EVTOP: extracellular vesicle to phosphoproteomics with microliters of biofluids

We describe here a material and a strategy, extracellular vesicles to phosphoproteins (EVTOP), which achieves the EV isolation, extraction of EV proteins and enrichment of phosphopeptides on the same bifunctionalized magnetic beads using only trace amount of biofluids. The integrated platform allowed us to achieve ultra-sensitive and quantitative EV phosphoproteomics with a few microliters of plasma or 100 μL cerebrospinal fluid, presenting a powerful tool for broad clinical applications.