title: Chinese 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.
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.
authors.affiliation: Dept. of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA 94080, USA.
authors.forename: Zhilan
authors.initials: Z
authors.lastname: Hu
authors.forename: Donglin
authors.initials: D
authors.lastname: Guo
authors.forename: Shirley S M
authors.initials: SS
authors.lastname: Yip
authors.forename: Dejin
authors.initials: D
authors.lastname: Zhan
authors.forename: Shahram
authors.initials: S
authors.lastname: Misaghi
authors.forename: John C
authors.initials: JC
authors.lastname: Joly
authors.forename: Bradley R
authors.initials: BR
authors.lastname: Snedecor
authors.forename: Amy Y
authors.initials: AY
authors.lastname: Shen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016466:CHO Cells; D002478:Cells, Cultured; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005434:Flow Cytometry; D008928:Mitochondria / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D013762:Tetrahydrofolate Dehydrogenase
keywords: CHO cells; CHOK1; DUXB11; antibody