Journal
INNATE IMMUNITY
Volume 18, Issue 2, Pages 343-349Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1177/1753425911410337
Keywords
Biosensor; carbon electrode; endotoxin assay; lipopolysaccharide; voltammetry
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Funding
- Japan Society for the Promotion of Science (JSPS) [22245001]
- Japan Science and Technology Agency
- Grants-in-Aid for Scientific Research [23750073, 22245011] Funding Source: KAKEN
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Here, we report the development of an electrochemical detection method for endotoxin based on the Limulus amebocyte lysate (LAL) assay. A mixture of LAL reagent and endotoxin sample solution was incubated for 1 h. The endotoxin activated a cascade reaction of zymogens contained in the LAL to generate p-nitroaniline (pNA) which was then electrochemically detected by differential pulse voltammetry (DPV). The generated pNA gave a clear peak at -0.75 V vs. silver/silver chloride (Ag/AgCl), which increased with the concentration of endotoxin in the LAL assay solution. This DPV detection was performed using an electrode chip device fabricated from a diamond-like carbon-coated glass substrate. This chip device could detect as low as 10 endotoxin units l(-1) at room temperature within 1 h. This novel electrochemical method for the detection of endotoxin appears promising for the development of compact, low-cost and easy-to-use sensors for on-site monitoring of potentially contaminated medical supplies, including dialysis fluid, transplanted tissue and culture medium for assisted reproduction.
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