title: Rapid recombinant protein production from piggyBac transposon-mediated stable CHO cell pools.
abstract: Heterogeneous populations of stably transfected cells (cell pools) can serve for the rapid production of moderate amounts of recombinant proteins. Here, we propose the use of the piggyBac (PB) transposon system to improve the productivity and long-term stability of cell pools derived from Chinese hamster ovary (CHO) cells. PB is a naturally occurring genetic element that has been engineered to facilitate the integration of a transgene into the genome of the host cell. In this report PB-derived cell pools were generated after 10 days of selection with puromycin. The resulting cell pools had volumetric productivities that were 3-4 times higher than those achieved with cell pools generated by conventional plasmid transfection even though the number of integrated transgene copies per cell was similar in the two populations. In 14-day batch cultures, protein levels up to 600 and 800 mg/L were obtained for an Fc-fusion protein and a monoclonal antibody, respectively, at volumetric scales up to 1L. In general, the volumetric protein yield from cell pools remained constant for up to 3 months in the absence of selection. In conclusion, transfection of CHO cells with the PB transposon system is a simple, efficient, and reproducible approach to the generation of cell pools for the rapid production of recombinant proteins.
authors.affiliation: Laboratory for Cellular Biotechnology, CH-1015 Lausanne, Switzerland.
authors.forename: Sowmya
authors.initials: S
authors.lastname: Balasubramanian
authors.affiliation: Laboratory for Cellular Biotechnology, CH-1015 Lausanne, Switzerland.
authors.forename: Mattia
authors.initials: M
authors.lastname: Matasci
authors.affiliation: Polymers Laboratory, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Kadlecova
authors.affiliation: Laboratory for Cellular Biotechnology, CH-1015 Lausanne, Switzerland.
authors.forename: Lucia
authors.initials: L
authors.lastname: Baldi
authors.affiliation: Laboratory for Cellular Biotechnology, CH-1015 Lausanne, Switzerland; Protein Expression Core Facility, CH-1015 Lausanne, Switzerland.
authors.forename: David L
authors.initials: DL
authors.lastname: Hacker
authors.affiliation: Laboratory for Cellular Biotechnology, CH-1015 Lausanne, Switzerland. Electronic address: florian.wurm@epfl.ch.
authors.forename: Florian M
authors.initials: FM
authors.lastname: Wurm
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004251:DNA Transposable Elements / Q000235:genetics*; D000068800:Etanercept / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; D011994:Recombinant Proteins; D000068800:Etanercept
keywords: CHO cells; Cell pool; Recombinant protein; Transfection; Transposon; piggyBac