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000162811 037__ $$aDZNE-2021-01466
000162811 041__ $$aEnglish
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000162811 1001_ $$00000-0001-9651-1144$$aTzaridis, Theophilos$$b0
000162811 245__ $$aExtracellular Vesicle Separation Techniques Impact Results from Human Blood Samples: Considerations for Diagnostic Applications.
000162811 260__ $$aBasel$$bMolecular Diversity Preservation International$$c2021
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000162811 520__ $$aExtracellular 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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000162811 650_7 $$2Other$$aextracellular vesicle isolation
000162811 650_7 $$2Other$$aextracellular vesicles diagnostics
000162811 650_7 $$2Other$$amethods in liquid biopsy
000162811 650_7 $$2Other$$aplasma biomarker
000162811 650_7 $$2Other$$aserum biomarker
000162811 650_7 $$2NLM Chemicals$$aBiomarkers
000162811 650_7 $$2NLM Chemicals$$aProteins
000162811 650_2 $$2MeSH$$aBiological Transport
000162811 650_2 $$2MeSH$$aBiomarkers
000162811 650_2 $$2MeSH$$aCell Fractionation: methods
000162811 650_2 $$2MeSH$$aChemical Fractionation: methods
000162811 650_2 $$2MeSH$$aExtracellular Vesicles: metabolism
000162811 650_2 $$2MeSH$$aExtracellular Vesicles: ultrastructure
000162811 650_2 $$2MeSH$$aFlow Cytometry
000162811 650_2 $$2MeSH$$aHumans
000162811 650_2 $$2MeSH$$aLiquid Biopsy: methods
000162811 650_2 $$2MeSH$$aProteins: metabolism
000162811 7001_ $$00000-0001-9168-9680$$aBachurski, Daniel$$b1
000162811 7001_ $$0P:(DE-2719)2810461$$aLiu, Shu$$b2$$udzne
000162811 7001_ $$aSurmann, Kristin$$b3
000162811 7001_ $$aBabatz, Felix$$b4
000162811 7001_ $$aGesell Salazar, Manuela$$b5
000162811 7001_ $$aVölker, Uwe$$b6
000162811 7001_ $$aHallek, Michael$$b7
000162811 7001_ $$aHerrlinger, Ulrich$$b8
000162811 7001_ $$0P:(DE-2719)2481765$$aVorberg, Ina$$b9$$udzne
000162811 7001_ $$aCoch, Christoph$$b10
000162811 7001_ $$00000-0002-5908-8823$$aReiners, Katrin S$$b11
000162811 7001_ $$aHartmann, Gunther$$b12
000162811 770__ $$aCirculating Molecules and Precision Medicine in Cancer
000162811 773__ $$0PERI:(DE-600)2019364-6$$a10.3390/ijms22179211$$gVol. 22, no. 17, p. 9211 -$$n17$$p9211$$tInternational journal of molecular sciences$$v22$$x1422-0067$$y2021
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