Producing defucosylated antibodies with enhanced in vitro antibody‐dependent cellular cytotoxicity via FUT8 knockout CHO‐S cells |
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| Authors: | Huifang Zong Lei Han Kai Ding Jiaxian Wang Tao Sun Xinyu Zhang Cedric Cagliero Hua Jiang Yueqing Xie Jianrong Xu Baohong Zhang Jianwei Zhu |
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| Institution: | 1. Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People's Republic of China;2. Jecho Laboratories, Frederick, MD, USA;3. School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China |
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| Abstract: | To engineer a host cell line that produces defucosylated mAbs with superior antibody‐dependent cellular cytotoxicity, we disrupted α‐1, 6 fucosyltransferase (FUT8 ) gene in CHO‐S (CHO is Chinese hamster ovary) cells by clustered regularly interspaced short palindromic repeats‐CRISPR associated nuclease 9. The gene knockout cell line was evaluated for growth, stability, and product quality. The growth profile of FUT8 gene knockout CHO‐S (FUT8 ?/?) cells was comparable with wild type CHO‐S cells. FUT8 catalyzes the transfer of a fucose residue from GDP‐fucose to N‐glycans residue. Defucosylated IgG1 antibodies produced by FUT8 ?/? cells showed increased binding affinities to human FcγRIIIa and higher activities in mediating antibody‐dependent cellular cytotoxicity, comparing with conventional fucosylated IgG1. Our results demonstrated the potential of using the clustered regularly interspaced short palindromic repeats‐CRISPR associated nuclease 9 technology in cell line engineering for biopharmaceutical industrial applications. |
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| Keywords: | α ‐1 6 fucosyltransferase (FUT8) Antibody‐dependent cellular cytotoxicity (ADCC) CRISPR/Cas9 Defucosylated antibodies Gene knockout |
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