Simultaneous environmental manipulations in semi-perfusion cultures of CHO cells producing rh-tPA
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Keywords

perfusion culture
galactose
low temperature
glutamate
CHO cells
rh-tPA

How to Cite

1.
Vergara M, Becerra S, Diaz-Barrera A, Berrios J, Altamirano C. Simultaneous environmental manipulations in semi-perfusion cultures of CHO cells producing rh-tPA. Electron. J. Biotechnol. [Internet]. 2012 Oct. 11 [cited 2024 Dec. 3];15(6). Available from: https://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/v15n6-2

Abstract

We evaluated the combined effect of decreasing the temperature to a mild hypothermia range (34 and 31ºC) and switching to a slowly metabolizable carbon source (glucose substituted by galactose) on the growth and production of a recombinant human tissue plasminogen activator (rh-tPA) by Chinese hamster ovary cells in batch and semi-perfusion cultures. In batch cultures using glucose as a carbon source, decreasing the temperature caused a reduction in cell growth and an increase in specific productivity of rh-tPA of 32% at 34ºC and 55% at 31ºC, compared to cultures at 37ºC. Similar behaviour was observed in cultures at 34ºC using galactose as a carbon source. Nonetheless, at 31ºC, the specific productivity of rh-tPA strongly decreased (about 58%) compared to the culture at 37ºC. In semi-perfusion culture, the highest rh-tPA specific productivity was obtained at 34ºC. Similarly, whether a decrease in the temperature is accompanied of the replacement of glucose by galactose, the rh-tPA specific productivity improved about 112% over that obtained in semi-perfusion culture carried out at 37ºC with glucose as the carbon source. A semi-perfusion culture strategy was implemented based on the combined effect of the chosen carbon source and low temperatures, which was a useful approach for enhance the specific productivity of the recombinant protein.

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