ExpiCHO Expression System
The Gibco ExpiCHO Expression System provides the highest protein yields possible in a transient system, and allows you to work in CHO cells from discovery to production.
FreeStyle™ CHO Expression Medium (Gibco™)
FreeStyle™ CHO Expression Medium is an animal origin–free, chemically defined, protein-free medium specifically developed to support the growth and transient transfection of CHO-S cells in suspension without adaptation.
This medium requires supplementation with 8 mM glutamine before use. Gibco® FreeStyle™ CHO Expression Medium features:
• Formulation without hypoxanthine and thymidine for use with or without dhfr systems
• Ability to support high-density suspension growth and transient transfection of CHO-S cells
• Animal origin–free, protein-free and chemically defined formulation
• Easy use as part of the FreeStyle™ MAX CHO Expression System for protein production
• Scalability in spinner flasks and bioreactors
Formulation without hypoxanthine and thymidine for use with or without dhfr systems
Gibco® FreeStyle™ CHO Expression Medium is made without hypoxanthine and thymidine for use in dihydrofolate reductase (dhfr) amplification systems. Non-dhfr systems require the addition of HT Supplement before use.
Ability to support high-density suspension growth and transient transfection of CHO-S cells
Gibco® FreeStyle™ CHO cells in suspension can be cultured directly into Gibco® FreeStyle™ CHO Expression Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into Gibco® FreeStyle™ CHO Expression Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations. Cells cultured in Gibco® FreeStyle™ CHO Expression Medium can be transfected using lipid-based transfection reagents, such as Freestyle™ MAX reagent.
Animal origin–free, protein-free and chemically defined formulation
Gibco® FreeStyle™ CHO Expression Medium is animal origin–free, protein-free, and chemically defined, allowing for easier purification of your protein of interest. Gibco® chemically defined media contain no proteins, hydrolysates, or components of unknown composition.
Easy use as part of the FreeStyle™ MAX CHO Expression System for protein production
The FreeStyle™ MAX CHO Expression System is a breakthrough technology for rapid (see figure) and high-yield mammalian protein production in 30 mL and 1 L batch sizes (see figure). The FreeStyle™ MAX CHO Expression System combines GIBCO® FreeStyle™ media, FreeStyle™ MAX reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293-F cells in one kit.
Scalability in spinner flasks and bioreactors
Protein production using Gibco® FreeStyle™ CHO Expression Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement Gibco® FreeStyle™ CHO Expression Medium with additional Pluronic® F-68 to avoid sheer stress in the culture.
cGMP manufacturing and quality system
Gibco® FreeStyle™ CHO Expression Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.
This medium requires supplementation with 8 mM glutamine before use. Gibco® FreeStyle™ CHO Expression Medium features:
• Formulation without hypoxanthine and thymidine for use with or without dhfr systems
• Ability to support high-density suspension growth and transient transfection of CHO-S cells
• Animal origin–free, protein-free and chemically defined formulation
• Easy use as part of the FreeStyle™ MAX CHO Expression System for protein production
• Scalability in spinner flasks and bioreactors
Formulation without hypoxanthine and thymidine for use with or without dhfr systems
Gibco® FreeStyle™ CHO Expression Medium is made without hypoxanthine and thymidine for use in dihydrofolate reductase (dhfr) amplification systems. Non-dhfr systems require the addition of HT Supplement before use.
Ability to support high-density suspension growth and transient transfection of CHO-S cells
Gibco® FreeStyle™ CHO cells in suspension can be cultured directly into Gibco® FreeStyle™ CHO Expression Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into Gibco® FreeStyle™ CHO Expression Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations. Cells cultured in Gibco® FreeStyle™ CHO Expression Medium can be transfected using lipid-based transfection reagents, such as Freestyle™ MAX reagent.
Animal origin–free, protein-free and chemically defined formulation
Gibco® FreeStyle™ CHO Expression Medium is animal origin–free, protein-free, and chemically defined, allowing for easier purification of your protein of interest. Gibco® chemically defined media contain no proteins, hydrolysates, or components of unknown composition.
