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Purification Protocols for Extracellular Vesicles

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Extracellular Vesicles

Part of the book series: Methods in Molecular Biology ((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.

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Author information

Authors and Affiliations

  1. Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Building 75, Cnr Cooper and College Road, St. Lucia, Brisbane, QLD, 4072, Australia

    Rebecca E. Lane, Darren Korbie & Matt Trau

  2. The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia

    Michelle M. Hill

  3. School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia

    Matt Trau

Authors
  1. Rebecca E. Lane
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  2. Darren Korbie
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  3. Matt Trau
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  4. Michelle M. Hill
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Corresponding author

Correspondence to Matt Trau .

Editor information

Editors and Affiliations

  1. CloudHealth Genomics, Ltd., Shanghai, China

    Winston Patrick Kuo

  2. Predicine, Inc., Hayward, California, USA

    Shidong Jia

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Lane, R.E., Korbie, D., Trau, M., Hill, M.M. (2017). Purification Protocols for Extracellular Vesicles. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_10

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  • DOI: https://doi.org/10.1007/978-1-4939-7253-1_10

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7251-7

  • Online ISBN: 978-1-4939-7253-1

  • eBook Packages: Springer Protocols

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