Protein production in 293 & CHO derived cells: TransIT-PRO

The TransIT-PRO® transfection kit from Mirus Bio was specifically developed for high yield mammalian protein production in suspension HEK 293 and CHO derived cells. Combining a high efficiency, reproducible, animal origin-free DNA transfection reagent (TransIT-PRO) and an optional boost reagent that enhances expression in certain media formulations (PRO boost), the TransIT-PRO transfection kit achieves exceptional protein yield whilst eliminating the need for a culture medium change post-transfection.

TransIT-PRO is now is a part of a complete high yield CHOgro Expression System developed to achieve high yields in suspension CHO cells.

Benefits of using the TransIT-PRO Kit
• High protein yield even in CHO-S and Expi293F™ cells
• Simple workflow
• Reproducible protein expression with minimal optimisation
• Produce usable protein faster
• Compatible with multiple CHO media formulations
• Animal origin-free
• PRO Boost Reagent enhances expression in certain media formulations
• Compatible with CHOgro® expression media for highest yield in suspension CHO cells

High protein yields achieved using TransIT-PRO Transfection Kit in suspension 293 cells

Ten different secreted (nonantibody) 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.

TransIT-PRO yields multifoldincreases in antibody titre 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 millilitre 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 nontreated 6-well plates (2ml/well) for transfection. Antibody levels were also analysed from day six clarified supernatants using a human IgG ELISA (ZeptoMatrix). Error bars represent the standard deviation of triplicate technical replicates.

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 millilitre 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.

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Material available for download
TransIT-PRO Transfection Kit (includes PRO Boost Reagent) protocol
TransIT-PRO Transfection Reagent protocol
TransIT-PRO Transfection Kit (includes PRO Boost Reagent) quick ref protocol
TransIT-PRO Transfection Reagent quick ref protocol
TransIT-PRO Transfection Kit (includes PRO Boost Reagent) SDS
TransIT-PRO Transfection Reagent SDS
Selecting a transfection reagent for large scale protein production in suspension 293 cell types

TransIT-PRO Kit citation list

FreeStyle™ and 293fectin™ are trademarks of Life Technologies
BD Select™ is a trademark of Becton, Dickinson and Company
ProCHO™ and PowerCHO® are trademarks of Lonza Inc.