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

Sódar, Barbara W and Kittel, Ágnes and Pálóczi, Krisztina and Vukman, Krisztina V and Osteikoetxea, Xabier and Szabó-Taylor, Katalin and Németh, Andrea and Sperlágh, Beáta and Baranyai, Tamás and Giricz, Zoltán and Wiener, Zoltán and Turiák, Lilla and Drahos, László and Pállinger, Éva and Vékey, Károly and Ferdinandy, Péter and Falus, András and Buzás, Edit Irén (2016) Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection. SCIENTIFIC REPORTS, 6. pp. 1-12. ISSN 2045-2322

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Abstract

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 co-purification 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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