Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems.
Cell expansion, gene transfer and protein production were all executed with a single serum-free, animal protein-free commercial medium designed for suspension-adapted Chinese hamster ovary cells (CHO DG44). This is a most important process to consider for clinical production of recombinant proteins. The transfection with polyethylenimine (PEI) was shown here to be scalable using both stirred-tank bioreactors of 3- and 150-l and novel agitated cultivation vessels (50 ml ventilated centrifuge tubes and 1-l square-shaped glass bottles) that lack any instrumentation. The transient transfections spanned a range of working volumes from 2 ml to 80 l. The maximum transient recombinant antibody yield was 22 mg/l, the highest ever reported for a multiliter transfection in CHO. The transiently expressed protein had the same extent of glycosylation as the same antibody produced from a stably transfected recombinant CHO cell line.
Muller
Natalie
N
Ecole Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland.
Derouazi
Madiha
M
Van Tilborgh
Frédéric
F
Wulhfard
Sarah
S
Hacker
David L
DL
Jordan
Martin
M
Wurm
Florian M
FM
D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006031:Glycosylation / Q000187:drug effects; D007074:Immunoglobulin G / Q000378:metabolism; D011094:Polyethyleneimine / Q000494:pharmacology; D014162:Transfection
D007074:Immunoglobulin G; D011094:Polyethyleneimine; D004247:DNA