Avalanche®-PRO Transfection Reagent is a proprietary, animal origin-free formulation of lipids and polymers for transfecting plasmid DNA into eukaryotic cells that can easily be scaled up to produce large amounts of recombinant proteins. Avalanche®-PRO Transfection Reagent allows the highest expression levels and transfection rates with lowest cytotoxicity in bio-production applications, and is specifically formulated for use with suspension 293 and CHO derived cells, such as FreeStyle 293-F Cells (suspension human embryonal kidney cells, Life Technologies, Cat. no. R790-07), FreeStyle CHO-S Cells (suspension Chinese Hamster Ovary cells, Life Technologies, Cat. no. R800-07), and DG44 Cells (DHFR- suspension CHO cells).
Features:
- Highest recombinant protein yield
- Lesser plasmid DNA and Transfection Reagent needed
- Compatible with multiple CHO media formulations
- Proven Scalability
- Simplest transfection protocol
- Totally animal free
- Half cost and similar or higher performances per reacton as compared to FreeStyle™ Max Reagent.
In brief, Avalanche®-PRO Transfection Reagent is ideal for Biotherapeutic Protein Production
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| Figure 1. Lesser DNA and lesser transfection reagent are needed for Avalanche®-PRO Transfection Reagent to have similar performances as compared to FreeStyle Max Reagent. (A) FreeStyle 293-F Cells were cultured in 30 ml of FreeStyle 293 Expression Medium to a density of 1 x 106 cells/ml for the transfection of pCMV·SPROT beta-gal as plasmid DNA. For Avalanche®-PRO Reagent transfection, DNA and Transfection Reagent used are 30 µg and 24 µl respectively; For FreeStyle Max Reagent transfection, DNA and Transfection Reagent used are 37.5µg and 37.5 µl respectively according to the respective protocols. (B) FreeStyle™ CHO-S Cells were cultured in 30 ml of FreeStyle™ CHO Expression Medium to a density of 1 x 106 cells/ml for the transfection of human IgG plasmid DNA. For Avalanche®-PRO Reagent transfection, DNA and transfection reagent used were 30 µg and 30 µl respectively; For FreeStyle Max Reagent transfection, DNA and Transfection Reagent used are 37.5µg : 37.5 µl respectively according to the respective protocols. |
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| Figure 2. Avalanche®-PRO provides high performance across varied media formulations. FreeStyle™ CHO-S cells were adapted to 8 representative growth media including: ProCHO™5 medium (Lonza Inc.), PowerCHO 2 CD medium (Lonza Inc), HyCell* CHO Medium (Thermo Scientific), FreeStyle™ CHO Expression medium (Life Technologies), Cellvento™ CHO-100 (EMD Millipore ), BD Select™ CHO medium (BD Biosciences), BD Select™ CD1000 medium (BD Biosciences), and EX-CELL® CD CHO-2 Medium (Sigma-Aldrich). Cells were transfected with a human IgG plasmid DNA using Avalanche-PRO (reagent:DNA=1.0 µl:1.0 µg/ml culture), or FreeStyle Max (Life Technologies) (reagent:DNA=1.25 µl:1.25 µg/ml culture) transfection reagents according to the published protocol ). Transfections were performed in 24-well deep well shaker blocks using 1 x 106 cells/ml at the time of transfection. Human IgG was quantitated from day 6 clarified supernatants and. Error bars represent the standard deviation of triplicate wells. |
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| Figure 3. Avalanche®-PRO Transfection Reagent Scalability and Cost Saving as compared to FreeStyle Max Reagent. (A)Avalanche®-PRO transfections in culture volumes from 1 ml to 250 ml resulted in consistant Human IgG yield in terms of µg/ml culture. (B) and (C): While the transfection performance of Avalanche®-PRO Transfection Reagent is similar to that of FreeStyle MAX Reagent, the cost of Avalanche®-PRO needed per reacton is only half of the cost of FreeStyle MAX Reagent needed. |
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