Understanding FRET in Upconversion Nanoparticle Nucleic Acid Biosensors.

Upconversion nanoparticles (UCNPs) have been frequently applied in Förster resonance energy transfer (FRET) bioanalysis. However, the understanding of how surface coatings, bioconjugation, and dye-surface distance influence FRET biosensing performance has not significantly advanced. Here, we investigated UCNP-to-dye FRET DNA-hybridization assays in HO and DO using ∼24 nm large NaYF:Yb,Er UCNPs coated with thin layers of silica (SiO) or poly(acrylic acid) (PAA).

Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET.

Time-gated Förster resonance energy transfer (TG-FRET) between Tb complexes and luminescent semiconductor quantum dots (QDs) provides highly advantageous photophysical properties for multiplexed biosensing. Multiplexed Tb-to-QD FRET immunoassays possess a large potential for in vitro diagnostics, but their performance is often insufficient for their application under clinical conditions. Here, we developed a homogeneous TG-FRET immunoassay for the quantification of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and prostate-specific antigen (PSA) from a single serum sample by multiplexed Tb-to-QD FRET.

In Vivo Biosensing Using Resonance Energy Transfer.

Solution-phase and intracellular biosensing has substantially enhanced our understanding of molecular processes foundational to biology and pathology. Optical methods are favored because of the low cost of probes and instrumentation. While chromatographic methods are helpful, fluorescent biosensing further increases sensitivity and can be more effective in complex media.

Sensing with photoluminescent semiconductor quantum dots.

Fluorescent sensors benefit from high signal-to-noise and multiple measurement modalities, enabling a multitude of applications and flexibility of design. Semiconductor nanocrystal quantum dots (QDs) are excellent fluorophores for sensors because of their extraordinary optical properties. They have high thermal and photochemical stability compared to organic dyes or fluorescent proteins and are extremely bright due to their large molar cross-sections.

Direct conjugation of antibodies to the ZnS shell of quantum dots for FRET immunoassays with low picomolar detection limits.

Compact and functional nanoparticle-antibody conjugates are of paramount importance for the development of quantum dot (QD)-based immunoassays. Here, we present a simple strategy to directly conjugate IgG, F(ab'), and Fab antibodies via their endogenous disulfide groups directly to the inorganic ZnS shell of compact penicillamine-coated QDs. The functionality of the conjugates was demonstrated by terbium (Tb)-to-QD FRET immunoassays against prostate specific antigen in serum samples.