Luminescent Terbium Probe for Time-Resolved FRET and NSET Binding Assays with Quantum Dots and Gold Nanoparticles

Abstract

Time-gated or time-resolved FRET (TR-FRET) assays are important tools in biosensing, bioimaging, drug screening, and molecular diagnostics. Efficient TR-FRET assays require stable lanthanide complexes with high absorption cross sections, high quantum yields, and long photoluminescence lifetimes. Owing to their challenging synthesis, such complexes are relatively rare and new components are of potential interest when developing TR-FRET probes. Here, we evaluate the recently developed Tb complex CoraFluor-1 concerning its analytical performance in terbium-to-quantum dot FRET and terbium-to-gold nanoparticle NSET assays using the prototypical biological recognition system of streptavidin and biotin. Biological binding was quantifiable at sub-picomolar concentrations in small sample volumes, with broad applicability demonstrated across three commercial fluorescence plate readers used for time-resolved, spectrally-resolved, and clinical bioanalysis. Overall, CoraFluor-1 provided excellent analytical performance as both FRET and NSET donor, validating its potential for developing new TR-FRET probes for biosensing and bioimaging.