产品资料

Cambio/Avalanche-PRO Transfection Reagent/EZT-PROT-1-150

Description

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

Data:

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.

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.

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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Cambio/Avalanche-PRO Transfection Reagent/EZT-PROT-1-150

Description