Easy use as part of the FreeStyle™ MAX CHO Expression System for protein production
The FreeStyle™ MAX CHO Expression System is a breakthrough technology for rapid (see figure) and high-yield mammalian protein production in 30 mL and 1 L batch sizes (see figure). The FreeStyle™ MAX CHO Expression System combines GIBCO® FreeStyle™ media, FreeStyle™ MAX reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293-F cells in one kit.
Scalability in spinner flasks and bioreactors
Protein production using Gibco® FreeStyle™ CHO Expression Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement Gibco® FreeStyle™ CHO Expression Medium with additional Pluronic® F-68 to avoid sheer stress in the culture.
cGMP manufacturing and quality system
Gibco® FreeStyle™ CHO Expression Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.
FreeStyle™ CHO-S Cells (Gibco™)
Les cellules Freestyle™ CHO-S font partie du système d’expression FreeStyle™ MAXqui est une technologie particulièrement innovante permettant de générer rapidement et en grande quantité des protéines de mammifère. Cette lignée cellulaire est adaptée à la culture en suspension exempte de sérum à haut débit dans le milieu de culture d’expression FreeStyle™ CHO ; elle est capable de produire de hauts niveaux de protéines recombinantes.
FreeStyle™ MAX CHO Expression System (Gibco™)
The FreeStyle™ MAX System is a breakthrough technology for rapid and high-yield mammalian protein production. This new mammalian expression technology is available in two convenient formats. FreeStyle™ MAX Reagent is a novel cationic lipid-based reagent designed to transfect FreeStyle™ cells with high efficiency. The FreeStyle™ MAX Expression System combines GIBCO® FreeStyle™ Media, FreeStyle™ MAX Reagent, and either FreeStyle™ CHO-S cells or FreeStyle™ 293 cells. The only equipment required to use the FreeStyle™ MAX System is a CO2 incubator and a platform shaker with shake flasks or a spinner culture.
The FreeStyle™ MAX System provides:
• An easy, rapid protocol for transient transfection of CHO and 293 cells in suspension cultures (Figure 1)
• Functional proteins (up to milligram quantities) with all post-translational modifications produced in one week
• Simplified downstream purification of secreted proteins with FreeStyle™ serum-free medium
• Straightforward protocol for production scale-up
The FreeStyle™ MAX System can replace time-consuming stable cell line generation with large-scale functional protein production in one
week (Figure 2).
Contents and Storage:
Store FreeStyle™ cells in liquid nitrogen. Store FreeStyle™ MAX Reagent at +4°C. Store GIBCO® FreeStyle™ CHO Expression Medium, FreeStyle™ 293 Expression Medium, and OptiPRO™ SFM at +2-8°C in the dark. Store the pCMV SPORT at -20°C. Stability is guaranteed for 6 months under the recommended storage conditions. Please refer to the manual for additonal storage information.
The FreeStyle™ MAX System provides:
• An easy, rapid protocol for transient transfection of CHO and 293 cells in suspension cultures (Figure 1)
• Functional proteins (up to milligram quantities) with all post-translational modifications produced in one week
• Simplified downstream purification of secreted proteins with FreeStyle™ serum-free medium
• Straightforward protocol for production scale-up
The FreeStyle™ MAX System can replace time-consuming stable cell line generation with large-scale functional protein production in one
week (Figure 2).
Contents and Storage:
Store FreeStyle™ cells in liquid nitrogen. Store FreeStyle™ MAX Reagent at +4°C. Store GIBCO® FreeStyle™ CHO Expression Medium, FreeStyle™ 293 Expression Medium, and OptiPRO™ SFM at +2-8°C in the dark. Store the pCMV SPORT at -20°C. Stability is guaranteed for 6 months under the recommended storage conditions. Please refer to the manual for additonal storage information.
pcDNA™3.4 TOPO™ TA Cloning Kit (Invitrogen™)
Le vecteur ADNpc™ 3.4 TOPO® est un vecteur d’expression de cellules de mammifère constitutif conçu pour fournir des niveaux exceptionnellement hauts d’expression transgénique. Il peut être utilisé pour les cellules adaptées à la suspension, telles que les cellules Expi293F™, FreeStyle™ 293-F et FreeStyle™ CHO-S, pour l’expression des protéines transitoires. Le vecteur peut aussi être utilisé comme plasmide d’expression sélectionnable à la Geneticin® pour l’engineering de lignées cellulaires stables. L’accélérateur de CMV sur toute la longueur, optimisé, et d’autres éléments d’expression permettent généralement des niveaux d’expression plus élevés que les constructions d’expression basées sur l’ADNpc™ 3.1, 3.2 et 3.3.
