Overview
Specifications
Product | FectoCHO® Expression system |
---|---|
Application | Protein / Antibody Production |
Kit components | FectoCHO® CD Expression Medium |
Cell types | Suspension CHO cells such as CHO-K1, CHO-S or ExpiCHO™-S |
Number of transfections | 1 mL of FectoPRO® Transfection reagent is sufficient to transfect 800 ml to more than 1 liter of FectoCHO® CD Expression Medium |
Storage | Store FectoPRO® and FectoCHO® CD Expression Medium at 5 °C ± 3°C. Do not freeze. |
Summary
The growing need in the biopharmaceutical market for various recombinant proteins and antibodies produced in a short time requires an efficient and flexible system. Transient expression is a commonly used process to face this demand but is unfortunately limited by transfection efficiency and inherent productivity, especially with CHO cells. To overcome this issue, we developed an advanced transient expression system, FectoCHO® Expression system, consisting in the synergistic association of a novel CHO chemically defined medium, FectoCHO® CD Expression medium and a powerful transfection reagent, FectoPRO®.
FectoCHO® CD Expression medium is optimised to facilitate adaptation and cultivation of various strains of CHO cells such as CHO-K1, CHO-S and ExpiCHO™-S : extensive sequential adaptation is not required. FectoPRO®-mediated transient transfection in this chemically defined medium is optimal and therefore leads to the highest protein and antibody yields using Transient Gene Expression; with up to 4 times higher protein yields than with the competitors’ systems.
Ordering information
Reference Number | Expression Medium | Transfection Reagent | Expression Booster |
---|---|---|---|
101000011 | FectoCHO® CD Medium 1 L | FectoPRO® 1 mL | 1mL |
101000003 | FectoCHO® CD Medium 6 x 1 L | - | - |
101000014 | - | FectoPRO® 1 mL | 1mL |
101000007 | - | FectoPRO® 10 mL | 10mL |
101000019 | - | FectoPRO® 100 mL | 4x10mL |
Description
Outstanding protein production yields in suspension CHO cells
The combination of a chemically defined medium optimized for transient transfection and protein expression, FectoCHO® CD Expression medium, and of a highly efficient transfection reagent, FectoPRO® leads to outstanding protein production yields in various CHO cells, such as CHO-S and CHO-K1 cells. (Fig. 1)
Fig. 1. Higher protein yields with FectoCHO® Expression system in CHO-S and CHO-K1 cells than with competitor’s Expression System. CHO-S cells were cultivated either in FectoCHO® CD medium or in FreeStyle™ CHO medium, and transfected with FectoPRO® reagent (0.5 µg DNA/mL) or FreeStyle™ MAX reagent (1.25 µg DNA/mL) following the recommended protocols. Suspension-adapted CHO-K1 cells were cultivated either in FectoCHO® CD Expression medium or in FreeStyle™ F17, then transfected with FectoPRO® (0.8 µg DNA/mL) or FreeStyle™ MAX (1.25 µg DNA/mL) following the recommended protocols. IgG3-Fc production was assayed over 7 days post-transfection using protein G Biosensors (fortéBIO® BLItz).
The ability of FectoCHO® Expression medium to maintain the cells in an ideal physiological state before, during and after transfection, coupled to the extremely high transfection efficiency of FectoPRO® leads to produce amazing protein and antibody yields using Transient Gene Expression. Indeed, FectoCHO® Expression System will boost your protein yields up to 4 times compared to competitors’ expression systems. (Fig. 2)
Fig. 2. The Synergistic combination of FectoPRO® Reagent and FectoCHO® CD Medium outperforms its main competitor in CHO-K1 (A) and CHO-S (B) cells. A. Suspension-adapted CHO-K1 cells were cultivated in either FectoCHO® CD Expression medium or FreeStyle™ F17, and transfected with FectoPRO® reagent (0.8 µg DNA/mL) or FreeStyle™ MAX reagent (1.25 µg DNA/mL) following the recommended protocols. IgG3-Fc production was assayed 7 days post-transfection using protein G Biosensors (fortéBIO® BLItz). B. CHO-S cells were cultivated either in FectoCHO® CD medium or in FreeStyle™ CHO medium, and transfected with FectoPRO® reagent (0.5 µg DNA/mL) or FreeStyle™ MAX reagent (1.25 µg DNA/mL) following the recommended protocols. IgG3-Fc production was assayed 7 days post-transfection using protein G Biosensors (fortéBIO® BLItz).
