TransIT-PRO® Transfection Reagent & Kit

High Efficiency Transfection of CHO-S Cells Using the TransIT-PRO® Transfection Reagent. Plasmid DNA encoding EGFP was delivered by transient transfection with the TransIT-PRO® Transfection Reagent (1:1) (reagent:DNA ratio, volume:weight) using 1 µg plasmid DNA per milliliter of culture and cell a density of 2 x 10⁶ cells/ml in the CHOgro® Expression Medium at the time of transfection.  FreeStyle™ CHO-S cells were cultured in CHOgro® Expression Medium  and plated into non-treated 6-well plates (2 ml/well) for transfection. (A) GFP levels and cell viability (propidium iodide) were measured 48 hours post-transfection using a Guava® easyCyte™ 5HT flow cytometer (EMD Millipore). (B) Images were captured using a Zeiss Axiovert inverted fluorescence microscope.

TransIT-PRO® Yields Multi-fold Increases in Antibody Titer in Conjunction with CHOgro® Expression Medium. Human IgG1 was produced by transient transfection using TransIT-PRO® (1:1), FreeStyle™ MAX (1.25:1.25) or 25kDa linear PEI (6:1) transfection reagents according to the manufacturers’ or published protocol (reagent:DNA ratio). Transfections were performed using 1 µg plasmid DNA per milliliter of culture and cell densities of 2 x 10⁶ cells/ml or 1x 10⁶ cells/ml for the CHOgro® Expression Medium (red bars) or FreeStyle™ Expression Medium (blue bars), respectively, at the time of transfection. FreeStyle™ CHO-S cells were cultured in CHOgro® Expression Medium or FreeStyle™ CHO Expression Medium and plated into non-treated 6-well plates (2ml/well) for transfection. Antibody levels were also analyzed from day 6 clarified supernatants using a human IgG ELISA (ZeptoMatrix). Error bars represent the standard deviation of triplicate technical replicates.

Achieve High Protein Yields Using TransIT-PRO® Transfection Kit in Suspension 293 Cells. Ten different secreted (non‐antibody) proteins were transiently expressed in FreeStyle™ 293‐F cells (Life Technologies) using TransIT‐PRO (1.5:1) or 293fectin™ (Life Technologies, 2:1) transfection reagents according to manufacturer's protocol. Cells were grown in FreeStyle™ 293 Expression Medium at transfected at a density of 1 x 10⁶ cells/ml. The scale of the transfection for each protein varied between 1‐6 L of culture. More experimental details here.

Data courtesy of a TransIT‐PRO pharmaceutical customer.