Genentech has leveraged the ATF6 Luciferase Reporter CHO-K1 Stable Cell Line from Signosis to develop stable cell lines for therapeutic monoclonal antibodies (mAb) production. CHO cells are commonly used for generating stable cell lines to produce therapeutic proteins, and Genentech has used three different CHO host cells for this purpose: dihydrofolate reductase (DHFR) positive CHOK1, DHFR-deficient DG44, and DUXB11-based DHFR deficient CHO.
Genentech's current commercial full-length antibody products have been produced in the DUXB11-derived DHFR-deficient CHO host, but it has been challenging to develop stable cell lines producing sufficient amounts of antibody proteins for some molecules in this host. Therefore, the CHOK1 host has been explored as an alternative approach.
In this study, stable cell lines were developed for three antibody molecules in both DUXB11-based and CHOK1 hosts using the ATF6 Luciferase Reporter CHO-K1 Stable Cell Line from Signosis. The best CHOK1 clones produced approximately 1 g/l for an antibody mAb1 and about 4 g/l for an antibody mAb2 in 14-day fed-batch cultures in shake flasks, while the DUXB11-based host produced only ~0.1 g/l for both antibodies in the same production experiments. For an antibody mAb3, both CHOK1 and DUXB11 host cells generated stable cell lines, with the best clone in each host producing approximately 2.5 g/l.
The ATF6 Luciferase Reporter CHO-K1 Stable Cell Line from Signosis was instrumental in this study, enabling the efficient development and selection of stable cell lines for mAb production in both host cells. The study also revealed that the CHOK1 host cells have a larger endoplasmic reticulum and higher mitochondrial mass, indicating their potential suitability for the production of certain antibody molecules.
Overall, this success story highlights Genentech's expertise in leveraging cutting-edge technologies such as stable cell lines and luciferase reporter systems to overcome challenges in biotech research and development and drive progress towards developing life-changing therapies for patients.
C hinese hamster ovary K1 host cell enables stable cell line development for antibody molecules which are difficult to express in DUXB11-derived dihydrofolate reductase deficient host cell (Article)
Dept. of Early Stage Cell Culture, Genentech Inc., 1 DNA Way, South San Francisco, CA, United States, Novartis Institutes for Biomedical Research, 100 Technology Square, Cambridge, MA 02139, United States
Abstract
Therapeutic monoclonal antibodies (mAb) are often produced in Chinese hamster ovary (CHO) cells. Three commonly used CHO host cells for generating stable cell lines to produce therapeutic proteins are dihydrofolate reductase (DHFR) positive CHOK1, DHFR-deficient DG44, and DUXB11-based DHFR deficient CHO. Current Genentech commercial full-length antibody products have all been produced in the DUXB11-derived DHFR-deficient CHO host. However, it has been challenging to develop stable cell lines producing an appreciable amount of antibody proteins in the DUXB11-derived DHFR-deficient CHO host for some antibody molecules and the CHOK1 host has been explored as an alternative approach. In this work, stable cell lines were developed for three antibody molecules in both DUXB11-based and CHOK1 hosts. Results have shown that the best CHOK1 clones produce about 1 g/l for an antibody mAb1 and about 4 g/l for an antibody mAb2 in 14-day fed batch cultures in shake flasks. In contrast, the DUXB11-based host produced ∼0.1 g/l for both antibodies in the same 14-day fed batch shake flask production experiments. For an antibody mAb3, both CHOK1 and DUXB11 host cells can generate stable cell lines with the best clone in each host producing ∼2.5 g/l. Additionally, studies have shown that the CHOK1 host cell has a larger endoplasmic reticulum and higher mitochondrial mass. © 2013 American Institute of Chemical Engineers.