title: Comparison of three transposons for the generation of highly productive recombinant CHO cell pools and cell lines.
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. © 2015 Wiley Periodicals, Inc.
authors.affiliation: Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Sowmya
authors.initials: S
authors.lastname: Balasubramanian
authors.affiliation: Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Yashas
authors.initials: Y
authors.lastname: Rajendra
authors.affiliation: Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Lucia
authors.initials: L
authors.lastname: Baldi
authors.affiliation: Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.|Protein Expression Core Facility, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: David L
authors.initials: DL
authors.lastname: Hacker
authors.affiliation: Laboratory of Cellular Biotechnology, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland. florian.wurm@epfl.ch.
authors.forename: Florian M
authors.initials: FM
authors.lastname: Wurm
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods*; D016466:CHO Cells; D003412:Cricetulus / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods*; D009419:Nerve Tissue Proteins / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics*; D019895:Transposases / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D009419:Nerve Tissue Proteins; C569369:PGBD1 protein, human; D011994:Recombinant Proteins; C062093:tolA protein, E coli; D019895:Transposases; C494471:sleeping beauty transposase, human
keywords: Sleeping Beauty; Tol2; cell pools; piggyBac; recombinant protein; transposon