Genomic integration as a way to stable high producers CHO cells

Abstract

Methodology: This methodology is based on DNA genomic integration by sleeping beauty transposase. Transfections were made with 25 μg of PEI (polyethylenimine) and 12,5 μg of total DNA (75% of DNA of interest and 25% of transposase DNA) for a total of 107 cells. Two days after, transfectants were selected by FACS, an antibiotic independent process, using GFP as the reporter gene (controled by IRES). Two more sortings rounds were necessary to stablish a 100% transfectant stable population. Cell culture supernatant was collected, and protein of interest was analysed by Western blot and ELISA.