Assembly of single-stranded polydeoxyadenylic acid and β-glucan probed by the sensing platform of graphene oxide based on the fluorescence resonance energy transfer and fluorescence anisotropy

Abstract

Based on the fluorescence resonance energy transfer (FRET) and fluorescence anisotropy (FA), the present study reported proof-of-principle for a highly sensitive and rapid detection technique that can be precisely utilized for investigating the self-assembly of polydeoxyadenylic acid (poly(dA)) and β-glucan, and the interactions of the poly(dA)–β-glucan complex on the surface of graphene oxide (GO). Due to the noncovalent assembly of fluorescein amidite (FAM)-labeled poly(dA) and GO via π–π stacking, the fluorescence of (FAM)-labeled poly(dA) as a molecular aptamer beacon (MAB) was completely quenched by GO. Conversely, the addition of single-stranded lentinan (s-LNT) resulted in the significant restoration of fluorescence due to the formation of poly(dA)–s-LNT complexes with a stiff rod-like structure, which had a weak affinity to GO and kept the dyes away from GO. However, relatively weak fluorescence restoration was observed by adding another single-stranded curdlan (s-CUR) for positive control, indicative of complex formation with higher binding ability to GO. The fluorescence anisotropy (FA) was also combined to confirm the occurrence with different increments of anisotropy relative to the free poly(dA), which could be conveniently extended for detecting the assembly of other biomolecules with higher sensitivity.