fig6

Figure 6. Comparative and internal enrichment analyses of PROSPR blood-derived EV preparations and contaminant-associated protein contribution. (A) Venn diagram showing the overlap between proteins identified in PROSPR-enriched EVs and EV preparations obtained by dUC-DG. The analysis identified a shared core of 168 proteins, with 285 PROSPR-specific and 574 dUC-DG-specific proteins, highlighting both common EV-associated features and method-dependent proteomic signatures. Comparative benchmarking analyses were performed using the protein categorization approach applied in the reference dataset reported by Morales-Sanfrutos et al.^[16]; (B) Relative distribution of EV-associated proteins and nVEPs in PROSPR-enriched blood EV preparations according to MISEV-based protein classification criteria^[17]. EV-associated proteins accounted for 93.42% of the total proteomic signal, whereas non-EV proteins represented 6.58%, indicating low contaminant contribution in PROSPR-enriched samples. PROSPR: PRotein Organic Solvent PRecipitation; EV: extracellular vesicle; dUC-DG: differential ultracentrifugation followed by density gradient; nVEPs: non-vesicular extracellular proteins; MISEV: Minimal Information for Studies of Extracellular Vesicles.