Gadd45-induced cell cycle G2/M arrest for improved transient gene expression in Chinese hamster ovary cells

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Gadd45 is a p53-regulated protein and is involved in cell cycle arrest in the G2/M phase. In an effort to improve transient gene expression (TGE) in Chinese hamster ovary (CHO) cells, the effect of Gadd45-induced cell cycle arrest on TGE in CHO cells was investigated using the two different expression vectors encoding Fcfusion protein and recombinant antibody. To regulate the expression of Gadd45 in CHO cells, the CHO-TREx-gadd45 cell line was established using the T-REx system controlled by doxycycline. During the cultures for TGE, Gadd45 overexpression severely inhibited cell growth, but significantly enhanced TGE. Compared with the culture without Gadd45 overexpression, the TGE of Fc fusion protein and humanized antibody were increased by 111 and 93%, respectively. The enhanced TGE, despite the cell growth arrest induced by Gadd45 overexpression, was due to the significantly increased specific productivity, resulting from enhanced transfection efficiency, increased cell size, and active DNA demethylation. Taken together, the data obtained here demonstrate that Gadd45-induced cell cycle arrest in G2/M phase can significantly enhance TGE in CHO cells.
Publisher
KOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
Issue Date
2014-06
Language
English
Article Type
Article
Keywords

RECOMBINANT PROTEIN-PRODUCTION; MAMMALIAN-CELLS; CHO-CELLS; MONOCLONAL-ANTIBODIES; P53-REGULATED PROTEIN; KINASE-ACTIVITY; TRANSFECTION; APOPTOSIS; GADD45; PRODUCTIVITY

Citation

BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, v.19, no.3, pp.386 - 393

ISSN
1226-8372
DOI
10.1007/s12257-014-0151-0
URI
http://hdl.handle.net/10203/192708
Appears in Collection
BS-Journal Papers(저널논문)
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