Cambio - Excellence in Molecular Biology

Transfection-EZ Biosystems

Transfection-EZ Biosystems: Broad Range Transfection Reagents

We offer a range of EZ Biosystems' general purpose transfection reagents for plasmid, siRNA, co-transfections, in vivo work and CRISPR.  These reagents are cost-effective and have been specially formulated for maximum transfection efficiency of a wide range of different cell types with minimal toxicity.

Why broad spectrum for all type of nucleic acids?

Avalanche® transfection reagents have the broad spectrum for the transfection of large plasmid, mRNA, siRNA, and/or other type of nucleic acids. This feature makes Avalanche® transfection reagents the best option for co-transfection of different type and/or size of nucleic acids.  The following are the reasons for the broad spectrum:

The key components of Avalanche® transfection reagents are mixtures of cationic lipids and cationic polymers, which contain all three types of amines: primary, secondary, and tertiary amines. The three types of amines have different positive charge strength, hence have different capabilities to combine with negative charged anions.

As anions, different types and sizes of nucleic acids have different densities and quantities of negative charges.

With optimized ratios of different cationic lipids and polymers, hence optimized ratios of the three different type of amines, together with optimized protocols, our Avalanche® mixtures have the optimized strength of positive charges that ensure them to efficiently combine with different type of nucleic acids of a variety of negative charge densities and quantities, and compact them into the right sizes of nanoparticles with the right densities of positive charge (zeta potential) on their surface. The optimized size and the right strength of zeta potential of the nanoparticles are the keys for the nanoparticles to interact efficiently with negatively charged cell surface, and trigger endocytosis process.

   

Avalanche-PRO Transfection Reagent

Avalanche-PRO Transfection Reagent

EZ Biosystems

Catalogue No.DescriptionPack SizePriceQty
EZT-PROT-1-15Avalanche-PRO Transfection Reagent 1.5ml £582.00 Quantity Add to Order
EZT-PROT-1-150Avalanche-PRO Transfection Reagent 15ml POA Quantity Add to Order

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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