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

超亮荧光纳米级标签可用于标准生物测定法对飞摩尔级分析物进行检测

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作者:Jingyi Luan, Anushree Seth, Rohit Gupta, Zheyu Wang, Priya Rathi, Sisi Cao, Hamed Gholami Derami, Rui Tang, Baogang Xu, Samuel Achilefu, Jeremiah J Morrissey, Srikanth Singamaneni

Abstract

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 readout devices. The plasmonic construct consists of a bovine serum albumin scaffold with approximately 210 IRDye 800CW fluorophores (with a fluorescence intensity approximately 6,700-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 improves the limit of detection in fluorescence-linked immunosorbent assays by up to 4,750-fold and is compatible with multiplexed bead-based immunoassays, immunomicroarrays, flow cytometry and immunocytochemistry methods, and it shortens overall assay times (to 20 min) 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.

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