Suspension CHO, HEK-293, and derivatives
|Number of transfections|
1 mL of FectoPRO® is sufficient to transfect 800 ml to more than 1 liter of cell culture
Store FectoPRO® at 5 °C ± 3°C.
For the first time, achieve superior protein yields using transient transfection and gene expression.
FectoPRO® Transfection reagent is specifically designed for enhanced Transient Gene Expression (TGE) in suspension CHO and HEK-293 cells in various serum-free media, using low DNA amount (< 1 µg/ml of cell culture).
FectoPRO®-mediated transfection process is easily scalable from a few ml to several liters of cell culture, ensuring robust reproducible protein production. Increased productivity and low DNA amount requirement make FectoPRO® a cost-effective solution for bioproduction processes.
📰 GEN article: Upping CHO and HEK-293 Cell Productivity
📰 Cell Culture Dish Article with Protéogénix: Improved Protein yields in CHO and HEK293 cells using a dedicated transfection reagent: FectoPRO®
|Reference Number||Transfection Reagent||Expression Booster||Expression Medium|
|101000014||FectoPRO® 1 mL||1 mL|
|101000007||FectoPRO® 10 mL||10 mL|
|101000019||FectoPRO® 100 mL||100 mL|
|101000011||FectoPRO® 1 mL||1 mL||FectoCHO® CD Medium 1|
|101000003||x||x||FectoCHO® CD Medium 6 x 1 L|
Amazing protein yields
FectoPRO® enables amazingly high protein and antibody yields in both CHO and HEK-293 cells grown in suspension in serum-free media (Fig 1).
Fig. 1. FectoPRO® gives amazing antibody production yields in suspension CHO cells. Cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyle™ CHO Expression Medium and transfected with FectoPRO® (0.8 µg DNA/ml) or FreeStyle™ MAX Reagent (1.25 µg DNA/ml) following recommended protocols for the respective reagents. Recombinant mouse IgG production yield was assayed 1 to 6 days after transfection using protein G biosensor (fortéBIO® octet RED96 system). FreeStyle™ is a trademark of Life Technologies™ Corporation. Data kindly provided by ProteoGenix SAS (France).
FectoPRO® largely outperforms other transfection reagents such as linear PEI commonly used in Bioproduction processes in terms of transfection efficiency (Fig. 2), leading to superior protein production yields (Fig. 3).
Fig. 2. FectoPRO® gives high transfection efficiency in suspension CHO cells. FreeStyle™ CHO-S cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyle™ CHO Expression Medium and transfected with FectoPRO® + FectoPRO® Booster or FreeStyle™ MAX Reagent using 0.4 µg/ml and 1.25 µg/ml of a GFP expressing plasmid, respectively. GFP expression was assayed 24 h after transfection using fluorescence microscopy.
Fig. 3. FectoPRO® gives superior protein production yields in suspension CHO and HEK-293 cells. FreeStyle™ CHO-S and HEK-293F cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyle™ CHO or 293 Expression Medium and transfected with FectoPRO® + FectoPRO® Booster (0.4 µg DNA/ml), FreeStyle™ MAX Reagent (1.25 µg DNA/ml) or PEIpro® (1 µg DNA/ml) following recommended protocols for the respective reagents. IgG3-Fc (34 kDa) production was assayed 72 h (CHO cells) and 120 h (HEK-293 cells) after transfection by protein G affinity quantification (HPLC) and Western Blot. Standard corresponds to purified IgG3-Fc (34 kDa).
Cost-effective: uses low plasmid DNA amounts
High quality plasmid DNA preparation can be very costly, especially when large DNA amounts and many plasmids are required for large-scale protein production. FectoPRO® saves 40 to 60% on DNA cost by using 0.4 to 0.6 µg DNA / 106 cells / ml of cell culture; hence making this kit an affordable and economical solution for Bioproduction processes.
