TY - JOUR
T1 - Ultrabright fluorescent nanoscale labels for the femtomolar detection of analytes with standard bioassays
AU - Luan, Jingyi
AU - Seth, Anushree
AU - Gupta, Rohit
AU - Wang, Zheyu
AU - Rathi, Priya
AU - Cao, Sisi
AU - Gholami Derami, Hamed
AU - Tang, Rui
AU - Xu, Baogang
AU - Achilefu, Samuel
AU - Morrissey, Jeremiah J.
AU - Singamaneni, Srikanth
N1 - Funding Information:
We acknowledge support from National Science Foundation (award nos. CBET-1512043 and CBET-1900277), National Institutes of Health (R01DE027098 and R01CA141521), Center for Multiple Myeloma Nanotherapy (U54 CA199092) and a grant from the Barnes-Jewish Hospital Research Foundation (no. 3706). We thank K. Naegle for providing access to a LI-COR Odyssey CLx scanner, L. Setton for the flow cytometer, J. Rudra and T. Pietka for Luminex readers, and the Nano Research Facility (NRF) and Institute of Materials Science and Engineering (IMSE) at Washington University for providing access to electron microscopy facilities. We also thank G. Genin for inspiring discussions and suggestions.
Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2020/5/1
Y1 - 2020/5/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85083741540&partnerID=8YFLogxK
U2 - 10.1038/s41551-020-0547-4
DO - 10.1038/s41551-020-0547-4
M3 - Article
C2 - 32313101
AN - SCOPUS:85083741540
SN - 2157-846X
VL - 4
SP - 518
EP - 530
JO - Nature Biomedical Engineering
JF - Nature Biomedical Engineering
IS - 5
ER -