Fluorescence-based gluten detection using aptamer hybrid
Date
2019-04-05
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Abstract
This 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 concentration
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Keywords
gluten, aptamer, fluorescence tagged aptamer, gliadin, sol-gel optical filter, absorbance filter
Citation
Vastarey, N. S. (2019). Fluorescence-based gluten detection using aptamer hybrid (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.