Self-Assembled Nanoparticle Probes for Recognition and Detection of Biomolecules

TLDR

The biosensor uses a 2.5‑nm gold nanoparticle as scaffold and quencher, with thiol‑labeled oligonucleotides bearing fluorophores that, upon target binding, undergo a conformational change that restores fluorescence. The resulting nanoparticle probes form a constrained arch‑like structure, detect specific DNA sequences and single‑base mutations in a homogeneous assay, exhibit minimal temperature‑dependent background fluorescence, and outperform conventional organic quenchers such as Dabcyl.

Abstract

Colloidal gold nanocrystals have been used to develop a new class of nanobiosensors that is able to recognize and detect specific DNA sequences and single-base mutations in a homogeneous format. At the core of this biosensor is a 2.5-nm gold nanoparticle that functions as both a nano-scaffold and a nano-quencher (efficient energy acceptor). Attached to this core are oligonucleotide molecules labeled with a thiol group at one end and a fluorophore at the other. This hybrid bio/inorganic construct is found to spontaneously assemble into a constrained arch-like conformation on the particle surface. Binding of target molecules results in a conformational change, which restores the fluorescence of the quenched fluorophore. Unlike conventional molecular beacons with a stem-and-loop structure, the nanoparticle probes do not require a stem, and their background fluorescence increases little with temperature. In comparison with the organic quencher Dabcyl (4,4‘-dimethylaminophenyl azo benzoic acid), metal nanoparticles have unique structural and optical properties for new applications in biosensing and molecular engineering.