Marchant, R.J. and Al-Fageeh, M.B. and Underhill, M.F. and Racher, A.J. and Smales, C.M. (2008) Metabolic rates, growth phase, and mRNA levels influence cell-specific antibody production levels from in vitro-cultured mammalian cells at sub-physiological temperatures. Molecular Biotechnology, 39 (1). pp. 69-77. ISSN 1073-6085.
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Previous work has shown that recombinant protein yield can be improved from in vitro-cultured mammalian cells by culturing at sub-physiological temperatures, although this effect is cell line and product dependent. The mechanism(s) by which low temperature leads to enhanced product yield are currently unknown; however, recent reports suggest that increased mRNA levels at sub-physiological temperatures may be largely responsible for this. Here, we have investigated whether low-temperature cultivation of cell lines selected for antibody production at 37C leads to changes in heavy- and light-chain mRNA levels and if this is reflected in antibody yields. Low-temperature in vitro culturing resulted in reduced viable cell concentration, prolonged cell viability, a reduction in metabolite consumption and production, cell cycle arrest in both CHO and NS0 cells, and changes in the levels of heavy- and light-chain mRNA. Despite increases in the level of heavy- and light-chain mRNA upon culturing at 32C in our model CHO cell line, this did not result in increased total product yield; however, changes in cell-specific yields were observed that reflected the metabolic rate of glucose utilization and changes in mRNA levels.
|Uncontrolled keywords:||cold-shock; sub-physiological temperature culturing; CHO; NS0; mRNA levels; monoclonal antibody production|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Mark Smales|
|Date Deposited:||14 Apr 2009 13:53|
|Last Modified:||14 Jan 2010 14:23|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/6234 (The current URI for this page, for reference purposes)|
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