p27Kip1-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 B11 cells adapted for growth in serum-free suspension cultures for unlinked muristerone-inducible expression of the cyclin-dependent kinase inhibitor p27Kip1 and constitutive expression of the soluble intercellular adhesion molecule-1 (sICAM), a potent common cold therapeutic. Conditional overexpression of p27Kip1 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.
Meents
Heiko
H
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich.
Enenkel
Barbara
B
Werner
Rolf G
RG
Fussenegger
Martin
M
D000818:Animals; D016466:CHO Cells / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D018797:Cell Cycle Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D002455:Cell Division / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004305:Dose-Response Relationship, Drug; D004441:Ecdysterone / Q000008:administration & dosage* / Q000031:analogs & derivatives*; D004791:Enzyme Inhibitors; D005786:Gene Expression Regulation / Q000187:drug effects*; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013535:Suspensions; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics
D018797:Cell Cycle Proteins; D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D013535:Suspensions; D025521:Tumor Suppressor Proteins; D018799:Intercellular Adhesion Molecule-1; D050760:Cyclin-Dependent Kinase Inhibitor p27; C055348:muristerone A; D004441:Ecdysterone; D013762:Tetrahydrofolate Dehydrogenase