Piggybac transposon-based production of anti-HER2 CAR T cell

Results: The HEK 293 FT cell line were analyzed 24 hours after the transfection with 2 ug of transposon, exhibiting an expression of 31,3% and 14,9% for 4D5 and FRP5 CARs respectively. For the JUKART cell line, we used 5ug, 10ug or 20ug PB plasmid concentrations and evaluated CAR expression 24 hours after transduction. We noted a higher expression of the receptors when using 10 ug of plasmid, obtaining 35,6% and 38,1% positivity for 4D5 and FRP5 receptors respectively. In the case of the primary T cells, were used an 10ug:5ug transposon:transposase concentration ratio gene-modifying cells from 3 different healthy donors, and the CAR expression was analyzed 1, 6 and 9 days post electroporation. The cells exhibited a decrease in the 4D5 and FRP5 receptors expression from day 1 to 6 after electroporation, getting more stable by day 9, with 8,94% and 5,06% average expression of 4D5 and FRP5 CARs respectively.