Monodisperse water-soluble upconverting nanoparticles (UCNPs) were immobilized onto paper for the development of a triplex optical-based assay that operated using luminescence resonance energy transfer (LRET).  Multiplexing, the simultaneous detection of multiple biomolecules, is important for accurate and rapid determinations in medical diagnostics and biotechnology. Herein, we use blue-emitting UCNPs as donors and different colour quantum dots (QDs) as acceptors to achieve a high multiplexing capacity in LRET owing to the narrow and tunable emission bands of both nanoparticles. The assay was developed for the definitive diagnosis of hyperprothrombinemia, the overexpression of thrombin, which increases the risk of thrombosis. Housekeeping DNA HPRT found in hepatocyte cells, an mRNA fragment and its expressed protein, thrombin, were detected. DNA probes for the detection of DNA and mRNA fragments in addition to an aptamer for the detection of thrombin were simultaneously immobilized on the surface of the UCNPs. Each captured biomolecule was then labeled by selective interactions in a sandwich format with QDs decorated with respective reporters. The intensity of LRET emission from each type of QD served to identify and quantify the three targets allowing for multiplexing with one excitation source (NIR) and one donor (UCNP). The detection of housekeeping genes helps identify cell lines, while mRNA and protein levels serve as dual parameters for diagnosis. While thrombin is used as a prototype here, high levels of proteins are common in many cancers where this assay design can find new applications.

Upconverting nanoparticles in solid-phase luminescence resonance energy transfer based assays for the high capacity multiplexed detection of biomolecules

249th National Meeting (2015)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.