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- W2523294268 abstract "Therecently proposed bioassay procedure that is based on the substrate inducedquenching (SIQ) of an indicator fluorescence for the measurement of analyteconcentrations is evaluated. In this type of assay a enzynatic reaction and afluorescencequenching interaction are coupled together. Typically, an appropriatedehydrogenase enzyme reduces or oxidises the nicotinamide adenine dinucleotidecofactor. Thechange in the concentration of NADH results in variations in the excitedfluorophore population as observed through fluorescence intensity. This latter aspect isused to monitor substrate (analyte) concentrations.Results on theinvestigation of the substrate induced quenching bioassay method andpossibilities of using it as the basis of (i) a novel enzyme bioassay technique and (ii) anovelbioprobe format are presented. Ethanol was chosen as the model analyte, and anewassay procedure for its measurement was developed.Ageneric theoretical relation is discussed for the observed assay kinetics of substrateinducedquenching (SIQ) and a model is described that includes the effects due todynamic/static quenching of the fluorophore by either the enzyme substrate or product.Thevalidity of the derived model is shown by comparison with experimental results for aSIQ based ethanol assay. The option of running the dehydrogenase reaction so as toconsume NADH rather thangenerate it is also investigated. In order to demonstrate thisapproach acetaldehyde was chosen as the model analyte, and a assay procedure for itsmeasurement wasdeveloped.Thepotential of the SIQ technique for incorporation into biosensor based upon a'reservoir' format was demonstrated through the development of custom opticalinstrumentation and resevoir flowcell. Applicability of the SIQ technique to otherbiosensor formats such as flow-injection analysis and 'dry reagent' technology isdiscussed.The overallapplicability of the SIQ technique is assessed through the generation of anumber of SIQ assays on the following substrates: ethanol, glucose, glucose-6-phosphate, L-glutamic acid, isocitric acid, acetaldehyde, pyruvic acid, ot-ketoglutaricacid, and oxalacetic acid." @default.
- W2523294268 created "2016-09-30" @default.
- W2523294268 creator A5042493129 @default.
- W2523294268 date "1995-08-01" @default.
- W2523294268 modified "2023-09-23" @default.
- W2523294268 title "Optical fluoroassays based on substrate induced quenching" @default.
- W2523294268 hasPublicationYear "1995" @default.
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