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000302882 1001_ $$0P:(DE-He78)f7694dce6295094b870780d683c93ed2$$aRios de los Rios Resendiz, Jussara$$b0$$eFirst author$$udkfz
000302882 245__ $$aA translational protocol optimizes the isolation of plasma-derived extracellular vesicle proteomics.
000302882 260__ $$a[London]$$bSpringer Nature$$c2025
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000302882 520__ $$aIn translational research and clinical routine, liquid biopsy is a promising tool to direct individually targeted treatments. Among the components of liquid biopsy, extracellular vesicles (EVs) carry manyfold molecular cargo and are increasingly being studied for biomarker identification. In order to identify potential confounding factors and determine optimal conditions when studying blood-derived EV proteins, the impact of pre-analytical variables needs to be assessed. Here we establish an EV enrichment for proteomic analysis workflow in a real-world clinical setting in which we evaluate variables from blood collection through protein preparation and storage for mass spectrometry (MS). We assess hemolysis, particle concentration and size, protein quantity, protein markers and comprehensive proteomic analysis using mass spectrometry to assess the influence of different pre-analytical variables like blood collection tubes, transportation of blood samples and delayed processing. Under these conditions, density gradient and size exclusion chromatography using Sepharose CL-4B show good EV enrichment. For MS, lysis with increased protease inhibitors shows high protein yields while TCA protein precipitation results in high numbers of identified proteins. In summary, we develop here an optimized protocol for the analysis of plasma EV-derived proteomics, evaluating pre-analytical variables relevant for implementation in a clinical setting.
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000302882 650_7 $$2Other$$aExtracellular vesicles
000302882 650_7 $$2Other$$aLiquid biopsy
000302882 650_7 $$2Other$$aPlasma
000302882 650_7 $$2Other$$aPre-analytics
000302882 650_7 $$2Other$$aProteomics
000302882 650_7 $$2Other$$aTranslation
000302882 650_7 $$2NLM Chemicals$$aBlood Proteins
000302882 650_7 $$2NLM Chemicals$$aBiomarkers
000302882 650_7 $$2NLM Chemicals$$aProteome
000302882 650_2 $$2MeSH$$aExtracellular Vesicles: metabolism
000302882 650_2 $$2MeSH$$aExtracellular Vesicles: chemistry
000302882 650_2 $$2MeSH$$aHumans
000302882 650_2 $$2MeSH$$aProteomics: methods
000302882 650_2 $$2MeSH$$aMass Spectrometry
000302882 650_2 $$2MeSH$$aBlood Proteins
000302882 650_2 $$2MeSH$$aTranslational Research, Biomedical: methods
000302882 650_2 $$2MeSH$$aBiomarkers: blood
000302882 650_2 $$2MeSH$$aProteome
000302882 650_2 $$2MeSH$$aLiquid Biopsy: methods
000302882 7001_ $$0P:(DE-He78)f354c88d44ebf813c593e187da2564c9$$aHerrmann-Sim, Freya$$b1
000302882 7001_ $$0P:(DE-He78)a0146d1c0a5c614fe0bb012bbfea56b3$$aWilkesmann, Liliana$$b2$$udkfz
000302882 7001_ $$0P:(DE-He78)daaed5a5b968028e6e95d273150d5ab1$$aHelm, Dominic$$b3$$udkfz
000302882 7001_ $$0P:(DE-He78)0d37cc734b95fed555f2244d6fee6320$$aSchneider, Martin$$b4$$udkfz
000302882 7001_ $$0P:(DE-He78)7a6f726eed43caa75ce1995b0cb09a42$$aCampione, Giorgia$$b5$$udkfz
000302882 7001_ $$0P:(DE-He78)6d6e51fcfc79d92b63429314280d419a$$aPlügge, Klara$$b6$$udkfz
000302882 7001_ $$0P:(DE-He78)d30c5d1ed305fb4a5e00c88e6842de84$$aGreiner, Giovanni$$b7$$udkfz
000302882 7001_ $$0P:(DE-He78)7aad127f8049237b8b21d1f020e476b0$$aLazaro Garcia, Leonie$$b8$$udkfz
000302882 7001_ $$0P:(DE-He78)bdf729ea0d3954f1b296b38db4289789$$aBerker, Julia$$b9$$udkfz
000302882 7001_ $$0P:(DE-He78)027fe772631b4a2d7a45c439cdd75ff2$$aRichter, Karsten$$b10$$udkfz
000302882 7001_ $$0P:(DE-He78)6e1ca2b645d93cb55f1d07262b80b0f0$$aZielske, Lin$$b11
000302882 7001_ $$aHofmann, Wolf-Karsten$$b12
000302882 7001_ $$0P:(DE-He78)97823de5d9dccd50446fc2389ce3ad76$$aClemm von Hohenberg, Katharina$$b13$$eLast author$$udkfz
000302882 773__ $$0PERI:(DE-600)2615211-3$$a10.1038/s41598-025-08366-8$$gVol. 15, no. 1, p. 24292$$n1$$p24292$$tScientific reports$$v15$$x2045-2322$$y2025
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