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Fabrication of a Dual-Enzyme Sensing System Based on Surface Plasmon Resonance Imager (SPRi) for Simultaneous Determination of Bio-Markers

Yuhei Hida and Hiroaki Shinohara

Saccharides such as glucose, galactose and amino acids like lysine, phenylalanine are known as bio-markers of metabolic disorders, and several analytical methods have been developed to determine these bio-markers in blood and urine by enzymatic method, HPLC analysis and so on. Multi-biosensing systems for simultaneous determination of these bio-markers are presently expected to be necessary for troublesome diagnosis. We have previously proposed enzyme-sensors based on a two-dimensional surface plasmon resonance imager (2D-SPRi), which can sensitively monitor refractive index change of local region in the evanescent field on an Au chip. L-lysine sensor, L-glutamate sensor and galactose sensor has been developed each by combining oxidase reactions with 2D-SPR imaging of Os polymer-redox state. In this study, we aimed to develop a dual-enzyme sensor capable of the simultaneous detection of lysine and galactose by means of co-immobilizing LysOx/Os-HRP polymer spot and GalOx/Os-HRP polymer spot adjacently on the Au chip, which is divided into two sections.