Miniaturized amperometric biosensor based on xanthine oxidase for monitoring hypoxanthine in cell culture media

Analytical Biochemistry
L MaoL Jin

Abstract

Fabrication and characterization of miniaturized amperometric hypoxanthine biosensors are described and demonstrated for monitoring hypoxanthine in myocardial cell culture media. The sensors are based on xanthine oxidase (XO) immobilized on carbon fiber microelectrodes (CFMEs) using a composite film of Nafion and electropolymerized phenol (PPh). Nafion was used for XO immobilization because of its film hydrophobicity, enzyme-favored environment, and electrostatic interaction with XO, which was dispersed in Nafion film by immersing the Nafion-coated CFMEs in XO solution for 5 h. PPh film was formed as an overlay on Nafion and XO-modified CFMEs via electropolymerization. Hypoxanthine was measured with the sensor by the oxidation of enzymatic reaction products, hydrogen peroxide (H(2)O(2)), and uric acid (UA) at +0.60 V (vs Ag/AgCl). The use of Nafion and PPh as a matrix for XO immobilization yields enhanced specificity, sensitivity, and linearity toward hypoxanthine. A dynamic linear range of 5.0 microM to 1.8 mM was achieved with a calculated detection limit of 1.5 microM (S/N = 3) and a sensitivity of 3.144 nA/mM. In addition, the measurement was virtually interference-free from easily oxidizable species such as UA, ascorbic ac...Continue Reading

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Citations

Mar 19, 2014·Enzyme and Microbial Technology·Chandra Shekhar Pundir, Rooma Devi
Dec 3, 2014·Analytica Chimica Acta·Tânia S C R RebeloM H Fernandes
Oct 6, 2005·Biosensors & Bioelectronics·Yang TianTakeo Ohsaka
Feb 24, 2009·Biochemistry. Biokhimii︠a︡·M R RashidiA Jouyban
Dec 25, 2012·Analytical and Bioanalytical Chemistry·A Carolina TorresChristopher M A Brett
May 31, 2017·Enzyme and Microbial Technology·Ali HalalipourJosé I Reyes-De-Corcuera
Mar 3, 2011·The Journal of Physical Chemistry. B·Palraj KalimuthuPaul V Bernhardt
Sep 1, 2012·The Journal of Physical Chemistry. B·Palraj KalimuthuPaul V Bernhardt
Nov 9, 2012·Analytical Chemistry·Palraj KalimuthuPaul V Bernhardt

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