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

Nat Biomed Eng. 2020 May;4(5):518-530. doi: 10.1038/s41551-020-0547-4. Epub 2020 Apr 20.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biological Assay / methods*
  • Bone Marrow Cells / cytology
  • Cell Line, Tumor
  • Colloids / chemistry
  • Dendritic Cells / cytology
  • Female
  • Flow Cytometry
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Humans
  • Immunoassay
  • Lipopolysaccharides / pharmacology
  • Mice, Inbred C57BL
  • Microspheres
  • Nanoparticles / chemistry*
  • Proteomics
  • Reference Standards

Substances

  • Colloids
  • Fluorescent Dyes
  • Lipopolysaccharides

Associated data

  • figshare/10.6084/m9.figshare.11888055