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Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection

Barbara W Sódar1, Ágnes Kittel2, Krisztina Pálóczi1, Krisztina V Vukman1,
Xabier Osteikoetxea1, Katalin Szabó-Taylor1, Andrea Németh1, Beáta Sperlágh2,
Tamás Baranyai3, Zoltán Giricz3, Zoltán Wiener1, Lilla Turiák4, László Drahos4, Éva Pállinger1,
Károly Vékey4, Péter Ferdinandy3, András Falus1 & Edit Irén Buzás

Published: 18 April 2016 by Scientific Reports

Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of
diseases. Despite the increasing interest, their isolation and purification from body fluids remains
challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma
samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of
circulating particles within the size range of extracellular vesicles lacked common vesicular markers.
We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL)
which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on
biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular
vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from
blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed
an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show copurification
and in vitro association of LDL with extracellular vesicles and its interference with vesicle
analysis. Our data point to the importance of careful study design and data interpretation in studies
using blood-derived extracellular vesicles with special focus on potentially co-purified LDL.

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