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

doi:10.1080/00032711003653833

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

School of Biochemistry and Cell Biology

Research output: Contribution to journal › Article › peer-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 language	English
Pages (from-to)	1746-1755
Number of pages	10
Journal	Analytical Letters
Volume	43
Issue number	10-11
DOIs	https://doi.org/10.1080/00032711003653833
Publication status	Published - Jul 2010

Keywords

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

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10.1080/00032711003653833

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title = "A simple screening assay for cholinesterase activity and inhibition based on optical oxygen detection",

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{\textregistered} 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.",

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T1 - A simple screening assay for cholinesterase activity and inhibition based on optical oxygen detection

AU - Zitova, Alice

AU - O'Mahony, Fiach C.

AU - Kurochkin, Ilya N.

AU - Papkovsky, Dmitri B.

PY - 2010/7

Y1 - 2010/7

N2 - 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.

AB - 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.

KW - Acetylcholine esterase inhibitors

KW - Carbamates

KW - Choline oxidase

KW - Enzymatic assays

KW - Optical oxygen sensing

KW - Organophospates

KW - Phosphorescence quenching

KW - Screening

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DO - 10.1080/00032711003653833

M3 - Article

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SN - 0003-2719

VL - 43

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EP - 1755

JO - Analytical Letters

JF - Analytical Letters

IS - 10-11

ER -

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

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Keywords

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