EZT-PROT-1 Protocol.
Description
Transfection reagent 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 reaction 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 ®g and 24 ®l respectively; For FreeStyle Max Reagent transfection, DNA and Transfection Reagent used are 37.5®g and 37.5 ®l 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 ®g and 30 ®l respectively; For FreeStyle Max Reagent transfection, DNA and Transfection Reagent used are 37.5®g : 37.5 ®l 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 ®l:1.0 ®g/ml culture), or FreeStyle Max (Life Technologies) (reagent:DNA=1.25 ®l:1.25 ®g/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 ®g/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.
Additional Information
| Weight | 0.5 lbs |
|---|---|
| Product Sizes |
1.5 ml, 15 ml |
| Subcategories |
For Protein Production Transfection |
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