Whilst none of the antifoams affected the growth rate of the cells, addition of P2000 or SB2121 was found to increase culture density. When normalized to the culture density, the GFP specific yield (μg OD 595 -1) was only increased for Antifoam A, Antifoam C and J673A. pastoris increased the total amount of recombinant GFP in the culture medium (the total yield) and in the case of P2000, SB2121 and J673A almost doubled it. ResultsĪddition of defined concentrations of Antifoam A (Sigma), Antifoam C (Sigma), J673A (Struktol), P2000 (Fluka) or SB2121 (Struktol) to shake-flask cultures of P. In this study we characterised, for the first time, the effects of five commonly-used antifoaming agents on the total amount of recombinant green fluorescent protein (GFP) secreted from shake-flask cultures of this industrially-relevant yeast. Intriguingly, antifoams are often added without prior consideration of their effect on the yeast cells, the protein product or the influence on downstream processes such as protein purification. A particular problem in these formats is foaming, which is commonly prevented by the addition of chemical antifoaming agents. This is followed by up-scaling either to shake-flasks or continuously stirred tank bioreactors. Initial screening for both suitable clones and optimum culture conditions is typically carried out in multi-well plates. Pichia pastoris is a widely-used host for recombinant protein production.
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