title: A new electrochemical assay method for gene expression using HeLa cells with a secreted alkaline phosphatase (SEAP) reporter system.
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.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki, Aoba, Sendai 980-8579, Japan.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Şen
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ino
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D055664:Electrochemical Techniques / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D046228:Microarray Analysis; D014162:Transfection
chemical_list: D000469:Alkaline Phosphatase