Isolation of Biologically Active Exosomes from Plasma of Patients with Cancer

Methods Mol Biol. 2017:1633:257-265. doi: 10.1007/978-1-4939-7142-8_16.

Abstract

A method for exosome isolation from human plasma was developed for rapid, high-throughput processing of plasma specimens obtained from patients with cancer. This method removes the bulk of plasma proteins associated with exosomes and can be used for comparative examinations of exosomes and their content in serial specimens of patients' plasma, allowing for monitoring changes in exosome numbers, profiles, and functions in the course of cancer progression or during therapy. The plasma-derived exosomes can be recovered in quantities sufficient for the characterization of their morphology by transmission electron microscopy (TEM), size and concentration by qNano, protein/lipid ratios, nucleic acid extraction, molecular profiling by Western blots or immune arrays, and functional assays.

Keywords: Cancer; Exosome cargo; Exosome functions; Isolation; Plasma exosomes; Size exclusion chromatography.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers, Tumor / blood*
  • Blotting, Western
  • Cell Fractionation
  • Exosomes / chemistry
  • Exosomes / metabolism*
  • Exosomes / ultrastructure
  • Humans
  • Lipids / blood*
  • Microscopy, Electron, Transmission / methods*
  • Neoplasm Proteins / blood*
  • Neoplasms / blood*
  • Neoplasms / diagnosis
  • Plasma / chemistry
  • Plasma / cytology*

Substances

  • Biomarkers, Tumor
  • Lipids
  • Neoplasm Proteins