Thermoelectric ELISA for quantification of 8OHdG in a microfluidic device

This research demonstrates the feasibility o f a novel method for performing thermoelectric enzyme-linked immunosoibent assay (ELISA) in a microfluidic device. The feasibility o f the thermoelectric ELISA is demonstrated by measuring the concentration o f 8-hydroxy 2-deoxyguanosine (80HdG) in urine samples from amyloid precursor protein (APP) transgenic mice. The detection method is based on formation of a complex between 80HdG and anti-80HdG capture antibody conjugated to biotin. The complex is immobilized over the measuring junctions o f a thermopile via biotin streptavidin interaction. The concentration of the analyte is determined by using enzyme linked secondary IgG antibody specific to the primary one. The concentration of 80HdG is determined by the initiation o f an enzymatic reaction between glucose and glucose oxidase that is conjugated to the secondary IgG antibody. The heat released by the reaction o f glucose and glucose oxidase is measured using an antimony-bismuth thermopile integrated in a microfluidic device. The amount o f heat detected by the sensor is inversely proportional to the concentration o f 80HdG. A standard calibration curve using known concentrations of synthetic 80HdG is generated and used to determine the concentration of the oxidized guanine in mouse urine samples. APPROVAL FOR SCHOLARLY DISSEMINATION The author grants to the Prescott Memorial Library of Louisiana Tech University the right to reproduce, by appropriate methods, upon request, any or all portions of this Dissertation. It is understood that “proper request” consists o f the agreement, on the part of the requesting party, that said reproduction is for his personal use and that subsequent reproduction will not occur without written approval o f the author o f this Dissertation. Further, any portions o f the Dissertation used in books, papers, and other works must be appropriately referenced to this Dissertation. Finally, the author o f this Dissertation reserves the right to publish freely, in the literature, at any time, any or all portions of this Dissertation.