Yadid-Pecht, OrlyTurner, Raymond JosephVastarey, Nikhil Suresh2019-04-082019-04-082019-04-05Vastarey, N. S. (2019). Fluorescence-based gluten detection using aptamer hybrid (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.http://hdl.handle.net/1880/110145This thesis describes different facets of the Gluten detection project. This interdisciplinary project contains aptamer bio-chemistry, sol-gel based optical filters and has explored immobilization techniques for gluten sensing. The aptamer biochemistry contains a single stranded oligonucleotide in combination with PEG and Graphene oxide to enable fluorogenic sensing of gluten. Different food matrices are explored where changes in gluten detection ability can be seen with different aptamer hybrids. The protocol for optimizing the aptamer assay, establishing standard gliadin standards and extraction procedures were optimized. On optimizing the assay parameters, the aptamer sensor is then evolved through different strategies, and its effects are seen on food sample testing. To move towards a strategy for room temperature storage and microfluidic-integration, different immobilization strategies are explored. These include sol-gel based aptamer encapsulation, and aptamer immobilization on an amine-based sol-gel hybrid. The efficiency of binding is gauged through fluorescent experiments on epi-fluorescent microscope. To miniaturize the instrumentation related to fluorescence detection, this thesis implements droplet-based absorbance filters using sol-gel polymerization technique. In accordance with the Beer-Lambert’s law, different dyes are used to obtain absorbance spectra to enable fluorescence transmission into the CMOS imager. A dye is eventually selected, and aptamer-gliadin binding experiments illustrate the miniaturized optical system’s ability to differentiate gluten’s concentrationengUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.glutenaptamerfluorescence tagged aptamergliadinsol-gel optical filterabsorbance filterEngineeringmaster thesis10.11575/PRISM/36343