Features of the Freedom® DG44 Kit


Q: What titers can be achieved with the Freedom® DG44 Kit?


A: 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.


Q: How long does it take to obtain a lead clone?


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


Q: What level of product quality is seen with Freedom® DG44 Kit?


A: 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.


Q: How successful is the Freedom® DG44 Kit in creating stable clones?


A: 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.


Q: Can DG44 cells be used for the production of commercial products?


A: 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.



Q: What is included in the Freedom® DG44 Kit?


A: 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.


Q: Does the kit contain documentation of the DG44 cell line?


A: 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.


Q: Where are the Freedom® DG44 Kit vector maps located?


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


Q: What selection system is used in the Freedom® DG44 Kit?


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

Recommended Methods


Q: What cell culture medium and feed do you recommend for Freedom® DG44 cells?


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


Q: What cloning methods can be used with the Freedom® DG44 Kit?


A: 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.


Q: What transfection methods can be used with the Freedom® DG44 Kit?


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


Q: How many transfections are typically required to get high-expressing clones from the Freedom® DG44 Kit?


A: Only two transfections are recommended.


Q: What are some considerations for obtaining a high number of stable clones?


A: 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.


Q: What are some considerations for successful scale-up of DG44 cells?


A: IgG-expressing DG44 clones readily scale-up from shake flask to bioreactor vessels while maintaining growth and titer performance.

Licensing of the DG44 Cell Line


Q: When is a license required for the Freedom® DG44 Kit?


A: 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.


Q: Is there a pull-through royalty on final products or an annual maintenance fee?


A: 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.


Q: How can I learn more about commercial licensing of Freedom® DG44 cells?


A: To inquire about a commercial license, please email: pd.direct@lifetech.comTo read the complete list of FAQs related to licensing, click here.

Technical support and cell line development services


Q: Do you offer technical support for the Freedom® DG44 Kit?


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


Q: Do you offer cell line development services?


A: Yes. Life Technologies’ process science staff 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, email pd.direct@lifetech.com.