Table 1. Amounts of DNA and volumes of reagent (FectoPRO® and competitor) for transfection according to manufacturers’ recommendations.
Sustained protein production
The duration of protein production processes has increased in the past few years, and most of the protein production processes now last 1-2 weeks to obtain optimal amount of produced protein. FectoPRO® has been developed to ensure sustained protein production over several days, while maintaining high cell viability for longer lasting protein production (Fig. 4.).
Fig. 4. FectoPRO® gives sustained protein production in suspension cells. FreeStyle™ HEK-293F cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyle™ 293 Expression Medium and transfected with FectoPRO® + FectoPRO® Booster (0.5 µg DNA/ml) following the standard protocol. IgG3-Fc production was assayed after different timepoints by protein G affinity quantification (HPLC). Similar results are obtained with FreeStyle™ CHO-S (data not shown).
Streamline your bioproduction process with a robust protocol that is easily adapted to various culture vessels. FectoPRO®-mediated transfection can be scaled linearly from 30 ml up to liters of cell culture without protocol adaptation (Fig. 5.).
Fig. 5. FectoPRO® is easily scalable and reproducible in CHO cells. FreeStyle™ CHO-S cells were seeded at 1 x 106 cells/ml in 30 ml, 100 ml, 300 ml or 1 L of FreeStyle™ CHO Expression Medium and transfected with FectoPRO® + FectoPRO® Booster (0.5 µg DNA/ml). At day 1 to 6, cell viability was assayed using Trypan blue exclusion assay and IgG3-Fc production was assayed by protein G affinity quantification (HPLC) and Western Blot. Standard corresponds to purified IgG3-Fc (34 kDa).
Reproducible production yields
FectoPRO® is manufactured and formulated using a highly controlled production process. This ensures highly reproducible results from experiment to experiment. Moreover, in order to meet current regulatory guidelines, FectoPRO® is released using advanced quality controls including a specification for protein productivity that guarantees excellent lot-to-lot consistency (Fig. 6).
Fig. 6. FectoPRO® gives excellent lot-to-lot reproducibility. Suspension CHO cells were seeded at 1 x 106 cells/ml in 30 ml of CHO medium and transfected using 3 different lots of FectoPRO® (CCIII 078, CCIII 104 or CCIII 106) following the standard protocol. 72 h after transfection, IgG3-Fc production was assayed by protein G affinity quantification. Similar results are obtained on suspension HEK-293 cells (data not shown).
Compatible with mammalian expression systems
Unlike other commercially available reagents, FectoPRO® transfection kit is suitable for both CHO and HEK-293 cells (Fig. 2 and 3). The reagent is compatible with various media formulations; hence lab implementation is easy and straightforward (Fig. 7, Table 2).
Fig. 7. FectoPRO® gives superior protein production yields in suspension CHO cells grown in different media. CHO cells were seeded at 1 x 106 cells/ml in 30 ml of FreeStyle™ CHO Expression Medium, CD CHO Medium or Pro-CHO™ 4 Medium and transfected with FectoPRO® + FectoPRO® Booster or FreeStyle™ MAX Reagent following recommended protocols for the respective reagents. IgG3-Fc production was assayed 72 h after transfection by protein G affinity quantification (HPLC).
Table 2. Protein yields using FectoPRO® in different culture media for CHO cells.
Table 3. Protein yields using FectoPRO® in different culture media for HEK-293 cells.
FreeStyleTM, CD FortiCHOTM, CD OptiCHOTM, Expi293TM, ExpiCHOTM and CD293 are trademarks of Gibco®.
Pro-CHOTM, PowerCHOTM and Pro293TM are trademarks of LonzaTM Group.
HyCloneTM, HyCellTM and TransFxTM are trademarks of GE HealthcareTM.
BalanCD® and TransfectoryTM are trademarks of Irvine ScientificTM.
