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A new electrochemical assay method for gene expression using hela cells with a secreted alkaline phosphatase (SEAP) reporter system

Authors

  • Mustafa Şen,

    1. Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan; telephone: +81-22-795-7209; fax: +81-22-795-7209;
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  • Kosuke Ino,

    Corresponding author
    1. Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan; telephone: +81-22-795-7209; fax: +81-22-795-7209;
    • Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan; telephone: +81-22-795-7209; fax: +81-22-795-7209
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  • Hitoshi Shiku,

    1. Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan; telephone: +81-22-795-7209; fax: +81-22-795-7209;
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  • Tomokazu Matsue

    Corresponding author
    1. Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan; telephone: +81-22-795-7209; fax: +81-22-795-7209;
    2. WPI Advanced Institute of Materials Research, Tohoku University, 2-1-1 Katahira, Aoba, Sendai 980-8579, Japan
    • WPI Advanced Institute of Materials Research, Tohoku University, 2-1-1 Katahira, Aoba, Sendai 980-8579, Japan
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Abstract

A new electrochemical assay for the detection of secreted alkaline phosphatase (SEAP) from transfectant HeLa cells is proposed using a microarray device and scanning electrochemical microscopy (SECM). The assay consists of two steps: the first is the incubation of a transfected cell in a microarray culture device covered with a substrate modified with anti-SEAP under physiological conditions without any additives. The array device consists of a 4 × 4 array of microwells having a size of 100 µm × 100 µm (diameter × depth). The second step is SECM measurement of secreted SEAP at the antibody-immobilized substrate. This assay ensures accuracy and intactness because the undesired influence of endogeneous ALP is eliminated and the transfected cells are incubated in a culture device under suitable conditions. We successfully detected the expression of SEAP from intact cells at the single-cell level using this assay. The system is useful as a cell-based gene-expression assay. Biotechnol. Bioeng. 2012; 109:2163–2167. © 2012 Wiley Periodicals, Inc.

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