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@ARTICLE{Tzaridis:162811,
author = {Tzaridis, Theophilos and Bachurski, Daniel and Liu, Shu and
Surmann, Kristin and Babatz, Felix and Gesell Salazar,
Manuela and Völker, Uwe and Hallek, Michael and Herrlinger,
Ulrich and Vorberg, Ina and Coch, Christoph and Reiners,
Katrin S and Hartmann, Gunther},
title = {{E}xtracellular {V}esicle {S}eparation {T}echniques
{I}mpact {R}esults from {H}uman {B}lood {S}amples:
{C}onsiderations for {D}iagnostic {A}pplications.},
journal = {International journal of molecular sciences},
volume = {22},
number = {17},
issn = {1422-0067},
address = {Basel},
publisher = {Molecular Diversity Preservation International},
reportid = {DZNE-2021-01466},
pages = {9211},
year = {2021},
note = {CC BY},
abstract = {Extracellular vesicles (EVs) are reminiscent of their cell
of origin and thus represent a valuable source of
biomarkers. However, for EVs to be used as biomarkers in
clinical practice, simple, comparable, and reproducible
analytical methods must be applied. Although progress is
being made in EV separation methods for human biofluids, the
implementation of EV assays for clinical diagnosis and
common guidelines are still lacking. We conducted a
comprehensive analysis of established EV separation
techniques from human serum and plasma, including
ultracentrifugation and size exclusion chromatography (SEC),
followed by concentration using (a) ultracentrifugation, (b)
ultrafiltration, or (c) precipitation, and immunoaffinity
isolation. We analyzed the size, number, protein, and miRNA
content of the obtained EVs and assessed the functional
delivery of EV cargo. Our results demonstrate that all
methods led to an adequate yield of small EVs. While no
significant difference in miRNA content was observed for the
different separation methods, ultracentrifugation was best
for subsequent flow cytometry analysis. Immunoaffinity
isolation is not suitable for subsequent protein analyses.
SEC + ultracentrifugation showed the best functional
delivery of EV cargo. In summary, combining SEC with
ultracentrifugation gives the highest yield of pure and
functional EVs and allows reliable analysis of both protein
and miRNA contents. We propose this combination as the
preferred EV isolation method for biomarker studies from
human serum or plasma.},
keywords = {Biological Transport / Biomarkers / Cell Fractionation:
methods / Chemical Fractionation: methods / Extracellular
Vesicles: metabolism / Extracellular Vesicles:
ultrastructure / Flow Cytometry / Humans / Liquid Biopsy:
methods / Proteins: metabolism / extracellular vesicle
isolation (Other) / extracellular vesicles diagnostics
(Other) / methods in liquid biopsy (Other) / plasma
biomarker (Other) / serum biomarker (Other) / Biomarkers
(NLM Chemicals) / Proteins (NLM Chemicals)},
cin = {AG Vorberg},
ddc = {540},
cid = {I:(DE-2719)1013004},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:34502122},
pmc = {pmc:PMC8431127},
doi = {10.3390/ijms22179211},
url = {https://pub.dzne.de/record/162811},
}