Mead, E.J. and Chiverton, L.M. and Smales, C.M. and von der Haar, T. (2009) Identification of the limitations on recombinant gene expression in CHO cell lines with varying luciferase production rates. Biotechnology and Bioengineering, 102 (6). pp. 1593-1602. ISSN 0006-3592 .
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Mammalian cell lines are currently employed as one of the main cellular factories for the expression of recombinant protein-based drugs. The establishment of high-producing cell lines typically begins with a heterogeneous starter population of cells, from which the highest producing cells are selected via empirical approaches. This approach is time consuming, and is likely to encounter natural upper limits imposed by the inherent biology of the cell lines in question. In an attempt to understand both the nature of the variability in populations of cells transfected with recombinant protein encoding DNA and the natural mechanisms of productivity limitation, we developed protocols for the detailed investigation of gene expression pathways in such cell lines. This novel approach was then applied to a set of clonal CHOK1 cell lines producing recombinant luciferase with varying productivities. Our results show that the initial limitation in these cell lines is at the transcriptional level, however in the highest producing cell line post-translational mechanisms affecting both protein turnover and protein folding become severely limiting. The implications for the development of strategies to engineer cells for enhanced recombinant protein production levels are discussed.
|Uncontrolled keywords:||posttranscriptional control; translational control; protein turnover; recombinant protein; mammalian cell culture|
|Subjects:||Q Science > QP Physiology (Living systems) > QP506 Molecular biology|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences > Protein Science Group|
|Depositing User:||Tobias von der Haar|
|Date Deposited:||15 Sep 2009 09:20|
|Last Modified:||15 Sep 2009 09:20|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/22646 (The current URI for this page, for reference purposes)|
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