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024 7 _ |a 10.3390/ijms22179211
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024 7 _ |a pmc:PMC8431127
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024 7 _ |a 1422-0067
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024 7 _ |a 1661-6596
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037 _ _ |a DZNE-2021-01466
041 _ _ |a English
082 _ _ |a 540
100 1 _ |a Tzaridis, Theophilos
|0 0000-0001-9651-1144
|b 0
245 _ _ |a Extracellular Vesicle Separation Techniques Impact Results from Human Blood Samples: Considerations for Diagnostic Applications.
260 _ _ |a Basel
|c 2021
|b Molecular Diversity Preservation International
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500 _ _ |a CC BY
520 _ _ |a 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.
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650 _ 7 |a extracellular vesicle isolation
|2 Other
650 _ 7 |a extracellular vesicles diagnostics
|2 Other
650 _ 7 |a methods in liquid biopsy
|2 Other
650 _ 7 |a plasma biomarker
|2 Other
650 _ 7 |a serum biomarker
|2 Other
650 _ 7 |a Biomarkers
|2 NLM Chemicals
650 _ 7 |a Proteins
|2 NLM Chemicals
650 _ 2 |a Biological Transport
|2 MeSH
650 _ 2 |a Biomarkers
|2 MeSH
650 _ 2 |a Cell Fractionation: methods
|2 MeSH
650 _ 2 |a Chemical Fractionation: methods
|2 MeSH
650 _ 2 |a Extracellular Vesicles: metabolism
|2 MeSH
650 _ 2 |a Extracellular Vesicles: ultrastructure
|2 MeSH
650 _ 2 |a Flow Cytometry
|2 MeSH
650 _ 2 |a Humans
|2 MeSH
650 _ 2 |a Liquid Biopsy: methods
|2 MeSH
650 _ 2 |a Proteins: metabolism
|2 MeSH
700 1 _ |a Bachurski, Daniel
|0 0000-0001-9168-9680
|b 1
700 1 _ |a Liu, Shu
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700 1 _ |a Surmann, Kristin
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700 1 _ |a Babatz, Felix
|b 4
700 1 _ |a Gesell Salazar, Manuela
|b 5
700 1 _ |a Völker, Uwe
|b 6
700 1 _ |a Hallek, Michael
|b 7
700 1 _ |a Herrlinger, Ulrich
|b 8
700 1 _ |a Vorberg, Ina
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700 1 _ |a Coch, Christoph
|b 10
700 1 _ |a Reiners, Katrin S
|0 0000-0002-5908-8823
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700 1 _ |a Hartmann, Gunther
|b 12
770 _ _ |a Circulating Molecules and Precision Medicine in Cancer
773 _ _ |a 10.3390/ijms22179211
|g Vol. 22, no. 17, p. 9211 -
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|p 9211
|t International journal of molecular sciences
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856 4 _ |y OpenAccess
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