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Multiplexed Phenotyping of Small Extracellular Vesicles Using Protein Microarray (EV Array)

  • Rikke Bæk
  • Malene Møller JørgensenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1545)

Abstract

The Extracellular Vesicle (EV) Array is based on the technology of protein microarray and provides the opportunity to detect and phenotype small EVs from unpurified starting material in a high-throughput manner (Jørgensen et al., J Extracell vesicles 2:1–9, 2013). The technology was established to perform multiplexed phenotyping of EVs in an open platform. This protocol outlines the microarray printing procedure followed by the steps of capture and detection of small extracellular vesicles from plasma/serum or cell culture supernatants. The principles of data treatment and analysis are thoroughly described as well.

Key words

Protein microarray Extracellular vesicles Exosomes Capture antibodies Surface antigens Multiplexed phenotyping 

Abbreviations

dAF488

Dextran coupled with Alexa Fluor 488

sEV

small Extracellular vesicle

IgG

Immunoglobin gamma

ON

Overnight

PBS

Phosphate-buffered saline

RT

Room temperature

Notes

Acknowledgments

The authors would like to thank M.D. Kim Varming and Dr. Evo K.L. Søndergaard for helping establish this protocol.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Clinical ImmunologyAalborg University HospitalAalborgDenmark

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