Gibco Freedom DG44 Kit Frequently Asked Questions

IgG titers of ≥0.5 g/L in simple fed batch and ≥1 g/L in fed batch with complex feeds have been obtained using the Freedom DG44 Kit. Titers of 20-100 mg/L with blood factors and other hard to express proteins (HTEP) have also been achieved with this kit.

It is possible to go from transfection to lead clone in less than 6 months with the Freedom DG44 Kit.

Product quality depends on multiple development aspects, such as the product itself, the selected clone, and the upstream and downstream process. Freedom DG44 cells offer all the typical CHO cell line benefits in respect to low protease levels and preferred glycan pattern.

The stability of expression was assessed for over 60 generations with the observation that most clones maintain their level of expression after 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 DG44 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 to produce products for clinical testing.

All major components to complete the cell line development workflow are included in the Freedom DG44 Kit: cGMP banked DG44 cells, vectors for cloning one or two genes, transfection reagent, competent cells, serum-free cell culture media and reagents. A complete list of components, including additional materials required but not included, can be found in the Freedom DG44 Kit manual.

A certificate of analysis (COA) demonstrating suitability for research use is available to everyone who purchases Freedom DG44 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.

Detailed information about the vectors can be found in the Freedom DG44 Kit manual.

In the DG44 system, Geneticin and DHFR selection are both used, either sequentially or simultaneously.

Freedom DG44 cells have been optimized for use with Gibco CD OptiCHO Medium and the CHO CD EfficientFeed Kit.

Good results can be obtaining using either limiting dilution cloning or ClonePix™ FL. For advice regarding ClonePix™ cloning, see recommendations in the Freedom DG44 Kit manual or request a consultation at pd.direct@lifetech.com

Both FreeStyle MAX Reagent (included in the kit) and the Neon Transfection System provide stable pools with similar final titers.

Only two transfections are recommended

When using limiting dilution cloning, careful evaluation of monoclonality of a clone is required before assessing stability of expression in order to avoid heterogeneous clones that can drift into separate populations and appear unstable.

IgG-expressing DG44 clones readily scale-up from shake flask to 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 DG44 cell line.

To inquire about a commercial license, please email: pd.direct@lifetech.com.

Yes. Contact our field-based Process Science Fellows (PSFs) at pd.direct@lifetech.com for technical support

Yes. The process science staff at Thermo Fisher Scientific can assist you with the development of a customized stable cell line development program, which allows for the rapid screening of high producing clones. To learn more, emailpd.direct@lifetech.com