title: Novel electrochemical methodology for activity estimation of alkaline phosphatase based on solubility difference.
abstract: We propose a novel electrochemical detection system for alkaline phosphatase (ALP) activity using the difference in water and oil solubilities between the substrate, ferrocene ethyl phosphate ester (FcEtOPO(3)(2-)), and the enzymatic product, ferroceneethanol (FcEtOH). In this system, water droplets containing ALP and FcEtOPO(3)(2-) were placed on a Pt disk microelectrode and surrounded by a mineral oil. By the ALP-catalyzed reaction, FcEtOPO(3)(2-) was converted to FcEtOH, which was then transferred to the mineral oil from the water droplets with FcEtOPO(3)(2-) remaining in the water droplets. After partitioning FcEtOH from the water droplets, FcEtOPO(3)(2-) was detected at the Pt disk microelectrode to estimate the ALP activity. Using this novel system, the ALP activity of embryoid bodies was successfully detected. We believe that the present system will be widely applicable to ALP-based bioassays.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, Sendai, Japan. ino.kosuke@bioinfo.che.tohoku.ac.jp
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ino
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kanno
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Arai
authors.forename: Kumi Y
authors.initials: KY
authors.lastname: Inoue
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis* / Q000378:metabolism*; D000818:Animals; D055664:Electrochemical Techniques; D004566:Electrodes; D053595:Embryonic Stem Cells / Q000201:enzymology; D004952:Esters; D000431:Ethanol / Q000737:chemistry; D005296:Ferrous Compounds / Q000737:chemistry; D000075163:Metallocenes; D051379:Mice; D008899:Mineral Oil / Q000737:chemistry; D010984:Platinum / Q000737:chemistry; D012995:Solubility; D014867:Water / Q000737:chemistry
chemical_list: D004952:Esters; D005296:Ferrous Compounds; D000075163:Metallocenes; D014867:Water; D000431:Ethanol; D010984:Platinum; D008899:Mineral Oil; D000469:Alkaline Phosphatase; C004998:ferrocene