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Paper microfluidic‐based enzyme catalyzed double microreactor
Author(s) -
Ferrer Ivonne M.,
Valadez Hector,
Estala Lissette,
Gomez Frank A.
Publication year - 2014
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400091
Subject(s) - resazurin , microreactor , nicotinamide adenine dinucleotide , chemistry , microfluidics , nad+ kinase , catalysis , biocatalysis , chromatography , buffer solution , fluorescence , enzyme catalysis , enzyme , nanotechnology , biochemistry , reaction mechanism , materials science , physics , quantum mechanics
We describe a paper microfluidic‐based enzyme catalyzed double microreactor assay using fluorescent detection. Here, solutions of lactate dehydrogenase (LDH) and diaphorase (DI) were directly spotted onto the microfluidic paper‐based analytical device (μPAD). Samples containing lactic acid, resazurin, and nicotinamide adenine dinucleotide oxidized form (NAD + ), potassium chloride (KCl), and BSA, in MES buffer were separately spotted onto the μPAD and MES buffer flowed through the device. A cascade reaction occurs upon the sample spot overlapping with LDH to form pyruvate and nicotinamide adenine dinucleotide reduced form (NADH). Subsequently, NADH is used in the conversion of resazurin to fluorescent resorufin by DI. The μPAD avoids the need of surface functionalization or enzyme immobilization steps. These microreactor devices are low cost and easy to fabricate and effect reaction based solely on buffer capillary action.