Cell-based electrochemical assay for endotoxin using a secreted alkaline phosphatase reporter system

Kumi Y. Inoue; Tomoyuki Yasukawa; Hitoshi Shiku; Tomokazu Matsue

doi:10.5796/electrochemistry.76.525

Cell-based electrochemical assay for endotoxin using a secreted alkaline phosphatase reporter system

Kumi Y. Inoue, Tomoyuki Yasukawa, Hitoshi Shiku, Tomokazu Matsue

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

We developed a cell-based assay device for the detection of endotoxin, the potentially toxic compound that induces septic shock. Genetically-engineered cells that secrete alkaline phosphatase (SEAP) on exposure to endotoxin were cultured in an electrochemical cell device in medium containing p-aminophenyl phosphate and various concentrations of endotoxin. After 24 hr incubation, p-aminophenol (pAP), generated by SEAP-catalyzed hydrolysis, was detected by amperometry at + 0.35 V. The amperometric response increased with the concentration of endotoxin in the range of 0.01-1 ng/ml.

Original language	English
Pages (from-to)	525-528
Number of pages	4
Journal	Electrochemistry
Volume	76
Issue number	8
DOIs	https://doi.org/10.5796/electrochemistry.76.525
Publication status	Published - 2008 Aug

Keywords

Amperometry
Cell-based sensor
Lipopolysaccharide
Secreted alkaline phosphatase

ASJC Scopus subject areas

Electrochemistry

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10.5796/electrochemistry.76.525

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abstract = "We developed a cell-based assay device for the detection of endotoxin, the potentially toxic compound that induces septic shock. Genetically-engineered cells that secrete alkaline phosphatase (SEAP) on exposure to endotoxin were cultured in an electrochemical cell device in medium containing p-aminophenyl phosphate and various concentrations of endotoxin. After 24 hr incubation, p-aminophenol (pAP), generated by SEAP-catalyzed hydrolysis, was detected by amperometry at + 0.35 V. The amperometric response increased with the concentration of endotoxin in the range of 0.01-1 ng/ml.",

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AU - Inoue, Kumi Y.

AU - Yasukawa, Tomoyuki

AU - Shiku, Hitoshi

AU - Matsue, Tomokazu

PY - 2008/8

Y1 - 2008/8

N2 - We developed a cell-based assay device for the detection of endotoxin, the potentially toxic compound that induces septic shock. Genetically-engineered cells that secrete alkaline phosphatase (SEAP) on exposure to endotoxin were cultured in an electrochemical cell device in medium containing p-aminophenyl phosphate and various concentrations of endotoxin. After 24 hr incubation, p-aminophenol (pAP), generated by SEAP-catalyzed hydrolysis, was detected by amperometry at + 0.35 V. The amperometric response increased with the concentration of endotoxin in the range of 0.01-1 ng/ml.

AB - We developed a cell-based assay device for the detection of endotoxin, the potentially toxic compound that induces septic shock. Genetically-engineered cells that secrete alkaline phosphatase (SEAP) on exposure to endotoxin were cultured in an electrochemical cell device in medium containing p-aminophenyl phosphate and various concentrations of endotoxin. After 24 hr incubation, p-aminophenol (pAP), generated by SEAP-catalyzed hydrolysis, was detected by amperometry at + 0.35 V. The amperometric response increased with the concentration of endotoxin in the range of 0.01-1 ng/ml.

KW - Amperometry

KW - Cell-based sensor

KW - Lipopolysaccharide

KW - Secreted alkaline phosphatase

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Cell-based electrochemical assay for endotoxin using a secreted alkaline phosphatase reporter system

Abstract

Keywords

ASJC Scopus subject areas

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