Poster number 78 submitted by Volition La
Fluorescence Enhancing Aptamers in Biosensing Applications
Volition La (University of Waterloo, Canada), Thorsten Dieckmann (University of Waterloo, Canada)
Presently, an aptamer that binds to a derivative of thiazole orange is being studied. Once bound, the fluorescence of the ligand is enhanced up to 1200 fold with a detection limit of in the pmol range. The use of this aptamer for use in a lateral flow assay is being explored due to the high signal to noise ratio conferred by this system. The kinetics of the binding will be studied using the Open SPR system.
SPR is a powerful tool that can be used to study the binding kinetics of biomolecules in a real-time fashion. The malachite green aptamer is used as a model system to explore aptamer binding to its ligand that is immobilized on a gold surface. It has been shown that the kinetics of the binding interaction is greatly affected by the identity and concentration of the positive ions in solution through isothermal titration calorimetry.
1. Dolgosheina E V, Jeng SCY, Panchapakesan SSS, Cojocaru R, Chen PSK, Wilson PD, Hawkins N, Wiggins PA, Unrau PJ: RNA Mango Aptamer-Fluorophore: A Bright, High-Affinity Complex for RNA Labeling and Tracking. ACS Chem. Biol. 2014, 9:2412–2420.
Da Costa JB, Andreiev AI, Dieckmann T: Thermodynamics and Kinetics of Adaptive Binding in the Malachite Green RNA Aptamer. Biochemistry 2013, 52:6575–6583.
Bernard Da Costa J, Dieckmann T: Entropy and Mg(2+) control ligand affinity and specificity in the malachite green binding RNA aptamer. Mol Biosyst 2011, 7:2156–2163.
Keywords: Aptamer, Lateral flow, SPR