Proteomic Analysis of Extracellular Vesicles in Neurological Diseases

  • Matías Sáenz-Cuesta
  • Enrique Santamaría
  • Joaquin Fernández-Irigoyen
  • David Otaegui
Part of the Neuromethods book series (NM, volume 127)


Extracellular vesicles (EVs) play a key role in cell-to-cell communication carrying lipids, genetic material, and proteins, producing changes in their target cells. In the last years, EVs have been a hot topic owning to their potential source of biomarkers in many diseases. The search for new biomarkers of central nervous diseases is limited due to the difficulty in accessing to biopsies or complications after obtaining cerebrospinal fluid. In this sense, the study of EVs obtained from blood is proposed as a source of surrogate biomarkers of easy access. One of the most studied components of EVs as biomarkers is their protein cargo. In this chapter we present the main protocols for EV isolation from blood and urine samples, these being based on differential centrifugation and commercial kits for polymeric precipitation. Subsequently, the protocol for a proteomic characterization of plasma-derived EVs is also detailed.

Key words

Extracellular vesicle Isolation protocol Proteomics Mass spectrometry 


  1. 1.
    Théry C, Ostrowski M, Segura E (2009) Membrane vesicles as conveyors of immune responses. Nat Rev Immunol 9:581–593CrossRefPubMedGoogle Scholar
  2. 2.
    Yáñez-Mó M et al (2015) Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles 4:1–60CrossRefGoogle Scholar
  3. 3.
    Sáenz-Cuesta M, Mittelbrunn M, Otaegui D (2015) Editorial: novel clinical applications of extracellular vesicles. Front Immunol 6:1–2Google Scholar
  4. 4.
    Joshi P et al (2014) Microglia convert aggregated amyloid-β into neurotoxic forms through the shedding of microvesicles. Cell Death Differ 21:582–593CrossRefPubMedGoogle Scholar
  5. 5.
    Graner MW et al (2009) Proteomic and immunologic analyses of brain tumor exosomes. FASEB J 23:1541–1557CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Chiasserini D et al (2014) Proteomic analysis of cerebrospinal fluid extracellular vesicles: a comprehensive dataset. J Proteome 106:191–204CrossRefGoogle Scholar
  7. 7.
    Street JM et al (2012) Identification and proteomic profiling of exosomes in human cerebrospinal fluid. J Transl Med 10:5CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    van der Meel R, Krawczyk-Durka M, van Solinge WW, Schiffelers RM (2014) Toward routine detection of extracellular vesicles in clinical samples. Int J Lab Hematol 36:244–253CrossRefPubMedGoogle Scholar
  9. 9.
    Mathivanan S, Fahner CJ, Reid GE, Simpson RJ (2012) ExoCarta 2012: database of exosomal proteins, RNA and lipids. Nucleic Acids Res 40:D1241–D1244CrossRefPubMedGoogle Scholar
  10. 10.
    Witwer KW et al (2013) Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J Extracell Vesicles 2:1–25CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Matías Sáenz-Cuesta
    • 1
    • 2
  • Enrique Santamaría
    • 3
  • Joaquin Fernández-Irigoyen
    • 3
  • David Otaegui
    • 2
  1. 1.Immunology UnitDonostia University HospitalSan SebastiánSpain
  2. 2.Multiple Sclerosis GroupBiodonostia Health Research InstituteDonostia-San SebastiánSpain
  3. 3.Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health DepartmentPublic University of Navarra, Proteored-Institute of Health Carlos III (ISCIII), Navarra Institute for Health Research (IdiSNA)PamplonaSpain

Personalised recommendations