Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions

Mercille, Sylvain; Massie, Bernard

doi:10.1002/(sici)1097-0290(19990605)63:5<529::aid-bit3>3.0.co;2-x

Cited by 49 publications

(26 citation statements)

References 99 publications

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“…The most common genetic modification reported in literature involves over-expression of either Bcl-2 or Bcl-xl, although other genes such as E1B19K, XIAP and Bhrf-1 (Mercille and Massie 1999;Sauerwald et al 2003;Juanola et al 2009) has also been used to confer apoptosis resistance. The bcl-2 gene (2009) Bcl-xl Bcl-2-like 1 CHO was the first to be recognized as a survival gene involved in the apoptotic pathway and demonstrated increase in cell density and viability correlated with delay of apoptosis in hybridoma (Itoh et al 1995;Al-Rubeai et al 1995a).…”

Section: Apoptosis Engineeringmentioning

confidence: 99%

“…Apparently in all the above studies bcl-2 homologs demonstrated varying degree of protection depending on the nature and strength of insult which may be partially explained by the anti-apoptotic gene expression level at the time of insult which was shown to vary with time during batch culture (results not published), cell to cell or clonal variability resulted from genetic instability or functionally redundancy of the gene presumably due to their low expression. Indeed, a threshold level of expression was reported to be required E1B-19K (Mercille and Massie 1999) activity to protect cells from apoptosis.…”

Section: Apoptosis Engineeringmentioning

confidence: 99%

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Cell death in mammalian cell culture: molecular mechanisms and cell line engineering strategies

2010

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Cell death is a fundamentally important problem in cell lines used by the biopharmaceutical industry. Environmental stress, which can result from nutrient depletion, by-product accumulation and chemical agents, activates through signalling cascades regulators that promote death. The best known key regulators of death process are the Bcl-2 family proteins which constitute a critical intracellular checkpoint of apoptosis cell death within a common death pathway. Engineering of several members of the anti-apoptosis Bcl-2 family genes in several cell types has extended the knowledge of their molecular function and interaction with other proteins, and their regulation of cell death. In this review, we describe the various modes of cell death and their death pathways at molecular and organelle level and discuss the relevance of the growing knowledge of anti-apoptotic engineering strategies to inhibit cell death and increase productivity in mammalian cell culture.

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Section: Apoptosis Engineeringmentioning

confidence: 99%

Section: Apoptosis Engineeringmentioning

confidence: 99%

Cell death in mammalian cell culture: molecular mechanisms and cell line engineering strategies

2010

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“…During apoptosis, a cell population with a reduced DNA content develops and this population is accepted as an indicator of apoptotic cell death (Mercille and Massie, 1999). Al-Rubeai et al (1995) demonstrated that this cell population with a reduced DNA content, a sub-G 1 population, is indeed composed of apoptotic bodies.…”

Section: Overexpression Of Bcl-2 Gene Inhibits the Entrance Of Nabu-ementioning

confidence: 99%

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

Kim

Lee

2000

Biotechnol. Bioeng.

117

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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-⌬bcl-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-⌬bcl-2 culture, under the condition of NaBu addition at the exponential growth phase, overexpression of the bcl-2 gene considerably suppressed the NaBuinduced 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-⌬bcl-2 culture in the presence of NaBu and threefold higher than that of SH2-0.32-⌬bcl-2 and 14C6-bcl-2 cultures in the absence of NaBu.

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“…Some of the first studies showed that when hybridoma cells over-expressed the Bcl-2 protein it significantly increases cell viability and productivity (Simpson et al 1997). When myeloma cells, in perfusion culture, were engineered to express E1B-19K (the adenovirus Bcl-2 homologue) the resulted 2-fold increase in viable cell densities allowed for a 40% increase in monoclonal antibody yield (Mercille and Massie 1999). Another approach utilized small interfering RNA (siRNA) (Brummelkamp et al 2002;Elbashir et al 2001) constructs directed at mRNA sequences of two pro-apoptotic proteins, Bax and Bak (Lim et al 2006).…”

Section: Apoptosis Engineeringmentioning

confidence: 99%

Using cell engineering and omic tools for the improvement of cell culture processes

et al. 2007

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Significant strides have been made in mammalian cell based biopharmaceutical process and cell line development over the past years. With several established mammalian host cell lines and expression systems, optimization of selection systems to reduce development times and improvement of glycosylation patterns are only some of the advances being made to improve cell culture processes. In this article, the advances pertaining to cell line development and cell engineering strategies are discussed. An overview of the cell engineering strategies to enhance cellular characteristics by genetic manipulation are illustrated, focusing on the use of genomics and proteomics tools and their application in such endeavors. Included in this review are some of the early studies using the 'omic' technique to understand cellular mechanisms of product synthesis and secretion, apoptosis, cell proliferation and the influence of the physicochemical environment. The article highlights the significance of integrating genomics and proteomics data with the vast amounts of bioprocess data for improved analysis of the biological pathways involved. Further improvements of the techniques and methodologies used are needed but ultimately, the new cell engineering strategies should provide great insight into the regulatory networks within the cell in a bioprocess environment and how to manipulate them to increase overall productivity.

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Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions

Cited by 49 publications

References 99 publications

Cell death in mammalian cell culture: molecular mechanisms and cell line engineering strategies

Cell death in mammalian cell culture: molecular mechanisms and cell line engineering strategies

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

Using cell engineering and omic tools for the improvement of cell culture processes

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