Isolation of Proteins from Extracellular Vesicles (EVs) for Mass Spectrometry-Based Proteomic Analyses

Prabal Subedi, Michael Schneider, Michael J. Atkinson, Soile Tapio

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

Extracellular vesicles (EVs) are freely circulating nano/micrometer-sized membrane-bound vesicles released by various cell types. Their cargo consists of proteins, lipids, metabolites, and different types of RNA molecules reflecting the origin of the secreting cell type or tissue. Since the EV cargo is constantly changing in response to pathological status or different environmental stressors, it has been extensively studied in the quest for biomarkers, especially in the cancer research. Mass spectrometry (MS)-based proteome analysis is a powerful tool to elucidate the protein cargo in EVs. This chapter describes and characterizes three MS-compatible lysis methods, namely by using urea, guanidium hydrochloride, and radioimmunoprecipitation buffer for isolating proteins from EVs.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages207-212
Number of pages6
DOIs
StatePublished - 2021
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2261
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Data-dependent acquisition
  • Exosomes
  • Extracellular vesicles (EVs)
  • Mass spectrometry
  • Proteomics
  • Sample preparation

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