Rapid Isolation of Extracellular Vesicles from Blood Plasma with Size-Exclusion Chromatography Followed by Mass Spectrometry-Based Proteomic Profiling

Methods Mol Biol. 2017:1660:295-302. doi: 10.1007/978-1-4939-7253-1_24.

Abstract

The presented procedure allows rapid isolation of extracellular vesicles (EVs) from plasma using size-exclusion chromatography (SEC). Additionally, an approach for reducing the lipid and salt content of the EV isolate in preparation for mass spectrometry (MS)-based proteomic analysis is presented. An example setup for proteomic profiling of the processed samples by nanoflow liquid chromatography coupled to tandem mass spectrometry (nLC-MS/MS) is also presented. Approximately 1000 protein groups in blood plasma-derived EVs can be identified and quantitated following this procedure and using the described instrumentation.

Keywords: EVs; Exosomes; Extracellular vesicles; Mass spectrometry; Microparticles; Nano-LC-MS; Proteomic profiling; Proteomics; Sample preparation; Size-exclusion chromatography.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromatography, Gel*
  • Extracellular Vesicles / chemistry*
  • Extracellular Vesicles / metabolism*
  • Humans
  • Liquid-Liquid Extraction
  • Mass Spectrometry*
  • Plasma* / chemistry
  • Proteome
  • Proteomics* / methods
  • Tandem Mass Spectrometry

Substances

  • Proteome