Regulation of cell cycle and productivity in NS0 cells by the over-expression of p21CIP1.
We have constructed NS0 myeloma cell lines that inducibly express the p21CIP1 cyclin dependent kinase inhibitor, using the Lacswitch system. Ectopic p21(CIP1) protein expression was rapidly induced within 12 h of addition of IPTG, causing G1-phase arrest and almost complete inhibition of cell proliferation. The production of a chimeric IgG4 antibody, expressed constitutively from an independent promoter, was found to be significantly increased by more than 4-fold in p21CIP1-arrested cells. This study demonstrates for the first time the successful construction of anchorage-independent and proliferation-controlled NS0 cell lines with enhanced secreted chimeric antibody production independent of the inducible promoter activity used to achieve cytostasis.
Watanabe
Shikiko
S
School of Chemical Engineering, University of Birmingham, Birmingham B15 2TT, United Kingdom.
Shuttleworth
John
J
Al-Rubeai
Mohamed
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D015153:Blotting, Western / Q000379:methods; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D002455:Cell Division / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D050759:Cyclin-Dependent Kinase Inhibitor p21; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors; D016213:Cyclins / Q000096:biosynthesis* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; C493956:CDKN1A protein, human; C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D007074:Immunoglobulin G; D018844:Cyclin-Dependent Kinases