Overview

Specifications

Reagent

FectoPRO®

Molecule delivered

DNA

Applications

Bioproduction

Cell types

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

Storage

5°C ± 3°C, stable for 3 years from the manufacturing date when stored appropriately

Provided with

FectoPRO® Booster


Summary

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.

Discover our New Kit: FectoCHO® Expression system

image-de-note-de-musique-7 Webinar: A Different Approach to Achieve a Higher Transfection Efficiency

📰 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®

Ordering information

Reference NumberProductTransfection ReagentExpression BoosterExpression Medium
116-001FectoPRO® Reagent1 mL1 mL-
116-010FectoPRO® Reagent10 mL10 mL-
116-100FectoPRO® Reagent100 mL100 mL-
716-06LFectoCHO® CD Medium--6 x 1 L
716-01LKITFectoCHO® Expression System1 mL1 mL1 L

Bulk quantities are available upon request.

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Description

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).

FectoPRO - antibody yields CHO cells

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).

Discover our Kit dedicated to cho cells: FectoCHO® Expression system

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).

FectoPRO - Microscopie

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.

FectoPRO - CHO cells

FectoPRO - HEK-293F cells

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).

Discover our Kit dedicated to cho cells: FectoCHO® Expression system

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.

FectoPRO - Table comparison competitor

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.).

FectoPRO - Production + viability in HEK-293

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).

Easily scalable

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.).

FectoPRO - CHO viability FectoPRO - CHO protein production

FectoPRO - Scalability

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).

FectoPRO - Reproductibility batch to batch

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).

FectoPRO - protein production

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).

FectoPRO - Table media CHO

Table 2. Protein yields using FectoPRO® in different culture media for CHO cells.

FectoPRO - Table media HEK

Table 3. Protein yields using FectoPRO® in different culture media for HEK-293 cells.

Discover our Kit dedicated to CHO cells: FectoCHO® Expression system

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.

FAQ

If you have any questions regarding FectoPRO®, please visit our dedicated Frequently asked questions or contact us.

Applications

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:

Read more…

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.

Read more…

Quality

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.

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.

Testimonials

"The solution we identified was to eliminate the need for the ExpiCHO-S cell Enhancer component by using instead a different transfection medium culture system, FectoPRO® with Booster, with the ExpiCHO-S cells. We also note that the FectoPRO® with Booster system is ~40% cheaper than its ExpiFectamine CHO with Feed and Enhancer counterpart. This methodology change would therefore also provide cost savings."
doi: 10.1371/journal.pone.0215106.
 Cupo et al, 2019 
  
"In our hands, FectoPRO® gives the highest protein production yields we ever reached using transient transfection. Our company offers custom services including protein production in mammalian cells. Being able to produce a lot more protein without revisiting our expression system opens a whole new range of possibilities for our antibody and protein production services."
 Philippe F., ProteoGenix SAS, France 
  
“I have to admit I had low expectations for FectoPRO®. Perhaps a modest improvement in transfection … an extra 5-10%. J. did her first experiment yesterday and analyzed it today and the result was remarkable. I’m shocked by both the efficiency of transfection and the brightness of the GFP.”
 Thomas C., Biotech, Cambridge, MA, United States 
  
“FectoPRO® yields transient expression titers in HEK, matching or exceeding those from other suppliers, but at a fraction of the price.”
 James L., Biotech company, United States 
  
“2 x the yield for half the amount of transfection reagent = 4 x less cost. It is also very easy to use.”
 Jean-Philippe J., Hospital for Sick Children, Canada 
  

Bibliography

Order by :  
Found 42 results :
Cell Linein vitro
in vivo
Delivered MoleculeReagentResults & Citations
HEK-293FT, HEK-293Sin vitroDNAFectoPRO
Moyo, T. et al. (2018)

J Virol ,
Molecular basis of unusually high neutralization resistance in tier 3 HIV-1 strain 253-11
More details
ExpiCHO-Sin vitroDNAFectoPRO
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
More details
Expi293Fin vitroDNAFectoPRO
Brigger D. et al. (2020)

Allergy ,
Accuracy of serological testing for SARS-CoV-2 antibodies: first results of a large mixed-method evaluation study
More details
Expi293Fin vitroDNAFectoPRO
Elmer BM. et al. (2019)

PLoS One 14, e0226245
Gene delivery of a modified antibody to A
More details
-in vitroDNAFectoPRO
Zhang X. et al. (2021)

Commun Biol 4, 198
pH-dependent and dynamic interactions of cystatin C with heparan sulfate
More details
Expi293Fin vitroDNAFectoPRO
Darricarrere, N. et al. (2018)

J Virol ,
Development of a Pan-H1 Influenza Vaccine
More details
Expi293F, HEK-293Tin vitroDNAFectoPRO, jetPRIME
Gardner, M. R. et al. (2019)

Science Translational Medicine ,
AAV-delivered eCD4-Ig protects rhesus macaques from high-dose SIVmac239 challenges
More details
FreeStyle 293-Fin vitroDNAFectoPRO
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
More details
FreeStyle 293-Fin vitroDNAFectoPRO
Wang R. et al. (2019)

Front Cell Infect Microbiol 9, 417
A Murine Monoclonal Antibody With Potent Neutralization Ability Against Human Adenovirus 7
More details
-in vitroDNAFectoCHO Expression System
Ringe RP. et al. (2020)

J Virol 94, e01883-19
Neutralizing Antibody Induction by HIV-1 Envelope Glycoprotein SOSIP Trimers on Iron Oxide Nanoparticles May Be Impaired by Mannose Binding Lectin
More details