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p27 Kip1 ‐mediated controlled proliferation technology increases constitutive sICAM production in CHO‐DUKX adapted for growth in suspension and serum‐free media
Author(s) -
Meents Heiko,
Enenkel Barbara,
Werner Rolf G.,
Fussenegger Martin
Publication year - 2002
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10322
Subject(s) - chinese hamster ovary cell , cell growth , transgene , cell culture , microbiology and biotechnology , intracellular , dihydrofolate reductase , biology , biochemistry , gene , genetics
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 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. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 619–627, 2002.