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Bioassays for the rapid detection and quantification of specific nucleic acids, proteins, and peptides are fundamental tools in many clinical settings. Traditional optical emission methods have focused on the use of molecular dyes as labels to track selective binding interactions and as probes that are sensitive to environmental changes. Such dyes can offer good detection limits based on brightness but typically have broad emission bands and suffer from time-dependent photobleaching. Inorganic nanoparticles such as quantum dots and upconversion nanoparticles are photostable over prolonged exposure to excitation radiation and tend to offer narrow emission bands, providing a greater opportunity for multiwavelength multiplexing. Importantly, in contrast to molecular dyes, nanoparticles offer substantial surface area and can serve as platforms to carry a large number of conjugated molecules. The surface chemistry of inorganic nanoparticles offers both challenges and opportunities for the control of solubility and functionality for selective molecular interactions by the assembly of coatings through coordination chemistry. This report reviews advances in the compositional design and methods of conjugation of inorganic quantum dots and upconversion nanoparticles and the assembly of combinations of nanoparticles to achieve energy exchange. Our interest is the exploration of configurations where the modified nanoparticles can be immobilized to solid substrates for the development of bioassays and biosensors that operate by resonance energy transfer (RET).

Monodisperse water-soluble upconverting nanoparticles (UCNPs) were immobilized onto modified glass substrates for development of biosensing surfaces that operated using luminescence resonance energy transfer (LRET). Amine modified UCNPs were prepared from oleic acid capped UCNPs by ligand exchange using o-phosphorylethanolamine (PEA). PEA-UCNPs were covalently immobilized on aldehyde functionalized coverslips. Environmental scanning electron microscopy (ESEM) images indicated a homogeneous distribution of UCNPs on surfaces with a high immobilization density of approximately 1.3 × 10(11) UCNP cm(-2). This is the first account of covalent immobilization of UCNPs for bioassay and biosensor development where the density is on par with the high immobilization densities reported for other types of nanoparticles. The functionality and stability of the immobilized NPs were demonstrated by examining an LRET-based bioassay. The well-known sandwich assay for the detection of thrombin was selected as a model in which UCNPs were used as donors and quantum dots (QDs) as acceptors. The closely packed UCNPs on the glass surface showed a 2.5-fold enhancement in assay sensitivity compared to less-densely packed surfaces. In addition, a 1.5-fold enhancement in energy transfer efficiency was shown for solid-phase compared to solution-phase LRET.

Targeted drug delivery using functional nanoparticles has provided new strategies for improving therapeutic efficacy while concurrently minimizing toxicity. Photodynamic therapy is an approach that offers control of drug delivery by use of an external photon source to allow active therapeutic release to a target area. Upconverting nanoparticles (UCNPs) have potential to operate as integral components of photodynamic therapeutic platforms based on the resonant absorption of near-infrared (NIR) radiation and emission at shorter wavelengths. NIR radiation is minimally absorbed and scattered by biological tissues, and the NIR excitation of UCNPs can generate anti-Stokes emission in the ultraviolet-visible wavelength range at intensities that can be used to trigger cleavage of bonds linking therapeutics at the nanoparticle interface. Herein, we describe an investigation of photocleavage at the surface of UCNPs to release the chemotherapeutic 5-fluorouracil (5-FU). Core-shell UCNPs composed of a β-NaYF4: 4.95% Yb, 0.08% Tm core and a β-NaYF4 shell were coated with o-phosphorylethanolamine ligands and coupled to an o-nitrobenzyl (ONB) derivative of 5-FU. NIR excitation of the UCNPs resulted in photoluminescence (PL) emission bands centered at 365, 455, and 485 nm. The UV-blue PL was in resonance with the absorption band of the ONB-FU derivative resulting in photocleavage and subsequent release of the 5-FU drug from the UCNPs for these in vitro studies. The release of 5-FU was complete in <14 min using a NIR laser source centered at 980 nm that operated at a power of <100 mW. The efficiency of triggered release was as high as 77% of the total ONB-FU conjugate, while the rate of drug release could be tuned with the laser power output. This work provides an important first step in the development of a UCNP platform capable of targeted chemotherapy.