Ultrabright fluorescent nanoscale labels for the femtomolar detection of analytes with standard bioassays

The detection and quantification of low-abundance molecular biomarkers in biological samples is challenging. Here, we show that a plasmonic nanoscale construct serving as an ‘add-on’ label for a broad range of bioassays improves their signal-to-noise ratio and dynamic range without altering their workflow and read-out devices. The plasmonic construct consists of a bovine-serum-albumin scaffold with approximately 210 IRDye 800CW fluorophores (with fluorescence intensity approximately 6700-fold that of a single 800CW fluorophore), a polymer-coated gold nanorod acting as a plasmonic antenna, and biotin as a high-affinity biorecognition element. Its emission wavelength can be tuned over the visible and near-infrared spectral regions by modifying its size, shape and composition. It is compatible with multiplexed bead-based immunoassays (it improves the limit of detection by up to 4,750-fold in fluorescence-linked immunosorbent assays), immuno-microarrays, flow-cytometry and immunocytochemistry methods, and it shortens overall assay times and lowers sample volumes, as shown for the detection of a pro-inflammatory cytokine in mouse interstitial fluid and of urinary biomarkers in patient samples.