A simple screening assay for cholinesterase activity and inhibition based on optical oxygen detection

Alice Zitova, Fiach C. O'Mahony, Ilya N. Kurochkin, Dmitri B. Papkovsky

Research output: Contribution to journalArticlepeer-review

Abstract

A screening assay for the measurement of activity and inhibition of cholinesterase enzymes is described. It employs a coupled bi-enzymatic reaction of butyrylcholinesterase and choline oxidase and quenched-phosphorescence detection of dissolved oxygen with an oxygen-sensitive probe. The assay can operate in 96-well plates or capillary micro-cuvettes with detection on conventional fluorescent plate reader or the LightCycler® platform, respectively. It was demonstrated with the inhibition assays of paraoxon and carbofuran pesticides for which IC50 values were determined and system performance assessed. This methodology provides moderate sensitivity (nM-μM range of enzyme or inhibitor concentrations), simplicity, sample throughput, and convenience.

Original languageEnglish
Pages (from-to)1746-1755
Number of pages10
JournalAnalytical Letters
Volume43
Issue number10-11
DOIs
Publication statusPublished - 1 Jul 2010

Keywords

  • Acetylcholine esterase inhibitors
  • Carbamates
  • Choline oxidase
  • Enzymatic assays
  • Optical oxygen sensing
  • Organophospates
  • Phosphorescence quenching
  • Screening

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