Electrochemical sandwich immunosensor for determination of exosomes based on surface marker-mediated signal amplification

exosomes

Extracellular vesicles (EVs), namely exosomes and microvesicles, are important mediators of intercellular communication pathways. Since EVs can be detected in a variety of biofluids and contain a specific set of biomarkers which are reminiscent of their parental cells, they show great promise in clinical diagnostics as EV analysis can be performed in minimally-invasive liquid biopsies. However, reliable, fast and cost-effective methods for their determination are still needed, especially if decentralized analysis is intended.

In this study, researchers at the Universidad de la República, Uruguay developed an electrochemical biosensor which works with 1.5 µL sample volume, and can detect as low as 200 exosomes per microliter, with a linear range spanning almost four orders of magnitude. The sensor is specific, and readily differentiates exosomes from microvesicles in samples containing 1000-fold excess of the latter. Capability of detecting exosomes in real samples (diluted serum) was shown.This was achieved by immobilizing rabbit anti-human CD9 antibodies on gold substrates, and using monoclonal antibodies against CD9 for detection of captured exosomes. Signal amplification is presumably obtained from the fact that multiple detector antibodies bind to the surface of each captured vesicle. Detection is performed based on electrochemical reduction of 3, 3′, 5, 5′-tetramethyl benzidine (TMB) after addition of horseradish peroxidase (HRP)-conjugated anti-IgG antibodies. This amperometric biosensor can be easily incorporated into future miniaturized and semi-automatic devices for EV determination.

Doldán X, Fagúndez P, Cayota A, Laíz J, Tosar JP. (2016) Electrochemical sandwich immunosensor for determination of exosomes based on surface marker-mediated signal amplification. Anal Chem [Epub ahead of print]. [abstract]

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