NEW! CHOgro® High Yield Expression System

The most advanced system for transient transfection and protein production in suspension CHO cells

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MIR 62701 Kit

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The NEW CHOgro® High Yield Expression System is the most advanced and cost-effective transient transfection system for high-yield protein production in suspension CHO cells. Our second-generation system features the CHOgro® Titer Enhancer, which provides rapid, industry-leading protein yields.

  • High Yield - Reach higher antibody titers in seven days - faster than the ExpiCHO Expression System
  • Simple Workflow - Same-day transfection, enhancer addition and temperature shift
  • Worry Free - No commercial license required; animal origin-free

Figures and Data

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The CHOgro High Yield Expression System Outperforms the ExpiCHO Expression System Using Multiple Antibody Constructs
The CHOgro® High Yield Expression System Outperforms the ExpiCHO Expression System Using Multiple Antibody Constructs. Six different IgG1 antibody constructs, including five therapeutically relevant constructs synthesized in the same vector backbone were produced by transient transfection using either the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt) and 1 µg plasmid DNA per milliliter of culture in FreeStyle™ CHO-S cells (ThermoFisher Scientific) cultured in CHOgro® Expression Medium (Mirus Bio) at a cell density of 4 x 106 cells/ml, or the Expifectamine CHO Transfection Kit (ThermoFisher Scientific) at a 3.2:1 reagent-to-DNA (vol:wt) and 1 µg plasmid DNA per milliliter of culture in ExpiCHO-S cells (ThermoFisher Scientific) cultured in ExpiCHO Expression Medium (ThermoFisher Scientific) at a cell density of 6 x 106 cells/ml. All cells were split into 125 ml Optimum Growth flask (25 ml/flask, Thomson Instrument Company) at the time of transfection. The CHOgro® Titer Enhancer was added immediately post-addition of transfection complexes and the culture was incubated at 32 °C shaking until harvest for the CHOgro® High Yield Expression System. The Max titer protocol was followed for the ExpiCHO Expression System (ThermoFisher Scientific): Expifectamine CHO Enhancer and Feed (ThermoFisher Scientific) were added at 24 hours post-transfection and cultures were shifted to 32 °C. At Day 5 a second volume of the ExpiCHO Feed (ThermoFisher Scientific) was added to the flask. Day 7 and 14 supernatants were analyzed using a standard sandwich human IgG ELISA. Error bars represent the standard deviation of triplicate technical replicates.
 
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CHOgro Titer Enhancer Does Not Adversely Affect Cell Growth and Viability Post-transfection

CHOgro® Titer Enhancer Does Not Adversely Affect Cell Growth and Viability Post-transfection. Triplicate flasks of FreeStyle™ CHO-S cells adapted to CHOgro® Expression Medium were transiently transfected with the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt) and 1 µg plasmid DNA per milliliter of culture at 4 x 106 cells/ml in 125 ml Optimum Growth flasks (25 ml/flask, Thomson Instrument Company). As indicated, CHOgro® Titer Enhancer was added and all of the cultures were shifted to 32 °C immediately post-addition of the transfection complexes to the culture. Cell counts (solid line) and viability (propidium iodide staining, dotted line) were measured daily post-transfection using a Guava® easyCyte™ 5HT flow cytometer (EMD Millipore). 

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Titer Enhancement is Observed Using Multiple Fc-fusion Constructs
Titer Enhancement is Observed Using Multiple Fc-fusion Constructs. Two different Fc-fusion constructs were transiently transfected using the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt) and 1 µg plasmid DNA per milliliter of culture. FreeStyle™ CHO-S (ThermoFisher Scientific) cells were cultured in CHOgro® Expression Medium (Mirus Bio) and plated at 4 x 106 cells/ml at the time of transfection in non-treated 6-well plates (2ml/well). As indicated, CHOgro® Titer Enhancer was added and the culture was shifted to 32 °C immediately post-addition of the transfection complexes to the culture. Day 10 supernatants were analyzed using a standard sandwich human IgG ELISA. Error bars represent the standard deviation of triplicate technical replicates.
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Third Party Validation of Increased Titers Obtained with CHOgro Titer Enhancer
Validation of Increased Titers Obtained with CHOgro® Titer Enhancer. An Fc-fusion protein was produced by transient transfection of CHO-S cells in complete CHOgro® Medium (Mirus Bio) using TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt). As indicated, CHOgro® Titer Enhancer (Mirus Bio) was added and the culture was shifted to 32˚C following transfection. Yields represent final affinity-purified protein from 0.5 L culture, harvested at day 10 post-transfection. Data courtesy of Lytic Solutions, LLC.
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TransIT-PRO + CHOgro Titer Enhancer Outperforms PEImax and linear 25 kDa PEI Transfection Reagents
TransIT-PRO® + CHOgro® Titer Enhancer Outperforms PEImax and linear 25 kDa PEI Transfection Reagents. An IgG1 antibody construct was produced by transient transfection using either the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt), PEImax (4:1, Polysciences), or linear 25 kDa PEI (6:1, Polysciences) and 1 µg plasmid DNA per milliliter of culture in FreeStyle™ CHO-S cells (ThermoFisher Scientific) cultured in CHOgro® Expression Medium (Mirus Bio) at a cell density of 4 x 106 cells/ml. CHOgro® Titer Enhancer (Mirus Bio) was added immediately post-addition of transfection complexes and cultures were incubated at 32 °C with shaking until harvest for all reagents. Day 14 supernatants were analyzed using a standard sandwich human IgG ELISA. Error bars represent the standard deviation of triplicate technical replicates.

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