Novel electrochemical methodology for activity estimation of alkaline phosphatase based on solubility difference

Kosuke Ino, Yusuke Kanno, Toshiharu Arai, Kumi Y. Inoue, Yasufumi Takahashi, Hitoshi Shiku, Tomokazu Matsue

Research output: Contribution to journalArticlepeer-review

55 Citations (Scopus)

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 (FcEtOPO3 2-), and the enzymatic product, ferroceneethanol (FcEtOH). In this system, water droplets containing ALP and FcEtOPO32- were placed on a Pt disk microelectrode and surrounded by a mineral oil. By the ALP-catalyzed reaction, FcEtOPO32- was converted to FcEtOH, which was then transferred to the mineral oil from the water droplets with FcEtOPO32- remaining in the water droplets. After partitioning FcEtOH from the water droplets, FcEtOPO32- 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.

Original languageEnglish
Pages (from-to)7593-7598
Number of pages6
JournalAnalytical Chemistry
Volume84
Issue number18
DOIs
Publication statusPublished - 2012 Sep 18

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint Dive into the research topics of 'Novel electrochemical methodology for activity estimation of alkaline phosphatase based on solubility difference'. Together they form a unique fingerprint.

Cite this