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  4. Rapid recombinant protein production from piggyBac transposon-mediated stable CHO cell pools
 
research article

Rapid recombinant protein production from piggyBac transposon-mediated stable CHO cell pools

Balasubramanian, Sowmya
•
Matasci, Mattia
•
Kadlecova, Zuzana
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2015
Journal of Biotechnology

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 1 L. 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.

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Type
research article
DOI
10.1016/j.jbiotec.2015.03.001
Web of Science ID

WOS:000352017700012

PubMed ID

25758242

Author(s)
Balasubramanian, Sowmya
Matasci, Mattia
Kadlecova, Zuzana
Baldi, Lucia  
Hacker, David L.  
Wurm, Florian M.  
Date Issued

2015

Publisher

Elsevier

Published in
Journal of Biotechnology
Volume

200

Start page

61

End page

69

Subjects

Recombinant protein

•

piggyBac

•

Transposon

•

Cell pool

•

Transfection

•

CHO cells

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBTC  
Available on Infoscience
May 19, 2015
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/113975
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