Effect of elevated oxygen and glutamine levels on foreign protein production at high cell densities using the insect cell-baculovirus expression system

Autor: Ronald A. Taticek, Michael L. Shuler
Rok vydání: 1997
Předmět:
Zdroj: Biotechnology and Bioengineering. 54:142-152
ISSN: 1097-0290
0006-3592
DOI: 10.1002/(sici)1097-0290(19970420)54:2<142::aid-bit6>3.0.co;2-l
Popis: Per cell protein expression in virally-infected insect cells declines significantly at high cell density resulting in a decrease in volumetric productivity. Specific protein expression levels in Spodoptera frugiperda (Sf-21) cells could be increased at high cell densities by increasing the oxygen supply and by supplementing the medium with glutamine post-infection. β-Galactosidase yield was increased from 411 to 855 IU/ml by increasing the glutamine concentration in the medium by 46% and increasing the gas phase oxygen concentration from 21 to 80%. Similarly, the yield of a secreted alkaline phosphatase was increased from 14.2 to 26.2 IU/mL using the same conditions. Part of the increase in production with Sf-21 culture was due to increased release to the extra-cellular compartment at the higher oxygen concentrations. Increasing the gas phase oxygen concentration to 95% in conjunction with a 100% increase in glutamine and glucose concentrations did not improve the yield any further. Peak production under elevated oxygen and nutrient conditions occurred at 72 h about 24–48 h earlier than under normal conditions. In a Trichoplusia ni cell line (BTI-TN-5B1-4), the maximum secreted alkaline phosphatase activity was increased from 10 to 27.2 IU/mL by similarly manipulating the oxygen supply. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 142–152, 1997.
Databáze: OpenAIRE
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