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Purification Protocols for Extracellular Vesicles

  • Rebecca E. Lane
  • Darren Korbie
  • Matt TrauEmail author
  • Michelle M. Hill
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1660)

Abstract

This chapter provides a description of some of the standard methods used for the isolation of extracellular vesicles (EVs) from a variety of biological fluids, including cell culture media, urine, plasma and serum. The methods presented include ultracentrifugation, ultrafiltration, proprietary polymer-based reagents, size exclusion chromatography, density gradient separation, and immunoaffinity capture. Ultracentrifugation methods use high speed centrifugation to pellet vesicles, whilst polymer-based reagents are added to the sample to facilitate vesicle precipitation using lower speeds. Ultrafiltration involves the concentration of vesicles from a large volume of biological fluid using a centrifugal filter unit. Size exclusion chromatography and density gradient separation are both designed to allow the separation of vesicles from other nonvesicular debris. Immunoaffinity capture methods use antibody-coated beads to selectively isolate vesicles displaying a surface marker of interest. Ultimately, the choice of purification method for an individual experiment is influenced by time, cost, and equipment considerations, as well as the sample requirements for any downstream analyses.

Key words

Extracellular vesicles Exosomes Microvesicles Isolation 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Rebecca E. Lane
    • 1
  • Darren Korbie
    • 1
  • Matt Trau
    • 1
    • 3
    Email author
  • Michelle M. Hill
    • 2
  1. 1.Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and NanotechnologyThe University of QueenslandBrisbaneAustralia
  2. 2.The University of Queensland Diamantina InstituteThe University of QueenslandBrisbaneAustralia
  3. 3.School of Chemistry and Molecular BiosciencesThe University of QueenslandBrisbaneAustralia

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