49 (Pt 4) : 247-257. ScholarBank@NUS Repository. https://doi.org/10.1042/BA20070032|
|Abstract:||Culturing recombinant CHO (Chinese-hamster ovary) cells at low temperatures (30-33 degrees 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 degrees C, but not at 37 degrees C. In the present study we demonstrated that overexpression of CIRP at 37 degrees C can increase the recombinant-protein titre in CHO cells. Stable overexpression of CIRP at 37 degrees C improved the final titre of CHO IFN-gamma, a recombinant CHO cell line producing human IFN-gamma (interferon-gamma), by 25% in adherent culture and up to 40% in suspension culture. Real-time PCR analysis showed that the increase in the recombinant IFN-gamma titre could be attributed to increased recombinant IFN-gamma mRNA levels, while growth data showed that CIRP overexpression did not result in growth arrest in CHO IFN-gamma cells. Glycan analysis showed that the increase in IFN-gamma titre as a result of CIRP overexpression did not affect the site occupancy, glycan structures or sialic acid content of IFN-gamma. Using this strategy, the final IFN-gamma 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.|
|Source Title:||Biotechnology and applied biochemistry|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on Dec 12, 2017
checked on Dec 14, 2017
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.