Gibco Freedom CHO-S Kit Frequently Asked Questions

Volumetric titers of >1 g/L in simple fed batch, and >3 g/L in fed batch with complex feeds have been obtained with the Freedom CHO-S Kit.

It is possible to go from transfection to lead clone in as little as four months with the Freedom CHO-S Kit.

Yes. After execution and payment for a Commercial Production License, a Cell Line Documentation Package (>200 pages, and specific to the lot of cells purchased), is provided which includes the entire lineage history of the cells starting at receipt of the initial vial at Life Technologies through delivery to the purchaser, as well as all testing reports.

CHO-S clones producing a model IgG protein were assessed for stability by comparing volumetric productivity for over 60 generations. Four out of the five clones that were tested maintained expression levels over 60 generations. Expression levels were considered stable if the titer at the end of the stability assessment (at generation 60) was at least 70% of the titer observed at the beginning of the stability study.

Yes. Freedom CHO-S cells are intended (with the purchase of a commercial license) for the development of biologics. They have been produced in a cGMP animal origin-free environment, and have been scaled up in a commercial setting to produce products for clinical testing.

The kit includes components for transfection, expression, clone creation, and stable cell line selection. These include Gibco CD FortiCHO medium, Gibco OptiPRO SFM, Gibco FreeStyle MAX Reagent, Gibco CHO-S cells (cGMP banked), One Shot TOP10 Chemically Competent E. coli, and the Gibco Freedom pCHO 1.0 Expression Vector.

A certificate of analysis (COA) demonstrating suitability for research use is available to everyone who purchases Freedom CHO-S cells. A full documentation package, including the cell line's lineage, history, and cGMP testing results, is offered in conjunction with a commercial use license.

Freedom pCHO 1.0 can be used to express 1 or 2 subunits and has both DHFR and puromycin resistance as selection markers. Note: kanamycin resistance is used to select for the vector in bacteria.

Puromycin in combination with methotrexate (MTX) is used to select for stable pools following transfection with the Freedom pCHO 1.0 vector.

Gibco Dynamis medium and Gibco EfficientFeed C+ 2X Supplement are recommended. Dynamis medium is specifically designed to offer the highest batch and fed-batch culture performance and yield with recombinant CHO cells in a chemically defined environment. Both products are available in both a liquid and an Advanced Granulation Technology, or AGT, format.

Limiting dilution cloning is recommended with the Freedom CHO-S Kit. An evaluation of cloning in semi-solid medium in combination with ClonePix™ FL is underway.

FreeStyle MAX Reagent is included in the Freedom CHO-S Kit as the preferred transfection method. CHO-S cells are also compatible with the Neon Transfection System (optimization of instrument settings is required).

Only two transfections are recommended.

Only two transfections are recommended.

In terms of scaling-up from shake flask into a bioreactor, initial experiments indicate that antibody-expressing CHO-S clones readily scale-up into bioreactor vessels while maintaining growth and titer performance.

A commercial use license is required prior to the sale of expressed proteins, or when an IND submission is being prepared for first in human clinical studies.

No. Only a one-time license fee is required, and it covers the commercialization of an unlimited number of molecules using the Freedom CHO-S cell line.

No. Only a one-time license fee is required, and it covers the commercialization of an unlimited number of molecules using the Freedom CHO-S cell line.

Yes. Our Field Application Scientists provide technical support.

Yes, we offer limited cell line development services. Please contact your local Thermo Fisher representative to learn more about our services and capabilities or request more information.