Simplified Gluten Detection Approach in the Presence of a Black Hole Quencher (BHQ)
On demand starting 10 October 2021
An efficient method to bind Black Hole Quencher (BHQ) to gliadin from gluten has been developed. In the present work, we used the physical phenomenon of fluorescence resonance energy transfer (FRET) to study the quenching and hybridization efficiency of BHQ to gliadin. For this process, we developed an experiment with a fluorophore (6-Carboxyfluorescein or 6-FAM) labeled gliadin specific aptamer. The gliadin from gluten was labeled with BHQ using two different reactions: BHQ-10 succinimidyl ester (NHS ester) and BHQ-10 carboxylic acid. The reaction of BHQ-10 carboxylic acid with gliadin demonstrated to be efficient for quenching the fluorophore energy by an average of 62% in comparison to the gliadin as a control. Additionally, we demonstrated the hybridization efficiency by the FRET physical phenomenon. This sets the basis for the breakthrough development of a simplified gluten detection biosensor by the elimination of the fluorophore in common biosensing systems.
Samaria Nevarez Diaz
Univ. of Calgary (Canada)
Ms. Samaria Nevarez Diaz, UofC (MSc Student, Lead in Optical Design and assay experiments) previously worked in a system integration company for four years after his B.Sc. She has worked on optical sol-gel based absorbance-type optical filters for fluorescence measurements and is involved in aptamer-assay development and sample extraction optimization. Samaria will lead the project in terms of system design and optical sensing, and the consolidation of multicomponent systems together with other students.