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

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 sa...

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Published inScientific reports Vol. 6; no. 1; p. 24316
Main Authors Sódar, Barbara W, Kittel, Ágnes, Pálóczi, Krisztina, Vukman, Krisztina V, Osteikoetxea, Xabier, Szabó-Taylor, Katalin, Németh, Andrea, Sperlágh, Beáta, Baranyai, Tamás, Giricz, Zoltán, Wiener, Zoltán, Turiák, Lilla, Drahos, László, Pállinger, Éva, Vékey, Károly, Ferdinandy, Péter, Falus, András, Buzás, Edit Irén
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group UK 18.04.2016
Nature Publishing Group
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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.
AbstractList 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.
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.
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.
ArticleNumber 24316
Author Falus, András
Pállinger, Éva
Buzás, Edit Irén
Vukman, Krisztina V
Drahos, László
Giricz, Zoltán
Vékey, Károly
Szabó-Taylor, Katalin
Pálóczi, Krisztina
Wiener, Zoltán
Sperlágh, Beáta
Ferdinandy, Péter
Németh, Andrea
Turiák, Lilla
Kittel, Ágnes
Sódar, Barbara W
Osteikoetxea, Xabier
Baranyai, Tamás
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  organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University
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  organization: Research Centre for Natural Sciences, Hungarian Academy of Sciences
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  organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University
BackLink https://www.ncbi.nlm.nih.gov/pubmed/27087061$$D View this record in MEDLINE/PubMed
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Snippet Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and...
Abstract Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their...
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proquest
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springer
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Index Database
Publisher
StartPage 24316
SubjectTerms 14/28
631/1647/1407/1492
631/1647/2230
82/16
82/58
Adult
Biomarkers - blood
Blood
Blood platelets
Blood Platelets - chemistry
Body fluids
Data interpretation
Data processing
Electron microscopy
Exosomes
Exosomes - chemistry
Extracellular Vesicles - chemistry
Female
Flow cytometry
Humanities and Social Sciences
Humans
Lipoproteins
Lipoproteins, LDL - blood
Low density lipoprotein
Male
multidisciplinary
Postprandial Period
Purification
Science
Transmission electron microscopy
Vesicles
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  providerName: Springer Nature
Title Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection
URI https://link.springer.com/article/10.1038/srep24316
https://www.ncbi.nlm.nih.gov/pubmed/27087061
https://www.proquest.com/docview/1898680797
https://search.proquest.com/docview/1782217793
https://pubmed.ncbi.nlm.nih.gov/PMC4834552
Volume 6
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