101. A new electrochemical assay method for gene expression using hela cells with a secreted alkaline phosphatase (SEAP) reporter system
- Author
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Hitoshi Shiku, Kosuke Ino, Tomokazu Matsue, and Mustafa Şen
- Subjects
Reporter gene ,Gene Expression Profiling ,Cell ,Substrate (chemistry) ,Bioengineering ,Electrochemical Techniques ,Transfection ,Biology ,Alkaline Phosphatase ,Microarray Analysis ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Molecular biology ,HeLa ,Scanning electrochemical microscopy ,medicine.anatomical_structure ,Genes, Reporter ,Gene expression ,medicine ,Humans ,Biosensor ,HeLa Cells ,Biotechnology - 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.
- Published
- 2012
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