A size-exclusion-based approach for purifying extracellular vesicles from human plasma

Patrick M. Vanderboom, Surendra Dasari, Gregory N. Ruegsegger, Mark W. Pataky, Fabrice Lucien, Carrie Jo Heppelmann, Ian R. Lanza, K. Sreekumaran Nair

Research output: Contribution to journalArticlepeer-review

Abstract

Extracellular vesicles (EVs) are released into blood from multiple organs and carry molecular cargo that facilitates inter-organ communication and an integrated response to physiological and pathological stimuli. Interrogation of the protein cargo of EVs is currently limited by the absence of optimal and reproducible approaches for purifying plasma EVs that are suitable for downstream proteomic analyses. We describe a size-exclusion chromatography (SEC)-based method to purify EVs from platelet-poor plasma (PPP) for proteomics profiling via high-resolution mass spectrometry (SEC-MS). The SEC-MS method identifies more proteins with higher precision than several conventional EV isolation approaches. We apply the SEC-MS method to identify the unique proteomic signatures of EVs released from platelets, adipocytes, muscle cells, and hepatocytes, with the goal of identifying tissue-specific EV markers. Furthermore, we apply the SEC-MS approach to evaluate the effects of a single bout of exercise on EV proteomic cargo in human plasma.

Original languageEnglish (US)
Article number100055
JournalCell Reports Methods
Volume1
Issue number3
DOIs
StatePublished - Jul 26 2021

Keywords

  • bioinformatics
  • high-intensity exercise
  • mass spectrometry
  • size-exclusion chromatography
  • ultracentrifugation

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Genetics
  • Computer Science Applications
  • Radiology Nuclear Medicine and imaging

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