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

Déborah L. M. Rupert (Institutionen för fysik, Biologisk fysik (Chalmers)) ; Ganesh V Shelke ; Gustav Emilsson (Institutionen för fysik, Bionanofotonik (Chalmers)) ; Virginia Claudio (Institutionen för fysik, Biologisk fysik (Chalmers)) ; Stephan Block (Institutionen för fysik, Biologisk fysik (Chalmers)) ; Cecilia Lässer ; Andreas Dahlin (Institutionen för fysik, Bionanofotonik (Chalmers)) ; Jan Lötvall ; Marta Bally (Institutionen för fysik, Biologisk fysik (Chalmers)) ; Vladimir P. Zhdanov (Institutionen för fysik, Kemisk fysik (Chalmers)) ; Fredrik Höök (Institutionen för fysik, Biologisk fysik (Chalmers))
Analytical Chemistry (0003-2700). Vol. 88 (2016), 20, p. 9980-9988.
[Artikel, refereegranskad vetenskaplig]

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.

Nyckelord: Dual-Wavelength Surface Plasmon Resonance, Extracellular Vesicles

Denna post skapades 2016-10-14. Senast ändrad 2017-10-03.
CPL Pubid: 243377


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