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Comparison of cell lines for stable production of fucose‐negative antibodies with enhanced ADCC

2006; Wiley; Volume: 94; Issue: 4 Linguagem: Inglês

10.1002/bit.20880

1097-0290

Yutaka Kanda, Naoko Yamane‐Ohnuki, Naoto Sakai, Kazuya Yamano, Ryosuke Nakano, Miho Inoue, Hirofumi Misaka, Shigeru Iida, Masako Wakitani, Yoshinobu Konno, Keiichi Yano, Kenya Shitara, Shinji Hosoi, Mitsuo Satoh,

Transgenic Plants and Applications

Abstract Several methods have been described to enhance antibody‐dependent cellular cytotoxicity (ADCC) using different host cells that produce antibody with reduced levels of fucose on their carbohydrates. We compared the suitability of these methods for the serum‐free fed‐batch production of antibody for clinical trials and commercial uses. Recombinant anti‐human CD20 chimeric IgG1‐producing clones were established from host‐cells that have been shown to produce more than 90% fucose‐negative antibody. The cell lines were a FUT8 (α‐1,6‐fucosyltransferase) knockout Chinese hamster ovary (CHO) cell line, Ms704, and two Lens culinaris agglutinin (LCA)‐resistant cell lines, one derived from a variant CHO line, Lec13 and the other from a rat hybridoma cell line, YB2/0. The amount of fucose‐negative antibody produced by Lec13 and YB2/0 significantly decreased with the culture. The increase in fucosylation was due to remaining synthesis of GDP‐fucose via de novo pathway for the CHO line and the elevation of FUT8 expression by the YB2/0 cells. In contrast, Ms704 cells stably produced fucose‐negative antibody with a consistent carbohydrate structure until the end of the culture. The productivity of the Ms704 cells reached 1.76 g/L with a specific production rate (SPR) of 29 pg/cell/day for 17 days in serum‐free fed‐batch culture using a 1 L spinner bioreactor. Our results demonstrate that FUT8 knockout has the essential characteristics of host cells for robust manufacture of fucose‐negative therapeutic antibodies with enhanced ADCC. © 2006 Wiley Periodicals, Inc.