FectoPRO®

High yields in suspension CHO & HEK-293

  • Amazing protein and antibody production yields
  • Cost-effective Transient Gene Expression using lower plasmid DNA amount
  • Compatible with various mammalian expression media & cells systems
  • Compliant with biomanufacturing guidelines

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

4°C, stable for 1 to 2 years when stored appropriately

Provided with

FectoPRO® Booster

Summary

For the first time, achieve superior protein yields using transient transfection and gene expression.

FectoPRO® Transfection kit 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.

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

File icone GEN article: Upping CHO and HEK-293 Cell Productivity

Ordering information

Catalog NumberVolume of FectoPRO® ReagentVolume of FectoPRO® Booster
116-0011 ml1 ml
116-01010 ml10 ml
116-0404 x 10 ml4 x 10 ml
704-10-10 ml

Bulk quantities are available upon request.

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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).

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).

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 106cells/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 (A) and IgG3-Fc production was assayed by protein G affinity quantification (HPLC) (B) and Western Blot (C). Standard (C) 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.

FreeStyleTM, CD FortiCHOTM, CD OptiCHOTM, Expi293TM and CD293 are trademarks of GibcoTM.
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.

FectoPRO is also perfectly suited for virus production.

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…

Batonick, M., Kiss, M.M., Fuller, E.P., Magadan, C.M., Holland, E.G., Zhao, Q., Wang, D., Kay, B.K., Weiner, M.P. (2016). pMINERVA: A donor-acceptor system for the in vivo recombineering of scFv into IgG molecules, J Immunol Methods, doi: 10.1016/j.jim.2016.02.003.

Ferreira de Freitas, R., Eram, M. S., Smil, D., Szewczyk, M. M., Kennedy, S., Brown, P. J., Santhakumar, V., Barsyte-Lovejoy, D., Arrowsmith, C. H., Vedadi, M., Schapira, M. (2016). Discovery of a Potent and Selective Coactivator Associated Arginine Methyltransferase 1 (CARM1) Inhibitor by Virtual Screening, J Med Chem, doi: 10.1021/acs.jmedchem.6b00668.

Li, M., Yang, S., Xu, D. (2016). Heparan Sulfate Regulates the Structure and Function of Osteoprotegerin in Osteoclastogenesis, J Biol Chem, doi: 10.1074/jbc.M116.751974.

“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

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.