Development of an amperometric biosensing method for the determination of L-fucose in pretreated urine

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dc.contributor.authorTsiafoulis, C. G.en
dc.contributor.authorProdromidis, M. I.en
dc.contributor.authorKarayannis, M. I.en
dc.date.accessioned2015-11-24T16:56:02Z
dc.date.available2015-11-24T16:56:02Z
dc.identifier.issn0956-5663-
dc.identifier.urihttps://olympias.lib.uoi.gr/jspui/handle/123456789/10382
dc.rightsDefault Licence-
dc.subjectfucose biosensing methoden
dc.subjectpretreated urineen
dc.subjectpolyvinyl alcohol-styrylpyridiniumen
dc.subjectfucose dehydrogenaseen
dc.subjectlead dioxideen
dc.subjectferricyanideen
dc.subjectperformance liquid-chromatographyen
dc.subjectalpha-l-fucoseen
dc.subjectaciden
dc.subjectglucoseen
dc.subjectserumen
dc.subjectassayen
dc.subjectelectrophoresisen
dc.subjectglycoproteinsen
dc.subjectderivativesen
dc.subjectpyruvateen
dc.titleDevelopment of an amperometric biosensing method for the determination of L-fucose in pretreated urineen
heal.abstractThe first amperometric biosensing method for the determination of L-fucose is described. L-Fucose is the objective of much current research, as it is considered as a potential marker for various pathologic disorders. Recombinant L-fucose dehydrogenase, having as cofactor beta-nicotinamide adenine dinucleotide phosphate (NAD(+)P), was cross-linked in a water-soluble photosensitive polymer matrix, that is, polyvinyl alcohol (PVA) modified with styrylpyridinium (SbQ), in the presence of BSA and glutaraldehyde. The resulting membrane was sandwiched between two polycarbonate membranes and was mounted in an amperometric cell. The oxidation of the enzymatically produced NADPH was monitored at a platinum anode at +0.25 V versus a silver pseudoreference electrode in the presence of ferricyanide. The system was fully optimized with respect to various analytical parameters. Regarding to the mechanical properties of the membrane and the storage stability of the immobilized enzyme, various parameters were also optimized. Several methods for the pretreatment of urine samples were investigated. Treatment of the samples with PbO2 found to eliminate the interference effect of various electroactive species exist in urine; optimum incubation time was determined since at prolonged incubation times L-fucose is also affected. Calibration curves for the direct and the mediated monitoring of NADPH were liner over the concentration ranges 0.04-1.0 mM (r(2) = 0.9995) and 0.03-3.0 mM (r(2) = 0.9997) fucose, respectively. The detection limits (S/N 3) were 2 and 1.5 muM fucose, respectively. The R.S.D. of the mediated biosensor is better than 1.5% (n = 10, 0.5 mM fucose). The proposed biosensor correlates well with a reference enzymatic method and exhibits very good working and storage stability. (C) 2004 Elsevier B.V. All rights reserved.en
heal.accesscampus-
heal.fullTextAvailabilityTRUE-
heal.identifier.primaryDOI 10.1016/j.bios.2004.03.012-
heal.identifier.secondary<Go to ISI>://000225009000030-
heal.identifier.secondaryhttp://ac.els-cdn.com/S0956566304001393/1-s2.0-S0956566304001393-main.pdf?_tid=a56a93e3f78dcd234fb4748014f993fe&acdnat=1333037431_0cc005ff4c31b387cfff58686017cbd9-
heal.journalNameBiosens Bioelectronen
heal.journalTypepeer reviewed-
heal.languageen-
heal.publicationDate2004-
heal.publisherElsevieren
heal.recordProviderΠανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείαςel
heal.typejournalArticle-
heal.type.elΆρθρο Περιοδικούel
heal.type.enJournal articleen

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