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Capacitive biosensor for detection of endotoxin
Authors:Warakorn Limbut  Martin Hedström  Panote Thavarungkul  Proespichaya Kanatharana  Bo Mattiasson
Affiliation:(1) Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Box 124, 221 00, Lund, Sweden;(2) Biophysics Research Unit: Biosensors and Biocurrents, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand;(3) Department of Chemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand;(4) Department of Physics, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand
Abstract:A capacitive biosensor for the detection of bacterial endotoxin has been developed. Endotoxin-neutralizing protein derived from American horseshoe crab was immobilized to a self-assembled thiol layer on a biosensor transducer (Au). Upon injection of a sample containing endotoxin, a decrease in the observed capacitive signal was registered. Endotoxin could be determined under optimum conditions with a detection limit of 1.0 × 10−13 M and linearity ranging from 1.0 × 10−13 to 1.0 × 10−10 M. Good agreement was achieved when applying endotoxin preparations purified from an Escherichia coli cultivation to the capacitive biosensor system, utilizing the conventional method for quantitative endotoxin determination, the Limulus amebocyte lysate test as a reference. The capacitive biosensor method was statistically tested with the Wilcoxon signed rank test, which proved the system is acceptable for the quantitative analysis of bacterial endotoxin (P < 0.05). MediaObjects/216_2007_1443_Figa_HTML.gif Figure The flow-injection capacitive biosensor system and the capacitive properties of the transducer surface, where CSAM is the capacitance change of the self-assembled thiol monolayer, CP is the capacitance change of the protein layer, Ca is the capacitance change of the analyte layer and CTotal is the total capacitance change measured at the working electrode/solution interface (modified from Limbut et al., 2006. Biosens Bioelectron 22: 233-240)
Keywords:Capacitive biosensor  Endotoxin   Limulus   Self-assembled monolayer  Monolayer
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