Ideal for high density cell systems such as ExpiCHO™-S cells
FectoCHO® Expression system is particularly well suited for high density cells, such as the ExpiCHO™-S cells. FectoCHO® CD medium is able to sustain growth and viability at very high cell density for up to 10 days, while ensuring a higher transfection efficiency with FectoPRO®, leading to very high protein yields in ExpiCHO™-S cells. (Fig. 3)
Fig. 3. FectoCHO® Expression system outperforms ExpiCHO™ Expression System in terms of protein production kinetics in ExpiCHO-S cells. ExpiCHO-S™ cells were either cultivated in FectoCHO® CD Expression Medium and transfected with FectoPRO® reagent following the recommended protocol, or in ExpiCHO™ Expression Medium and transfected following the recommended protocol with ExpiFectamine™ CHO transfection reagent. IgG3-Fc production was assayed over 10 days post-transfection using protein G Biosensors (fortéBIO® BLItz).
CHO cell adaptation
FectoCHO® CD Expression medium allows for CHO cells to reach high cell density, while maintaining an excellent viability with short doubling time. This medium has been specifically developed to allow a quick cell adaptation, whatever the CHO cell type used. (Fig. 4)
Fig. 4: ExpiCHO™-S adaptation in FectoCHO® CD Medium. ExpiCHO-S™ cells were thawed and maintained either in FectoCHO® CD Expression Medium or in ExpiCHO™ Expression Medium. Cell density and viability were measured using Trypan Blue exclusion. Similar results were obtained with CHO-S and CHO-K1 cells (data not shown).
Chemically defined kit components
FectoCHO® expression system is a 3 component kit optimized to work together:
- FectoCHO® CD Expression Medium, a chemically defined cell culture medium optimized for transient gene expression
- FectoPRO® Transfection Reagent to produce amazing protein yields using low DNA amount
- FectoPRO® Expression Booster that enhances gene expression to boost the protein production
Excellent lot to lot reproducibility
FectoCHO® Expression System components are manufactured and formulated using a highly controlled production process. This ensures highly reproducible results from experiment to experiment, hence data that can be easily repeated and protein productions with similar yields. (Fig. 5)
Fig. 5. FectoCHO® Expression system gives excellent lot-to-lot reproducibility. ExpiCHO™-S cells were cultivated in different FectoCHO® CD Expression Medium batches and transfected with FectoPRO® reagent following the recommended protocol. 7 days post-transfection, cell viability was measured using Trypan Blue exclusion. IgG3-Fc production was assayed using protein G Biosensors (fortéBIO® BLItz).
Application
FectoCHO® Expression system is perfectly suited for protein and antibody production using transient gene expression in various CHO cell lines, such as CHO-K1, CHO-S and ExpiCHO™-S cells. The chemically defined components of the kit have been optimized to provide synergistic effects and outstanding production yields.
📰 Read our Application note: “Improving expression and purification of mAb using FectoCHO® Expression System and MabXpure™.”
Quality
FectoCHO® Expression system is compliant with biomanufacturing guidelines. Indeed, FectoPRO®, FectoPRO® booster and FectoCHO® CD Expression medium are guaranteed free of animal-origin components, and are chemically defined. Systematic lot management and release testing is performed for each lot produced. Furthermore, each component of the FectoCHO® Expression System undergoes advanced quality controls such as 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.
Protocol
In order to download a product protocol or a certificate of analysis, please create an account on Polyplus ® 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!
Other files
Related blog posts
Bibliography
Cell Line | in vitro in vivo | Delivered Molecule | Reagent | Results & Citations | |
---|---|---|---|---|---|
- | in vitro | DNA | FectoCHO 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 |
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 | More details |
CHO-S | in vitro | DNA | FectoCHO Expression System, FectoPRO | Ehrencrona E. et al. (2021) J Biol Chem 11, The IgG Fc-binding protein FCGBP is secreted with all GDPH sequences cleaved, but maintained by inter-fragment disulfide bonds | More details |
CHO-S | in vitro | DNA | FectoCHO Expression System, FectoPRO | Ehrencrona E. et al. (2022) J Biol Chem 297, 100871 The IgGFc-binding protein FCGBP is secreted with all GDPH sequences cleaved but maintained by interfragment disulfide bonds | More details |
ExpiCHO-S, HEK-293T | in vitro | DNA | FectoCHO Expression System, FectoPRO | Kaku Y. et al. (2021) Cell Rep. 36, Resistance of SARS-CoV-2 variants to neutralization by antibodies induced in convalescent patients with COVID-19 | More details |
CHO-S, FreeStyle 293-F, Lec3.2.8.1 | in vitro | DNA | FectoCHO Expression System, FectoPRO | Samuelsson E. et al. (2022) ACS Infect Dis 8, 1883-1893 Sialic Acid and Fucose Residues on the SARS-CoV-2 Receptor-Binding Domain Modulate IgG Antibody Reactivity | More details |
CHO-S, Epithelial | in vitro | DNA | FectoPRO, FectoCHO Expression System | Good protein yield | Protocol available |