Characterization of small extracellular vesicles (sEV) from individual mice for the longitudinal investigation of proteome changes in a mouse model of Glioblastoma Multiforme (GBM). We first compared precipitation and size exclusion chromatography (SEC) for sEV purification from 100ul of mouse serum. We then optimized SEC-based procedure for the isolation of sEV’s from 50ul of mouse serum. The optimized procedure was then applied to compare the sEV proteomes for the longitudinal analysis of the GBM-mouse model.