Cooverexpression of alanine aminotransferase 1 in Chinese hamster ovary cells overexpressing taurine transporter further stimulates metabolism and enhances product yield

Authors

  • Hisahiro Tabuchi,

    Corresponding author
    1. Chugai Pharmaceutical Co., Ltd., API Process Development Department (Bio Technology), 5-5-1, Ukima, Kita-ku, Tokyo 115-8543, Japan; telephone: 81-3-3968-8602; fax: 81-3-3968-2038
    • Chugai Pharmaceutical Co., Ltd., API Process Development Department (Bio Technology), 5-5-1, Ukima, Kita-ku, Tokyo 115-8543, Japan; telephone: 81-3-3968-8602; fax: 81-3-3968-2038.
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  • Tomoya Sugiyama

    1. Chugai Pharmaceutical Co., Ltd., API Process Development Department (Bio Technology), 5-5-1, Ukima, Kita-ku, Tokyo 115-8543, Japan; telephone: 81-3-3968-8602; fax: 81-3-3968-2038
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  • Hisahiro Tabuchi and Tomoya Sugiyama contributed equally to this work.

Abstract

Innovation in monoclonal antibody (mAb) production continues to be driven by cell engineering strategies to increase yield and improve product quality. In a previous study, to investigate the effectiveness of transporter overexpression strategies, we prepared a taurine transporter-overexpressing host cell line (DXB11/TAUT) that produced a higher proportion of high-mAb-titer strains than did the parent host cell line. In the current study, we selected a single DXB11/TAUT/mAb1 strain that remained viable for longer (up to 1 month) under common fed-batch culture conditions, and the improvement in viability could be attributed to its improved metabolic properties. It was also more productive (up to >100 pg/cell/day) and yielded more mAb1 (up to 8.1 g/L/31 days) than the parent cell line, and the mAb1 it produced was of comparable quality. These results suggested that this host cell engineering strategy has unique potential for the improvement of mAb-producing Chinese hamster ovary (CHO) cells; for example, it may be appropriate for high cell density perfusion culture. TAUT-overexpressing cell lines rapidly accumulated the byproduct alanine, and our challenge in the present study was to apply a strategy for modulating cell metabolism to utilize this byproduct to achieve a high mAb yield in a shorter culture period. To accomplish this, we genetically modified the DXB11/TAUT/mAb1 strain to cooverexpress alanine aminotransferase 1 (ALT1). The resulting DXB11/TAUT/mAb1/ALT1 cooverexpressing strain gave a higher mAb yield in a shorter culture period (5.9 g/L/14 days). It is usually difficult to drive the overexpression of two functional genes while balancing competing goals. However, forced cooverexpression of TAUT and ALT1 in our DXB11/TAUT/mAb1/ALT1 strain resulted in a higher proliferation than the DXB11/TAUT/mAb1 strain, with an ideal balance between cell viability and productivity. Therefore, we have demonstrated a strategy capable of achieving an optimum balance among the goals of cell viability, productivity, and proliferative capacity. Biotechnol. Bioeng. 2013; 110: 2208–2215. © 2013 Wiley Periodicals, Inc.

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