Establishment of FUT8 knockout Chinese hamster ovary cells: An ideal host cell line for producing completely defucosylated antibodies with enhanced antibody-dependent cellular cytotoxicity

Authors

  • Naoko Yamane-Ohnuki,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Satoko Kinoshita,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Miho Inoue-Urakubo,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Machi Kusunoki,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Shigeru Iida,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Ryosuke Nakano,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Masako Wakitani,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Rinpei Niwa,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Mikiko Sakurada,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Kazuhisa Uchida,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Kenya Shitara,

    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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  • Mitsuo Satoh

    Corresponding author
    1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
    • Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan; telephone: 81-42-725-2556; fax: 81-42-725-8330
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Abstract

To generate industrially applicable new host cell lines for antibody production with optimizing antibody-dependent cellular cytotoxicity (ADCC) we disrupted both FUT8 alleles in a Chinese hamster ovary (CHO)/DG44 cell line by sequential homologous recombination. FUT8 encodes an α-1,6-fucosyltransferase that catalyzes the transfer of fucose from GDP-fucose to N-acetylglucosamine (GlcNAc) in an α-1,6 linkage. FUT8−/− cell lines have morphology and growth kinetics similar to those of the parent, and produce completely defucosylated recombinant antibodies. FUT8−/−-produced chimeric anti-CD20 IgG1 shows the same level of antigen-binding activity and complement-dependent cytotoxicity (CDC) as the FUT8+/+-produced, comparable antibody, Rituxan. In contrast, FUT8−/−-produced anti-CD20 IgG1 strongly binds to human Fcγ-receptor IIIa (FcγRIIIa) and dramatically enhances ADCC to approximately 100-fold that of Rituxan. Our results demonstrate that FUT8−/− cells are ideal host cell lines to stably produce completely defucosylated high-ADCC antibodies with fixed quality and efficacy for therapeutic use. © 2004 Wiley Periodicals, Inc.

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