Metabolic rates, growth phase, and mRNA levels influence cell-specific antibody production levels from in vitro-cultured mammalian cells at sub-physiological temperatures

Marchant, Rosalyn J. and Al-Fageeh, Mohamed B. and Underhill, Michele F. and Racher, Andrew J. and Smales, C. Mark (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. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1007/s12033-008-9032-0

Abstract

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.

Item Type: Article
Uncontrolled keywords: cold-shock; sub-physiological temperature culturing; CHO; NS0; mRNA levels; monoclonal antibody production
Subjects: Q Science
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: Mark Smales
Date Deposited: 14 Apr 2009 13:53
Last Modified: 09 Apr 2014 09:00
Resource URI: http://kar.kent.ac.uk/id/eprint/6234 (The current URI for this page, for reference purposes)
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