Fig. 7: Workflow for the isolation and proteomic characterization of nervous system–derived extracellular vesicle (EV) subpopulations from human serum.

From 1 mL of serum, total EVs (EVT) are extracted using size-exclusion chromatography (SEC) and subsequently incubated with magnetic beads conjugated with antibodies targeting cell-type-specific surface markers: L1CAM (neuronal EVs, nEVs), GLAST (astrocytic EVs, aEVs), MOG (oligodendrocytic EVs, oEVs), and TMEM (microglial/macrophage EVs, m/mEVs). The resulting enriched EV subpopulations are subjected to mass spectrometry-based proteomic analysis, followed by bioinformatic workflows. Illustration created by the authors with BioRender software (www.biorender.com).