Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.ymben.2007.01.001
Title: Application of destabilizing sequences on selection marker for improved recombinant protein productivity in CHO-DG44
Authors: Ng, S.K.
Wang, D.I.C.
Yap, M.G.S. 
Keywords: AU-rich elements
CHO-DG44
Dihydrofolate reductase
Interferon-gamma
Mammalian cell culture
Methotrexate amplification
Murine ornithine decarboxylase
PEST region
Recombinant protein expression
Issue Date: May-2007
Source: Ng, S.K., Wang, D.I.C., Yap, M.G.S. (2007-05). Application of destabilizing sequences on selection marker for improved recombinant protein productivity in CHO-DG44. Metabolic Engineering 9 (3) : 304-316. ScholarBank@NUS Repository. https://doi.org/10.1016/j.ymben.2007.01.001
Abstract: We have developed a systematic and generic way to improve recombinant protein productivities in stable transfections by applying mRNA and protein destabilizing elements to reduce selection marker expression strength. Interferon-gamma (IFNγ) expression vectors containing different combinations of AU-rich elements (ARE) and mouse ornithine decarboxylase (MODC) PEST region on the amplifiable dihydrofolate reductase (dhfr) selection marker were stably transfected into CHO-DG44 cells. Improvements in specific IFNγ productivities were 1.7-, 6.6- and 13.3-fold with the application of ARE, MODC PEST, and both ARE and MODC PEST, respectively. To further enhance productivities, compatibility of the destabilizing sequences with methotrexate (MTX) amplification was validated by amplifying the transfected cells to 50 nM MTX. A 14- to 27-fold increase in specific IFNγ productivities were observed after amplification, indicating the compatibility of the two systems. A high specific IFNγ productivity of 1.05 pg/cell/day was also attained by the amplified cell pool with both ARE and MODC PEST. © 2007 Elsevier Inc. All rights reserved.
Source Title: Metabolic Engineering
URI: http://scholarbank.nus.edu.sg/handle/10635/63498
ISSN: 10967176
DOI: 10.1016/j.ymben.2007.01.001
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