TY - JOUR
T1 - Simultaneous environmental manipulations in semi-perfusion cultures of CHO cells producing rh-tPA
AU - Vergara, Mauricio
AU - Becerra, Silvana
AU - Díaz-Barrera, Alvaro
AU - Berrios, Julio
AU - Altamirano, Claudia
PY - 2012/11/11
Y1 - 2012/11/11
N2 - We evaluated the combined effect of decreasing the temperature to a mild hypothermia range (34 and 31°C) 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 34°C and 55% at 31°C, compared to cultures at 37°C. Similar behaviour was observed in cultures at 34°C using galactose as a carbon source. Nonetheless, at 31°C, the specific productivity of rh-tPA strongly decreased (about 58%) compared to the culture at 37°C. In semi-perfusion culture, the highest rh-tPA specific productivity was obtained at 34°C. 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 37°C 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.
AB - We evaluated the combined effect of decreasing the temperature to a mild hypothermia range (34 and 31°C) 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 34°C and 55% at 31°C, compared to cultures at 37°C. Similar behaviour was observed in cultures at 34°C using galactose as a carbon source. Nonetheless, at 31°C, the specific productivity of rh-tPA strongly decreased (about 58%) compared to the culture at 37°C. In semi-perfusion culture, the highest rh-tPA specific productivity was obtained at 34°C. 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 37°C 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.
KW - CHO cells
KW - Galactose
KW - Glutamate
KW - Low temperature
KW - Perfusion culture
KW - Rh-tPA
UR - http://www.scopus.com/inward/record.url?scp=84870021614&partnerID=8YFLogxK
U2 - 10.2225/vol15-issue6-fulltext-2
DO - 10.2225/vol15-issue6-fulltext-2
M3 - Article
AN - SCOPUS:84870021614
VL - 15
JO - Electronic Journal of Biotechnology
JF - Electronic Journal of Biotechnology
SN - 0717-3458
IS - 6
ER -