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Generation of FX-/- and Gmds-/- CHOZN host cell lines for the production of afucosylated therapeutic antibodies.

Biotechnology progress (2020-08-05)
Weiyi Liu, Roshan Padmashali, Omar Quintero Monzon, Dianna Lundberg, Shan Jin, Brian Dwyer, Yun-Jung Lee, Anisha Korde, Sophia Park, Clark Pan, Bohong Zhang
ABSTRACT

Antibody-dependent cellular cytotoxicity (ADCC) is the primary mechanism of actions for several marketed therapeutic antibodies (mAbs) and for many more in clinical trials. The ADCC efficacy is highly dependent on the ability of therapeutic mAbs to recruit effector cells such as natural killer cells, which induce the apoptosis of targeted cells. The recruitment of effector cells by mAbs is negatively affected by fucose modification of N-Glycans on the Fc; thus, utilization of afucosylated mAbs has been a trend for enhanced ADCC therapeutics. Most of afucosylated mAbs in clinical or commercial manufacturing were produced from Fut8-/- Chinese hamster ovary cells (CHO) host cells, generally generating low yields compared to wildtype CHO host. This study details the generation and characterization of two engineered CHOZN® cell lines, in which the enzyme involved in guanosine diphosphate (GDP)-fucose synthesis, GDP mannose-4,6-dehydratase (Gmds) and GDP-L-fucose synthase (FX), was knocked out. The top host cell lines for each of the knockouts, FX-/- and Gmds-/-, were selected based on growth robustness, bulk MSX selection tolerance, production titer, fucosylation level, and cell stability. We tested the production of two proprietary IgG1 mAbs in the engineered host cells, and found that the titers were comparable to CHOZN® cells. The mAbs generated from either KO cell line exhibited loss of fucose modification, leading to significantly boosted FcγRIIIa binding and ADCC effects. Our data demonstrated that both FX-/- and Gmds-/- host cells could replace Fut8-/- CHO cells for clinical manufacturing of antibody therapeutics.

MATERIALS
Product Number
Brand
Product Description

SAFC
EX-CELL® Advanced CHO Fed-batch Medium, liquid
SAFC
EX-CELL® CD CHO Fusion, Animal-component free, without L-glutamine, without hypoxanthine, without thymidine, liquid, sterile-filtered, suitable for cell culture
SAFC
CHOZN® CHO K1 Host Cell Line, Suspension-adapted in CD media