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

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26 Citations (Scopus)

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 100μm×100μm (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.

Original languageEnglish
Pages (from-to)2163-2167
Number of pages5
JournalBiotechnology and Bioengineering
Volume109
Issue number8
DOIs
Publication statusPublished - 2012 Aug

Keywords

  • Biosensor
  • Cell array
  • Electrochemistry
  • Lab on a chip
  • Reporter gene assay

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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