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Differential response in downstream processing of CHO cells grown under mild hypothermic conditions.

Tait, AS; Tarrant, RD; Velez-Suberbie, ML; Spencer, DI; Bracewell, DG; (2013) Differential response in downstream processing of CHO cells grown under mild hypothermic conditions. Biotechnology Progress , 29 (3) pp. 688-696. 10.1002/btpr.1726. Green open access

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Abstract

The manufacture of complex therapeutic proteins using mammalian cells is well established, with several strategies developed to improve productivity. The application of sustained mild hypothermic conditions during culture has been associated with increases in product titer and improved product quality. However, despite associated cell physiological effects, very few studies have investigated the impact on downstream processing (DSP). Characterization of cells grown under mild hypothermic conditions demonstrated that the stationary phase was prolonged by delaying the onset of apoptosis. This enabled cells to maintain viability for extended periods and increase volumetric productivity from 0.74 to 1.02 g L(-1) . However, host cell proteins, measured by ELISA, increased by ∼50%, attributed to the extended time course and higher peak and harvest cell densities. The individual components making up this impurity, as determined by SELDI-TOF MS and 2D-PAGE, were shown to be largely comparable. Under mild hypothermic conditions, cells were less shear sensitive than those maintained at 37°C, enhancing the preliminary primary recovery step. Adaptive changes in membrane fluidity were further investigated by adopting a pronounced temperature shift immediately prior to primary recovery and the improvement observed suggests that such a strategy may be implementable when shear sensitivity is of concern. Early and late apoptotic cells were particularly susceptible to shear, at either temperature, even under the lowest shear rate investigated. These findings demonstrate the importance of considering the impact of cell culture strategies and cell physiology on DSP, by implementing a range of experimental methods for process characterization. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 2013.

Type: Article
Title: Differential response in downstream processing of CHO cells grown under mild hypothermic conditions.
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.1726
Publisher version: http://dx.doi.org/10.1002/btpr.1726
Language: English
Additional information: This work is licensed under the Creative Commons Attribution License, Attribution 3.0 Unported (CC BY 3.0) http://creativecommons.org/licenses/by/3.0/ which permits unrestricted use, distribution, and reproduction in any medium. However, you must attribute the work to the author (but not in any way that suggests that they endorse you or your use of the work).
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1392567
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