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Issue 43, 2018
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A dual-signal amplification platform for sensitive fluorescence biosensing of leukemia-derived exosomes

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Abstract

Exosomes as nanosized biomarkers hold great potential for the diagnosis of cancer. However, the low concentration of cancer-derived exosomes present in biofluids makes early diagnosis strenuous. Here, we developed a fluorescent biosensing platform, namely a dual signal amplification, for the ultrasensitive detection of leukemia cell-derived exosomes. The protocol consists of three steps: first, leukemia-derived exosomes containing CD63 and nucleolin were captured by anti-CD63 antibody modified magnetic bead conjugates (MB-CD63); then, a DNA primer comprising a nucleolin-recognition aptamer (AS1411) was applied to bind the exosomes which further initiated a rolling circle amplification (RCA) reaction to generate many repeat sequences for hybridization with gold nanoparticle (GNP)–DNA–fluorescent dye (FAM) conjugates (GNP–DNA–FAM); finally, nicking endonuclease (Nb·BbvCI) assisted target recycling was introduced. As a result, FAM was released from GNP–DNA–FAM conjugates, transformed from the quenching state to the emission state and thus fluorescence signals continuously accumulated. With this dual signal amplification platform, as low as 1 × 102 particles per μL exosomes could be detected. Furthermore, we have successfully applied this method for the detection of exosomes in spiked serum samples, indicating a promising tool for clinical application.

Graphical abstract: A dual-signal amplification platform for sensitive fluorescence biosensing of leukemia-derived exosomes

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Supplementary files

Article information


Submitted
22 Sep 2018
Accepted
02 Oct 2018
First published
23 Oct 2018

This article is Open Access

Nanoscale, 2018,10, 20289-20295
Article type
Paper

A dual-signal amplification platform for sensitive fluorescence biosensing of leukemia-derived exosomes

L. Huang, D. Wang, N. Singh, F. Yang, N. Gu and X. Zhang, Nanoscale, 2018, 10, 20289
DOI: 10.1039/C8NR07720G

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    [Original citation] - Published by The Royal Society of Chemistry.

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