title: Foreign protein expression from S phase specific promoters in continuous cultures of recombinant CHO cells.
abstract: Foreign protein expression from the commonly used SV40 promoter has been found to be primarily during the S-phase of the cell cycle. Simple mathematical models with this cell cycle phase dependent expression of foreign protein suggest that the specific production rate will be proportional to the cell growth rate, which is particularly disadvantageous in high cell density fed-batch or perfusion bioreactors. In this study we investigate this predicted relationship between the production rate and growth rate by culturing recombinant CHO cells in a continuous suspension bioreactor. One CHO cell line, GS-26, has been stably transfected with the plasmid pSVgal, which contains the E. coli lac Z gene under the control of the SV40 promoter. This GS-26 cell line was grown in suspension cultures over a range of specific growth rates in batch and continuous modes. The intracellular β-galactosidase activity was assayed using a standard spectrophotometric method after breaking the cells open and releasing the enzyme. A strong growth associated relationship is found between the intracellular β-galactosidase content and the specific growth rate in batch and continuous cultures, as predicted.
authors.affiliation: Department of Chemical Engineering, University of Colorado, 80309-0424, Boulder, CO, USA.
authors.forename: G G
authors.initials: GG
authors.lastname: Banik
authors.forename: P W
authors.initials: PW
authors.lastname: Todd
authors.forename: D S
authors.initials: DS
authors.lastname: Kompala