000207935 001__ 207935
000207935 005__ 20190317000154.0
000207935 0247_ $$2doi$$a10.1016/j.jbiotec.2015.03.001
000207935 022__ $$a0168-1656
000207935 02470 $$2PMID$$a25758242
000207935 02470 $$2ISI$$a000352017700012
000207935 037__ $$aARTICLE
000207935 245__ $$aRapid recombinant protein production from piggyBac transposon-mediated stable CHO cell pools
000207935 260__ $$bElsevier$$c2015$$aAmsterdam
000207935 269__ $$a2015
000207935 300__ $$a9
000207935 336__ $$aJournal Articles
000207935 520__ $$aHeterogeneous 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.
000207935 6531_ $$aRecombinant protein
000207935 6531_ $$apiggyBac
000207935 6531_ $$aTransposon
000207935 6531_ $$aCell pool
000207935 6531_ $$aTransfection
000207935 6531_ $$aCHO cells
000207935 700__ $$aBalasubramanian, Sowmya
000207935 700__ $$aMatasci, Mattia
000207935 700__ $$aKadlecova, Zuzana
000207935 700__ $$0240397$$g117359$$aBaldi, Lucia
000207935 700__ $$0240882$$g152132$$aHacker, David L.
000207935 700__ $$aWurm, Florian M.$$g107554$$0240398
000207935 773__ $$j200$$tJournal of Biotechnology$$q61-69
000207935 8564_ $$uhttp://www.sciencedirect.com/science/article/pii/S0168165615000954$$zURL
000207935 909C0 $$xU10180$$0252009$$pLBTC
000207935 909CO $$qGLOBAL_SET$$particle$$ooai:infoscience.tind.io:207935
000207935 917Z8 $$x117359
000207935 937__ $$aEPFL-ARTICLE-207935
000207935 973__ $$rREVIEWED$$sPUBLISHED$$aEPFL
000207935 980__ $$aARTICLE