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

作者：Sen Mustafa; Ino Kosuke^*; Shiku Hitoshi; Matsue Tomokazu

来源：Biotechnology and Bioengineering, 2012, 109(8): 2163-2167.

DOI：10.1002/bit.24461

• electrochemistry
• biosensor
• lab on a chip
• reporter gene assay
• cell array

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?x?4 array of microwells having a size of 100?mu m?x?100?mu m (diameter?x?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; 109:21632167.

• 出版日期2012-8