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Purification Protocols for Extracellular Vesicles.

Rebecca E Lane1, Darren Korbie1, Matt Trau2,3

  • 1Centre 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.

Methods in Molecular Biology (Clifton, N.J.)
|August 23, 2017
PubMed
Summary
This summary is machine-generated.

This chapter details standard methods for isolating extracellular vesicles (EVs) from biological fluids. Key techniques include ultracentrifugation, ultrafiltration, and chromatography, chosen based on experimental needs.

Keywords:
ExosomesExtracellular vesiclesIsolationMicrovesicles

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biochemistry

Background:

  • Extracellular vesicles (EVs) are crucial biomarkers and therapeutic agents.
  • Efficient isolation of EVs from biological fluids is essential for their study and application.

Purpose of the Study:

  • To describe standard methods for extracellular vesicle (EV) isolation from various biological sources.
  • To provide an overview of techniques applicable to cell culture media, urine, plasma, and serum.

Main Methods:

  • Ultracentrifugation: Pelleting EVs via high-speed centrifugation.
  • Polymer-based precipitation: Using reagents to precipitate EVs for lower-speed centrifugation.
  • Ultrafiltration: Concentrating EVs using centrifugal filter units.
  • Chromatographic methods: Size exclusion chromatography and density gradient separation for EV purification.
  • Immunoaffinity capture: Utilizing antibody-coated beads for selective EV isolation based on surface markers.

Main Results:

  • Multiple standard methods exist for EV isolation, each with specific advantages.
  • Technique selection is influenced by factors such as time, cost, equipment availability, and downstream application requirements.

Conclusions:

  • The choice of EV isolation method requires careful consideration of experimental constraints and goals.
  • Standardized and optimized EV isolation protocols are critical for reproducible research and clinical applications.