About making a CHO production cell line “research-friendly” by genetic engineering

Voedisch, Bernd; Patoux, Agnes; Sterkenburgh, Jildou; Buchs, Mirjam; Barry, Emily; Allard, Cyril; Geisse, Sabine

doi:10.1186/1753-6561-5-s8-p132

Cited by 4 publications

(1 citation statement)

References 4 publications

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“…However, Jones et al demonstrated cell‐specific cytotoxicity associated with EBNA‐1 degradation. Voedisch et al, whilst creating a stable CHO‐EBNA‐1 cell line, found the majority of screened clones to contain degraded EBNA‐1—this suggests that EBNA‐1 might be a difficult protein to express stably in CHO cells. A possible mechanism of EBNA‐1 cytotoxicity is genomic instability that has been observed in tumor cells resulting from the induction of oxidative stress leading to DNA double‐strand breaks, engagement of the DNA damage response, and chromosomal aberrations .…”

Section: Resultsmentioning

confidence: 99%

A high‐yielding CHO transient system: Coexpression of genes encoding EBNA‐1 and GS enhances transient protein expression

Daramola¹,

Stevenson²,

Dean³

et al. 2013

Biotechnology Progress

127

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An efficient rapid protein expression system is crucial to support early drug development. Transient gene expression is an effective route, and to facilitate the use of the same host cells as for subsequent stable cell line development, we have created a high-yielding Chinese hamster ovary (CHO) transient expression system. Suspension-adapted CHO-K1 host cells were engineered to express the gene encoding Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) with and without the coexpression of the gene for glutamine synthetase (GS). Analysis of the transfectants indicated that coexpression of EBNA-1 and GS enhanced transient expression of a recombinant antibody from a plasmid carrying an OriP DNA element compared to EBNA-1-only transfectants. This was confirmed with the retransfection of an EBNA-1-only cell line with a GS gene. The retransfected cell lines showed an increase in transient expression when compared with that of the EBNA-1-only parent. The transient expression process for the best CHO transient cell line was further developed to enhance protein expression and improve scalability by optimizing the transfection conditions and the cell culture process. This resulted in a scalable CHO transient expression system that is capable of expressing 2 g/L of recombinant proteins such as antibodies. This system can now rapidly provide gram amounts of recombinant antibody to supply preclinical development studies that has comparable product quality to antibody produced from a stably transfected CHO cell line.

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Section: Resultsmentioning

confidence: 99%

A high‐yielding CHO transient system: Coexpression of genes encoding EBNA‐1 and GS enhances transient protein expression

Daramola¹,

Stevenson²,

Dean³

et al. 2013

Biotechnology Progress

127

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Engineering the supply chain for protein production/secretion in yeasts and mammalian cells

Klein

Niklas

Heinzle

2015

Journal of Industrial Microbiology and Biotechnology

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Metabolic bottlenecks play an increasing role in yeasts and mammalian cells applied for high-performance production of proteins, particularly of pharmaceutical ones that require complex posttranslational modifications. We review the present status and developments focusing on the rational metabolic engineering of such cells to optimize the supply chain for building blocks and energy. Methods comprise selection of beneficial genetic modifications, rational design of media and feeding strategies. Design of better producer cells based on whole genome-wide metabolic network analysis becomes increasingly possible. High-resolution methods of metabolic flux analysis for the complex networks in these compartmented cells are increasingly available. We discuss phenomena that are common to both types of organisms but also those that are different with respect to the supply chain for the production and secretion of pharmaceutical proteins.

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Tissue decellularization by activation of programmed cell death

et al. 2013

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About making a CHO production cell line “research-friendly” by genetic engineering

Cited by 4 publications

References 4 publications

A high‐yielding CHO transient system: Coexpression of genes encoding EBNA‐1 and GS enhances transient protein expression

A high‐yielding CHO transient system: Coexpression of genes encoding EBNA‐1 and GS enhances transient protein expression

Engineering the supply chain for protein production/secretion in yeasts and mammalian cells

Tissue decellularization by activation of programmed cell death

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