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

被引：413

作者：

Yamane-Ohnuki, N ^{[1
]}

Kinoshita, S ^{[1
]}

Inoue-Urakubo, M ^{[1
]}

Kusunoki, M ^{[1
]}

Iida, S ^{[1
]}

Nakano, R ^{[1
]}

Wakitani, M ^{[1
]}

Niwa, R ^{[1
]}

Sakurada, M ^{[1
]}

Uchida, K ^{[1
]}

Shitara, K ^{[1
]}

Satoh, M ^{[1
]}

机构：

[1] Kyowa Hakko Kogyo Co Ltd, Tokyo Res Labs, Machida, Tokyo 1948533, Japan

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2004年 / 87卷 / 05期

关键词：

Chinese hamster ovary (CHO) cells; FUT8; knockout; alpha-1,6-fucosylation; therapeutic antibody; antibody-dependent; cellular cytoxicity (ADCC);

D O I：

10.1002/bit.20151

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 alpha-1 6-fucosyltransferase that catalyzes the transfer of fucose from GDP-fucose to N-acetylglucosamine (GlcNAc) in an alpha-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 Fcgamma-receptor IIIa (FcgammaRIIIa) 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 highADCC antibodies with fixed quality and efficacy for therapeutic use. (C) 2004 Wiley Periodicals, Inc.

引用

页码：614 / 622

页数：9

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