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Physical characterization of exosomes and other extracellular vesicles using surface-based sensing

Déborah L. M. Rupert (Institutionen för teknisk fysik, Biologisk fysik)
Göteborg : Chalmers University of Technology, 2015. ISBN: 978-91-7597-291-6.- 68 s.
[Doktorsavhandling]

Extracellular vesicles (EVs) are hundred of nanometer sized particles secreted by almost all cell types. Exosomes are one type of EVs today intensively investigated due to the increased evidence suggesting that they play important roles as communicating vehicles in a broad range of physiological processes, pathological conditions and disease diagnostics. Their multifaceted profiles highlight the importance of elucidating how their different modes of biological action correlate with their biomolecular profiles, physicochemical properties and their concentration in biological fluids. This thesis explores the potential of two surface-based sensor methods, namely surface-plasmon resonance (SPR) and optical waveguide scattering / fluorescence microscopy (OWSFM), for physical characterization of EVs. EVs carrying specific exosomal marker were quantified in terms of bulk concentration using SPR. A key uncertainty of this quantification approach was suggested to originate from vesicles contraction upon adsorption to the sensor surface. This concern was scrutinized with dual-wavelength SPR, illustrating the advantage of dual-wavelength SPR over conventional SPR to quantify nanoparticle contraction in general and concentration determination of EVs in particular. As an extension of these findings, OWSFM was utilized to compare the optical density and lipid content of two sub-populations of extracellular vesicles observed at the single particle level in both scattering and fluorescence mode. In summary, the thesis work contributes with new tools for improved characterization of physicochemical properties of different EVs and concentration determinations of sub-population of EVs carrying exosomal markers.

Nyckelord: Extracellular vesicles, exosomes, liposomes, surface-based biosensing, cluster of differentiation 63 (CD63), antibody, specificity, single particle, label-free, optical waveguide microscopy, surface plasmon resonance, nanoparticle tracking analysis, scattering, fluorescence, optical density, lipid content, quantification, size distribution



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Denna post skapades 2015-11-16. Senast ändrad 2015-11-17.
CPL Pubid: 225809

 

Institutioner (Chalmers)

Institutionen för teknisk fysik, Biologisk fysik (2007-2015)

Ämnesområden

Materialvetenskap
Nanovetenskap och nanoteknik
Analytisk kemi
Biofysikalisk kemi
Biofysik

Chalmers infrastruktur

Examination

Datum: 2015-12-15
Tid: 13:00
Lokal: Kollektorn, MC2, Chalmers Tekniska Högskola, Kemivägen 9, 41296 Göteborg,
Opponent: Janos Vörös, Professor, Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, Department for Information Technology and Electrical Engineering, Swiss Federal Institute of Technology in Zurich (ETH Zurich), Switzerland.