Accelerating stable recombinant cell line development by targeted integration
نویسندگان
چکیده
Introduction Targeted integration (TI) allows fast and reproducible genetic modification of well characterized previously tagged host cells thus generating producer cells with predictable qualities. Concurrently, timelines are cut by 50% compared to random integration (RI) based cell line development. In contrast to commonly low productivities of cell lines generated by TI, we developed a system for CHO cells leading to productivities of more than 1 g/L within weeks using the TurboCellTM platform. The system is based on CHO K1 cells that have been tagged with a GFP expression cassette flanked by recombinase recognition sites. Following GFP based FACS enrichment and cloning of the tagged cells, over 4000 clones were screened for growth, productivity, GFP expression stability and integration status of the GFP expression cassette. The best clones were selected to be used as “Master TurboCell” (MTC) host cell lines for recombinant cell line development.
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