Nutrient enrichment and in-situ waste removal through electrical means for hybridoma cultures.
In-situ dc electric fields were applied to remove ammonium and lactate from suspension hybridoma cultures (ATCC-CRL-1606) which used enriched media. Nutrient concentration was increased fourfold above the normal concentration of DMEM to study enhanced protein product formation in a dc electric field. In the presence of the electric field, hybridoma growth and antibody production were increased 1.5-fold (from 3.7 x 10(6) to 9.1 x 10(6) viable cells/mL) and twofold (from 170 to 505 mg IgG/L), respectively, compared with the control. The effective removal of ammonium and lactate and increased concentrations of the various nutrients accounted for this enhancement. The enriched media caused the overflow metabolism of glucose, glutamine, and various essential amino acids. The overconsumption of glucose also produced substantial amounts of lactate, which in turn greatly increased the medium osmolarity. The increase in medium osmolarity is believed to be one of the causes of cell death in these culture systems.(c) 1995 John Wiley & Sons, Inc.
Chang
Y H
YH
Department of Chemical Engineering, Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
Grodzinsky
A J
AJ
Wang
D I
DI