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Dual-Wavelength Surface Plasmon Resonance for Determining the Size and Concentration of Sub-Populations of Extracellular Vesicles

Journal article
Authors Déborah L. M. Rupert
Ganesh V Shelke
Gustav Emilsson
Virginia Claudio
Stephan Block
Cecilia Lässer
Andreas Dahlin
Jan Lötvall
Marta Bally
Vladimir P. Zhdanov
Fredrik Höök
Published in Analytical Chemistry
Volume 88
Issue 20
Pages 9980-9988
ISSN 0003-2700
Publication year 2016
Published at Krefting Research Centre
Pages 9980-9988
Language en
Keywords Dual-Wavelength Surface Plasmon Resonance, Extracellular Vesicles
Subject categories Cell Biology, Nano Technology


Accurate concentration determination of subpopulations of extracellular vesicles (EVs), such as exosomes, is of importance both in the context of understanding their fundamental biological role and of potentially using them as disease biomarkers. In principle, this can be achieved by measuring the rate of diffusion-limited mass uptake to a sensor surface modified with a receptor designed to only bind the subpopulation of interest. However, a significant error is introduced if the targeted EV subpopulation has a size, and thus hydrodynamic diffusion coefficient, that differs from the mean size and diffusion coefficient of the whole EV population and/or if the EVs become deformed upon binding to the surface. We here demonstrate a new approach to determine the mean size (or effective film thickness) of bound nanoparticles, in general, and EV subpopulation carrying a marker of interest, in particular. The method is based on operating surface plasmon resonance simultaneously at two wavelengths with different sensing depths and using the ratio of the corresponding responses to extract the particle size on the surface. By estimating in this way the degree of deformation of adsorbed EVs, we markedly improved their bulk concentration determination and showed that EVs carrying the exosomal marker CD63 correspond to not more than around 10% of the EV sample.

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