Biotechnology and Bioengineering, Vol.79, No.6, 619-627, 2002
p27(Kip1)-mediated controlled proliferation technology increases constitutive sICAM production in CHO-DUKX adapted for growth in suspension and serum-free media
We have engineered dihydrofolate reductase-deficient (dhfr(-)) Chinese hamster ovary (CHO)-DUKX 1311 cells adapted for growth in serum-free suspension cultures for unlinked muristerone-inducible expression of the cyclin-dependent kinase inhibitor p27(Kip1) and constitutive expression of the soluble intercellular adhesion molecule-1 (sICAM), a potent common cold therapeutic. Conditional overexpression of p27(Kip1) resulted in a sustained G1-specific growth arrest of transgenic CHO-DUKX associated with up to fivefold-increased specific sICAM productivity. Herein we exemplify the implementation of controlled proliferation technology in a major biopharmaceutical production cell line that is compatible with key requirements for large-scale production procedures, including constitutive transgene expression and anchorage-independent growth in serum-free media.
Keywords:controlled proliferation;cyclin-dependent kinase inhibitor;p27;cell cycle;sICAM;serum-free;gene switch