Transient protein production
Protein production consists in overexpressing a recombinant protein or peptide in a cell line at small / medium to large scale. The biomolecules produced can be used for research purposes like structural characterization all the way down to therapeutic or medical applications.
📰 Available Application Notes:
- Optimization of Glycoprotein Expression by Transient Transfection in HEK 293F/S suspension cells
- Superior protein yields in suspension CHO cells using FectoPRO®-mediated transient transfection in CELLSTAR® CELLreactor™
- Improved Protein yields in CHO cells using a novel transfection reagent for TGE: FectoPRO®
- Improving expression and purification of mAb using FectoCHO® Expression System and MabXpure™
High density cell systems for protein production using FectoPRO®
Recombinant protein production for therapy and diagnostic applications is mainly achieved using HEK-293 and CHO cells cultivated in suspension cultures without serum. The latter has been removed from raw materials for various reasons (costs, downstream purification steps, reproducibility…) while culture in suspension has been implemented to increase productivity. Improvements in culture processes and cell line engineering led to the generation of high-density mammalian transient protein expression systems.
FectoPRO® is compliant with biomanufacturing guidelines. Indeed, FectoPRO® is guaranteed free of animal-origin components, and is chemically defined. Systematic lot management and release testing is performed for each lot produced. Furthermore FectoPRO® transfection kit undergoes advanced quality controls: protein productivity, cell viability, and complete sterility. Each product is provided with a Certificate of Analysis.
In addition, Polyplus-transfection® is ISO 9001 Quality Management System accredited since 2002; this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.
In order to download a product protocol or a certificate of analysis, please create an account on Polyplus-transfection® Portal.
Why would you need to create an account?
In this personal area you will have access to:
- Product Protocols
- Certificates of Analysis
- Exclusive webinars/articles
- And surprise features!
Related blog posts
|Cell Line||in vitro|
|Delivered Molecule||Reagent||Results & Citations|
Cupo, A. et al. (2019)
PLoS One 14, e0215106
Optimizing the production and affinity purification of HIV-1 envelope glycoprotein SOSIP trimers from transiently transfected CHO cells
Brigger D. et al. (2020)
Accuracy of serological testing for SARS-CoV-2 antibodies: first results of a large mixed-method evaluation study
Elmer BM. et al. (2019)
PLoS One 14, e0226245
Gene delivery of a modified antibody to A
|FreeStyle 293-F||in vitro||DNA||FectoPRO|
Zhang X. et al. (2021)
Commun Biol 4, 198
pH-dependent and dynamic interactions of cystatin C with heparan sulfate
Tanaka S. et al. (2021)
Sci Rep 11(1),
An ACE2 Triple Decoy that neutralizes SARS-CoV-2 shows enhanced affinity for virus variants
|ExpiCHO-S||in vitro||DNA||FectoCHO Expression System, FectoPRO|
Derking R. et al. (2021 )
Cell Rep 35, 108933
Enhancing glycan occupancy of soluble HIV-1 envelope trimers to mimic the native viral spike
Esmail S. et al. (2021)
Cell Rep Method 1, 100011
Rapid and accurate agglutination-based testing for SARS-CoV-2 antibodies
|HEK-293FT, HEK-293S||in vitro||DNA||FectoPRO|
Moyo, T. et al. (2018)
J Virol ,
Molecular basis of unusually high neutralization resistance in tier 3 HIV-1 strain 253-11
|Expi293F, HEK-293T||in vitro||DNA||FectoPRO, jetPRIME|
Gardner, M. R. et al. (2019)
Science Translational Medicine ,
AAV-delivered eCD4-Ig protects rhesus macaques from high-dose SIVmac239 challenges
|FreeStyle 293-F||in vitro||DNA||FectoPRO|
Koralewski R. et al. (2020)
J Med Chem ,
Discovery of OATD-01, a First-in-Class Chitinase Inhibitor as Potential New Therapeutics for Idiopathic Pulmonary Fibrosis