Le vecteur ADNpc 3.4 TOPO® contient les éléments suivants :
• Longueur totale promoteur /accélérateur immédiat /précoce du cytomégalovirus (CMV) humain pour l’expression génétique de haut niveau dans une gamme étendue de cellules de mammifère
• WPRE (woodchuck posttranscriptional regulatory element = élément réglementaire post-transcriptionnel de la marmotte) en aval du site de clonage pour optimiser l’expression du produit de la transcription
• Site de clonage TOPO® pour un clonage rapide et efficace (>85 %) des produits de PCR amplifiés par Taq
• Gène de résistance à la néomycine pour la sélection de lignées cellulaires stables avec la généticine®
• Origine pUC pour une réplication en de nombreuses copies et maintenance du plasmide dans E. coli
Le kit de clonage d’ADNpc3.4™ TOPO® TA comprend un vecteur plasmidique adapté à TOPO®, des réactifs pour le clonage par PCR du gène de votre choix dans le vecteur, une amorce de séquençage pour confirmer l’orientation de l’insert et des cellules compétentes One Shot® TOP10.
Le vecteur ADNpc 3.4 TOPO® contient les éléments suivants :
• Longueur totale promoteur /accélérateur immédiat /précoce du cytomégalovirus (CMV) humain pour l’expression génétique de haut niveau dans une gamme étendue de cellules de mammifère
• WPRE (woodchuck posttranscriptional regulatory element = élément réglementaire post-transcriptionnel de la marmotte) en aval du site de clonage pour optimiser l’expression du produit de la transcription
• Site de clonage TOPO® pour un clonage rapide et efficace (>85 %) des produits de PCR amplifiés par Taq
• Gène de résistance à la néomycine pour la sélection de lignées cellulaires stables avec la généticine®
• Origine pUC pour une réplication en de nombreuses copies et maintenance du plasmide dans E. coli
Le kit de clonage d’ADNpc3.4™ TOPO® TA comprend un vecteur plasmidique adapté à TOPO®, des réactifs pour le clonage par PCR du gène de votre choix dans le vecteur, une amorce de séquençage pour confirmer l’orientation de l’insert et des cellules compétentes One Shot® TOP10.
FreeStyle™ F17 Expression Medium (Gibco™)
FreeStyle™ F17 Expression Medium is an animal origin–free, chemically defined, protein-free medium specifically developed to support the growth and transfection of 293-F and CHO cells in suspension without adaptation. This is a multi-platform medium compatible with a range of transient expression systems. The addition of 4–8 mM L-glutamine or GlutaMAX™ supplement is required before use. Gibco® FreeStyle™ F17 Expression Medium features:
• Little or no required adaptation
• Ability to support suspension growth and transfection of 293-F and CHO cells
• Animal origin–free, protein-free, and chemically defined formulation
• Scalability in spinner flasks and bioreactors
Little or no required adaptation
Gibco® 293-F, 293-H, FreeStyle™ 293-F, and CHO-S cells in suspension can be cultured directly into Gibco® FreeStyle™ F17 Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into Gibco® FreeStyle™ F17 Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations.
Ability to support suspension growth and transfection of 293-F and CHO cells
Gibco® FreeStyle™ F17 Medium provides equal or better performance compared to Gibco® FreeStyle™ 293 Expression Medium and Gibco® FreeStyle™ CHO Medium. Gibco® FreeStyle™ F17 Medium supports 293-F cells at a peak density of 6–7 × 106 viable cells per mL, and supports CHO-S cells at a peak density of 8–9 × 106 viable cells per mL. Cells cultured in Gibco® FreeStyle™ F17 can be transfected using a wide variety of transfection reagents, including lipid-based transfection reagents, such as 293fectin™ and Freestyle™ MAX Reagent. Protein expression typically exceeds 75–150 µg/mL IgG in 293-F cells.
Animal origin–free, protein-free, and chemically defined formulation
Gibco® FreeStyle™ F17 Medium is animal origin–free, protein-free, and chemically defined, allowing for easier purification of your protein of interest. Gibco® chemically defined media contain no proteins, hydrolysates, or components of unknown composition.
