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Quantifying protein aggregate concentration through electrochemical oxidation of thioflavin T
Narvydas Dėnas1, Povilas Virbickas2, Aušra Valiūnienė1
1Institute of Chemistry, Faculty of Chemistry and Geosciences, Vilnius University, Vilnius, Lithuania; State Research Institute Center for Physical Sciences and Technology, Sauletekio Ave. 3, LT-10257 Vilnius, Lithuania.
None:
Investigating protein aggregation and determining the concentration of protein aggregates (PAs) is important in clinical studies and in the food industry. However, conventional fluorimetric methods that use amyloidophilic dyes are limited by comparatively high cost and interference from light scattering and other optically active contaminants, which can affect the reliability of measurements. This study presents an electrochemical approach to quantifying the concentration of a model PA, lysozyme aggregate (LA), by measuring the oxidation current of Thioflavin T (ThT), which LA molecules absorb. Results demonstrate that cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) provide linear detection ranges of ThT of 140 μM-1220 μM and 26 μM-170 μM, respectively. Additionally, CV studies in buffer solutions containing ThT and LA reveal that ThT-LA interaction diminishes the ThT oxidation current, enabling the detection of LA concentrations ranging from 3 μg mL-1 to 1190 μg mL-1. The optimal ThT concentration for electrochemical LA detection is approximately 1 mM. These findings suggest that using ThT for electrochemical sensing of protein aggregates offers a promising alternative to fluorimetry.
