Overexpression of bcl-2 inhibits sodium butyrate-induced apoptosis in Chinese hamster ovary cells resulting in enhanced humanized antibody production

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Sodium butyrate (NaBu) can enhance the expression of genes from some of the mammalian promoters including cytomegalovirus (CMV) and simian virus 40 (SV40), but it can also inhibit cell growth and induce cellular apoptosis. Thus, the beneficial effect of using a higher concentration of NaBu on a foreign protein expression is compromised by its cytotoxic effect on cell growth. To overcome this cytotoxic effect of NaBu, a survival protein, human Bcl-2, was overexpressed in recombinant Chinese hamster ovary (CHO) cells (SH2-0.32), producing a humanized antibody directed against the S surface antigen of hepatitis B virus. When batch cultures of both control cells transfected with bcl-2-deficient plasmid (SH2-0.32-Δbcl-2) and cells transfected with bcl-2 expression plasmid (14C6-bcl-2) were performed in the absence of NaBu, both cells showed similar profiles of cell viability and antibody production. Compared with the SH2-0.32-Δbcl-2 culture, under the condition of NaBu addition at the exponential growth phase, overexpression of the bcl-2 gene considerably suppressed the NaBu-induced apoptosis of 14C6-bcl-2 by inhibiting caspase 3 activity and extending culture longevity by >2 days. As a result, the final antibody concentration of 14C6-bcl-2 culture was twofold higher than that of SH2-0.32-Δbcl-2 culture in the presence of NaBu and threefold higher than that of SH2-0.32-Δbcl-2 and 14C6-bcl-2 cultures in the absence of NaBu. © 2001 John Wiley & Sons, Inc.
Publisher
Wiley-VCH Verlag
Issue Date
2001-08
Language
English
Article Type
Article
Keywords

HIGH-LEVEL EXPRESSION; CHIMERIC ANTIBODY; GENE-EXPRESSION; CULTURE CONDITIONS; HYBRIDOMA CELLS; BATCH CULTURES; CHO CELLS; DEATH; GROWTH; CYCLE

Citation

BIOTECHNOLOGY AND BIOENGINEERING, v.71, no.3, pp.184 - 193

ISSN
0006-3592
DOI
10.1002/1097-0290(2000)71:3<184::AID-BIT1008>3.0.CO;2-W
URI
http://hdl.handle.net/10203/77648
Appears in Collection
BS-Journal Papers(저널논문)
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