Scalability in spinner flasks and bioreactors
Protein production using Gibco® FreeStyle™ F17 Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement Gibco® FreeStyle™ F17 Medium with additional Pluronic® F-68 to avoid sheer stress in the culture.
cGMP manufacturing and quality system
Gibco® FreeStyle™ F17 Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.
• Little or no required adaptation
• Ability to support suspension growth and transfection of 293-F and CHO cells
• Animal origin–free, protein-free, and chemically defined formulation
• Scalability in spinner flasks and bioreactors
Little or no required adaptation
Gibco® 293-F, 293-H, FreeStyle™ 293-F, and CHO-S cells in suspension can be cultured directly into Gibco® FreeStyle™ F17 Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into Gibco® FreeStyle™ F17 Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations.
Ability to support suspension growth and transfection of 293-F and CHO cells
Gibco® FreeStyle™ F17 Medium provides equal or better performance compared to Gibco® FreeStyle™ 293 Expression Medium and Gibco® FreeStyle™ CHO Medium. Gibco® FreeStyle™ F17 Medium supports 293-F cells at a peak density of 6–7 × 106 viable cells per mL, and supports CHO-S cells at a peak density of 8–9 × 106 viable cells per mL. Cells cultured in Gibco® FreeStyle™ F17 can be transfected using a wide variety of transfection reagents, including lipid-based transfection reagents, such as 293fectin™ and Freestyle™ MAX Reagent. Protein expression typically exceeds 75–150 µg/mL IgG in 293-F cells.
Animal origin–free, protein-free, and chemically defined formulation
Gibco® FreeStyle™ F17 Medium is animal origin–free, protein-free, and chemically defined, allowing for easier purification of your protein of interest. Gibco® chemically defined media contain no proteins, hydrolysates, or components of unknown composition.
Scalability in spinner flasks and bioreactors
Protein production using Gibco® FreeStyle™ F17 Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement Gibco® FreeStyle™ F17 Medium with additional Pluronic® F-68 to avoid sheer stress in the culture.
cGMP manufacturing and quality system
Gibco® FreeStyle™ F17 Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.
FreeStyle™ MAX Reagent (Gibco™)
FreeStyle™ MAX Reagent is a proprietary, animal origin–free formulation for transfecting plasmid DNA into eukaryotic cells that can easily be scaled up to produce large amounts of recombinant proteins. FreeStyle™ MAX Reagent allows the highest expression levels and transfection rates with lowest cytotoxicity in bio-production applications, and is specifically formulated for use with:
• FreeStyle™ 293-F Cells (suspension human embryonal kidney cells, Cat. No. R790-07) in serum-free FreeStyle™ 293 Expression Medium (Cat. No. 12338-018)
• FreeStyle™ CHO-S Cells (suspension Chinese Hamster Ovary cells, Cat. No. R800-07) in serum-free FreeStyle™ CHO Expression Medium (Cat. No. 12651-014)
• DG44 Cells (DHFR– suspension CHO cells) in CD DG44 Medium with 8 mM L-glutamine and 18 mL/L 10% Pluronic® F-68 (Cat. No. A11000-01)
Product use
FreeStyle™ MAX Reagent is intended for use with the FreeStyle™ MAX 293 Expression System (Cat. No. K9000-10) and the FreeStyle™ MAX CHO Expression System (Cat. No. K9000-20).
• FreeStyle™ 293-F Cells (suspension human embryonal kidney cells, Cat. No. R790-07) in serum-free FreeStyle™ 293 Expression Medium (Cat. No. 12338-018)
• FreeStyle™ CHO-S Cells (suspension Chinese Hamster Ovary cells, Cat. No. R800-07) in serum-free FreeStyle™ CHO Expression Medium (Cat. No. 12651-014)
• DG44 Cells (DHFR– suspension CHO cells) in CD DG44 Medium with 8 mM L-glutamine and 18 mL/L 10% Pluronic® F-68 (Cat. No. A11000-01)
Product use
FreeStyle™ MAX Reagent is intended for use with the FreeStyle™ MAX 293 Expression System (Cat. No. K9000-10) and the FreeStyle™ MAX CHO Expression System (Cat. No. K9000-20).
