Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems

Authors:
Muller N1, Derouazi M, Van Tilborgh F, Wulhfard S, Hacker DL, Jordan M, Wurm FM.
In:
Source: Biotechnol Lett
Publication Date: (2007)
Issue: 29 (5): 703-11
Research Area:
Gene Expression
Cells used in publication:
CHO-DG44
Species: hamster
Tissue Origin:
CHO-DG44 (DHFR-)
Species: hamster
Tissue Origin: ovarian
Culture Media:
Abstract
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.