Biotechnology and Bioengineering, Vol.69, No.3, 266-274, 2000
Enhanced productivity during controlled proliferation of BHK cells in continuously perfused bioreactors
A perfused cell-culture process was developed to investigate the stability of IRF-1-mediated proliferation control in BHK cells and to evaluate the efficacy of a novel promoter in these cells. The cell density of proliferation-controlled producer cells was effectively regulated for over 7 weeks in a microcarrier-based continuously perfused bioreactor. An IRF-1-inducible promoter was employed to express a heterodimeric IgG antibody as a relevant model protein. Basal expression levels were equivalent to that of a highly active viral promoter, while productivity increased up to sixfold during growth arrest. However, no stably expressing clone was isolated in this study. Protein expression decreased gradually with time and could not be induced further in subsequent growth-repression cycles. The results demonstrate that the regulator/ system is sufficiently stable to allow controlled growth in a continous scalable reactor system and that productivity increases can be achieved in a proliferation controlled microcarrier culture.
Keywords:proliferation control;recombinant IgG antibody production;dicistronic mRNA;baby hamster kidney;(BHK) cells;IRF-1;microcarrier