Methodological Analysis of an Improved Gluten Quantitation Aptamer-based Biosensor
Date
2022-09-22
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Abstract
Fluorescence resonance energy transfer based aptamer are currently being studied in many
research groups due to their potential contribution as an alternative gluten standard for gluten-
quantitation. The current gold standard withholds critical limitations due to the nature of the
system, hence this aptamer-based sensor (or aptasensor for short) application represents one
of the alternatives; yet, drawbacks such as low signal-to-noise ratio and reliability are in the
scope of research groups aiming to overcome them. In this study, multiple variations of the protocol are assessed based on correctly classifying
food samples as their actual concentration of gluten, this is coined as the accuracy of the
biosensor. The study also aims to overcome the current limitation of the gold standard in
fermented samples by including soy sauce and malt vinegar in the tests. And different additives
aiming to help overcoming the limitation were implemented into the protocol and assessed. This approach allowed the biosensor to classify the products with 98.28% of accuracy,
and 0% of error in classifying gluten-rich products (false-negatives) within the first 3 days
of bioassay preparation; yet, this bioassay needs to be studied further as only 18 different
off-the-shelf products were tested (over 800 tests in total). Additionally, after the first week,
false-negatives increased to around 5% and remained that way until the end of the first month. The cause of this relies mostly on the decomposition of the conjugate reduced graphene
oxide and polyethylene glycol that are implemented in the system. This implies that further
research aiming solely at additives or alternative reagents that increase the lifespan or stability
of the conjugate would augment the overall performance.
Description
Keywords
Aptamer, Fluorescence, FRET, Gluten sensor, 33-mer peptide, Aptasensor, Graphene oxide, ELISA
Citation
Kuri Martinez, J. C. (2022). Methodological analysis of an improved gluten quantitation aptamer-based biosensor (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.