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PXD069821
PXD069821 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Isolation defines identity: Functional consequences of EV purification strategies |
| Description | The biological activity of extracellular vesicles (EVs) is largely defined by their molecular cargo, yet the impact of isolation workflows on EV proteomes and function remains incompletely understood. Here, we compared four isolation strategies for EVs derived from malignant ascites and ES-2 ovarian cancer cell culture supernatants, assessing yield, particle size, protein cargo, and EV-associated enzymatic activity. Proteomic analyses of particle-normalized preparations were performed according to MISEV2023 guidelines, and vesicle-associated protease activity was profiled using a FRET-based assay with inhibitor panels. Principal component and overlap analyses identified a common EV proteome signature for ascites and ES-2 EVs, which was complemented by workflow-dependent detection of additional proteins. Ultracentrifugation/density gradient (UC-DG) and tangential flow filtration/size exclusion chromatography (TFF-SEC) achieved the highest enrichment of canonical EV markers, whereas TFF/ultrafiltration (TFF-UF) was enriched in lipoproteins and secreted proteins. Functionally, UC-DG and TFF-SEC samples exhibited strong ADAM10-associated activity, while TFF-UF retained residual non-metalloprotease activity. Together, these findings demonstrate that even under standardized particle input, EV isolation workflows critically shape both EV proteomic profiles and functional activities. Our study provides a systematic, particle-normalized functional comparison of EV isolation methods, emphasizing the importance of workflow choice for biomarker discovery and mechanistic studies. |
| HostingRepository | MassIVE |
| AnnounceDate | 2026-02-02 |
| AnnouncementXML | Submission_2026-02-02_00:14:46.769.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Witold Szymanski |
| SpeciesList | scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606; |
| ModificationList | S-carboxamidomethyl-L-cysteine; Oxidation; Acetyl |
| Instrument | timsTOF Ultra |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2025-10-23 02:30:59 | ID requested | |
| ⏵ 1 | 2026-02-02 00:14:47 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: Extracellular vesicles, Proteome, ADAM10 activity, Standardization, MISEV2023, timstof, dia, proteomics, DatasetType:Proteomics |
Contact List
| Elke Pogge von Strandmann | |
|---|---|
| contact affiliation | Institute for Tumor Immunology, Philipps-University Marburg |
| contact email | poggevon@staff.uni-marburg.de |
| lab head | |
| Witold Szymanski | |
| contact affiliation | Philipps-University Marburg Biochemical/Pharmacological Center Department of Medicine |
| contact email | witold.szymanski@uni-marburg.de |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
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