Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

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TransIT-PRO® Transfection Kit

High, Reproducible Protein Yield in Suspension CHO and 293 Cells

Features

  • Ideal for biotherapeutic protein production
  • Achieve high protein yield in suspension CHO and 293 cells
  • Compatible with multiple CHO media formulations
  • Reproducible protein expression with minimal optimization

Description

  • Reach critical stages faster. Decrease time to produce usable protein by maximizing target protein yields through transient transfection. The TransIT-PRO® Transfection Kit uses animal origin free components designed for high and reproducible protein yield in suspension CHO and 293 derived cells. Since it is compatible with varied media formulations, the same media can be used for both transient and stable expression. The TransIT-PRO® outperforms linear PEI in protein yield, while providing a cost-effective alternative to FreeStyle™ MAX.
Sandwich ELISA after TransIT-PRO® transfection
Western blot after TransIT-PRO® transfection

Achieve high antibody titers using the TransIT-PRO® Transfection Kit in suspension CHO cells. Human IgG1 was produced by transient transfection using TransIT-PRO® and PRO Boost Reagent (1:1:1), 25 kDa linear PEI (6:1) or FreeStyle™ MAX (1:1) transfection reagents according to the manufacturers or published protocol (reagent:DNA ratio). Transfections were performed using 1 µg plasmid DNA per milliliter of culture and 0.5 x 106 cells/ml at the time of transfection. FreeStyle™ CHO-S cells were cultured in 20 ml of FreeStyle™ CHO Expression medium in 125 ml shake flasks. (A) Day 3, 5 and 7 supernatants were clarified and analyzed using a human IgG-Fc sandwich ELISA. Error bars represent the standard deviation of triplicate technical replicates, 25kDa linear PEI is duplicate technical replicates. (B) Day 7 supernatants were clarified and analyzed by Western blot. An IgG standard was included for quantification estimate (S1= 1.6 mg/l, S2= 3.2 mg/l, S3 = 6.3 mg/l).

Easy to use: compatible with multiple media formulations

Luciferase expression after TransIT-PRO® transfection
Human IgG1 quantitation after TransIT-PRO® transfection

TransIT-PRO® provides high performance across varied media formulations. FreeStyle™ CHO-S cells were adapted to five representative growth media including: BD Select™ CD1000 medium (Becton, Dickinson and Company, Franklin Lakes, NJ), BD Select™ CHO medium (Becton, Dickinson and Company, Franklin Lakes, NJ), FreeStyle™ CHO Expression medium (Life Technologies Corporation, Carlsbad, CA), ProCHO™5 medium (Lonza Inc., Allendale, NJ) and PowerCHO®2 CD medium (Lonza Inc., Allendale, NJ). Cells were transfected with a plasmid using the TransIT-PRO® and PRO Boost Reagent (1:1:1), 25 kDa linear PEI (4:1) (Polysciences, Warrington, PA), or FreeStyle™ MAX (1:1) (Life Technologies Corporation, Carlsbad, CA) transfection reagents according to the manufacturers or published protocol (reagent:DNA ratio). Transfections were performed in 24-well deep well shaker blocks using 1 µg plasmid DNA per milliliter of culture and 0.5 x 106 cells/ml at the time of transfection. (A) Luciferase expression was assessed 48 hours post-transfection using a conventional luciferase assay. Error bars represent the standard deviation of duplicate wells. (B) Human IgG1 was quantitated from day 5 clarified supernatants and analyzed by a human anti-Fc sandwich ELISA. Error bars represent the standard deviation of triplicate wells.

Scale-up: no problem

Anti-Fc sandwich ELISA after TransIT-PRO® transfection

Scaling of transient transfection using TransIT-PRO® Transfection Kit is linear from 4 to 400 milliliters. Human IgG1 was produced by transient transfection using the TransIT-PRO® Transfection Kit and a plasmid encoding a human IgG1 construct. DNA concentration was 1µg/ml of culture and a ratio of TransIT-PRO®: PRO Boost Reagent:DNA ratio of 1:0.5:1. Cells were plated at a density of 0.5 x 106 cells/ml at the time of transfection. CHO-S cells were cultured in BD Select™ CD1000 media using 4 ml per well of a 24-well deep well shake block, 40 ml in 125 ml Erlenmeyer shake flask, 40 ml in 125 ml 2 sidearm spinner flask and 400 ml in 500 ml 2 sidearm spinner flask. Day 3, 5 and 7 supernatants were clarified and analyzed by an anti-Fc sandwich ELISA. Error bars represent the standard deviation of triplicate technical replicates.

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
MIR5740 TransIT-PRO® Transfection Reagent 1 ml 373,00 PDF
MIR5750 TransIT-PRO® Transfection Reagent 10 ml 2973,00 PDF
MIR5700 TransIT-PRO® Transfection Kit (includes PRO Boost Reagent) 1 ml 399,00 PDF
MIR5760 TransIT-PRO® Transfection Kit (includes PRO Boost Reagent) 10 ml 3185,00 PDF
  • All prices are in EURO excl. VAT and shipping. Only available in selected countries.

  • * Sample Policy:
    MoBiTec together with Mirus considers sample requests for up to three different transfection products and one electroporation product per laboratory. Please include all samples to be tested in a single request.