Purification and Phosphoproteomic Analysis of Plasma-Derived Extracellular Vesicles

Methods Mol Biol. 2022:2504:147-156. doi: 10.1007/978-1-0716-2341-1_11.


A successful phosphoproteomics analysis of extracellular vesicles (EVs) requires a unique approach, fine-tuned to address the challenges that have plagued plasma-based biomarker discovery. Here, I detail a procedure, which combines EVtrap-based high-recovery EV isolation, phase-transfer surfactant method for protein extraction, and PolyMAC-based enrichment of phosphopeptides. The combination of these methods provides a highly effective strategy for EV-based phosphoproteome analysis and leads to the discovery of novel phospho-markers previously undetectable.

Keywords: EVtrap; Exosomes; Extracellular vesicles (EVs); Liquid biopsy; Phosphoproteomics; Plasma.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers / metabolism
  • Extracellular Vesicles* / metabolism
  • Phosphopeptides / metabolism
  • Plasma / chemistry
  • Proteome / metabolism


  • Biomarkers
  • Phosphopeptides
  • Proteome