High Protein Yield - NEW: FectoPRO™

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FectoPROTM gives amazing antibody production yields in suspension CHO cells.
Cells were seeded at 1 x 106 cells/ml in 30ml of FreestyleTM CHO Expression Medium and transfected with FectoPROTM (0.8μg DNA/ml) or FreeStyleTM MAX Reagent (1.25μg DNA/,ml)
following recommended protocols for the respective reagents. Recombinant mouse lgG production yield was assayed 1 to 6 days after transfection using protein G biosensor (fortéBIO® octet RED96 system).

Data kindly provided by ProteoGenix SAS (France)

FectoPROTM gives high transfection efficiency in suspension CHO cells.
FreeStyleTM CHO-S cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyleTM CHO Expression Medium and transfected with FectoPROTM + FectoPROTM Booster of FreeStyleTM MAX Reagent using 0.4μg/ml and 1.25μg/,l of a GFP expressing plasmid, respectively. GFP expression was assayed 24 h after transfection using fluorescence microscopy.

FectoPROTM gives superior protein prodcuction yields in suspension CHO and HEK-293 cells.
FreeStyleTM CHO-S and HEK-293F cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyleTM CHO or 293 Expression Medium and transfected with FectioPROTM + FectoPROTM Booster (0.4μg DNA/ml), FreeStyle TM MAx Reagent (1.25μg DNA/ml) or PEIproTM (1μg DNA/ml) following recommended protocols for the respective reagents. IgG3-Fc (34kDa) production was assayed 72 h (CHO cells) and 120 h (HEK-293 cells) after transfection by protein G affinity quantification (HPLC) and Western Blot. Standard corresponds to purified IgG3-Fc (34kDa).

FectoPROTM HEK-293F cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyleTM 293 Expression Medium and transfected with FectoPROTM + FectoPROTM Booster (0.5 μg DNA/,l) following the standards protocol. IgG3-Fc production was assayed at different day by protein G affinity quantification (HPLC). Similar results are obtained with FreeStyleTM CHO-S (data not shown).

FectoPROTM is easily scalable and reproducible in CHO cells.
FreeStyleTM CHO-S cells were seeded at 1 x 106 cells/ml in 30ml, 100ml, 300ml or 1L of FreeStyleTM CHO Expression Medium and transfected with FectoPROTM + FectoPROTM Booster (0.5μg DNA/ml). At day 1 to 6, cell viability was assayed using Trypan blue exclusion assay (A) and IgG3-Fc production was assayed by protein G affinity quantification (HPLC) (B) and Western Blot (C). Standard (C) corresponds to purified IgG3-Fc (34 kDa).

Suspension CHO cells were seeded at 1 x 106 cells/ml in 30 ml of CHO medium and transfected using 3 different lots of FectoPROTM (CCIII 078, CCIII 104 or CCIII 106) following the standard protocol. 72 h after transfection, IgG3-Fc production was assayed by protein G affinity quantification. Similar results are obtained on suspension HEK-2933 cells (data not shown).

FectoPROTM gives superior protein production yields in suspension CHO cells grown in different media. CHO cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyleTM CHO Expression Medium, CD CHO Medium or Pro-CHOTM 4 Medium and transfected with FectoPROTM + FectPROTM Booster or FreeStyleTM MAX Reagent following recommended protocols for the respective reagents. IgG3-Fc production was assayed 72 h after transfection by protein G affinity quantification (HPLC).