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Biotechnol Bioeng. 2016 Jun;113(6):1234-43. doi: 10.1002/bit.25888. Epub 2015 Dec 9.

Comparison of three transposons for the generation of highly productive recombinant CHO cell pools and cell lines.

Author information

1
Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
2
Protein Expression Core Facility, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
3
Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland. florian.wurm@epfl.ch.

Abstract

Several naturally occurring vertebrate transposable elements have been genetically modified to enable the transposition of recombinant genes in mammalian cells. We compared three transposons-piggyBac, Tol2, and Sleeping Beauty-for their ability to generate cell pools (polyclonal cultures of recombinant cells) and clonal cell lines for the large-scale production of recombinant proteins using Chinese hamster ovary cells (CHO-DG44) as the host. Transfection with each of the dual-vector transposon systems resulted in cell pools with volumetric yields of tumor necrosis factor receptor-Fc fusion protein (TNFR:Fc) that were about ninefold higher than those from cell pools generated by conventional plasmid transfection. On average, the cell pools had 10-12 integrated copies of the transgene per cell. In the absence of selection, the volumetric productivity of the cell pools decreased by 50% over a 2-month cultivation period and then remained constant. The average volumetric TNFR:Fc productivity of clonal cell lines recovered from cell pools was about 25 times higher than that of cell lines generated by conventional transfection. In 14-day fed-batch cultures, TNFR:Fc levels up to 900 mg/L were obtained from polyclonal cell pools and up to 1.5 g/L from clonal cell lines using any of the three transposons. Biotechnol. Bioeng. 2016;113: 1234-1243.

KEYWORDS:

Sleeping Beauty; Tol2; cell pools; piggyBac; recombinant protein; transposon

PMID:
26616356
DOI:
10.1002/bit.25888
[Indexed for MEDLINE]

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