Development of a magnetic bead-based method for the collection of circulating extracellular vesicles

Chun Liang Shih*, Kowit Yu Chong, Shih Che Hsu, Hsin Jung Chien, Ching Ting Ma, John Wen Cheng Chang, Chia Jung Yu, Chiuan Chian Chiou

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

39 Scopus citations

Abstract

Cells release different types of extracellular vesicles (EVs). These EVs contain biomolecules, including proteins and nucleic acids, from their parent cells, which can be useful for diagnostic applications. The aim of this study was to develop a convenient procedure to collect circulating EVs with detectable mRNA or other biomolecules. Magnetic beads coated with annexin A5 (ANX-beads), which bound to phosphatidylserine moieties on the surfaces of most EVs, were tested for their ability to capture induced apoptotic bodies in vitro and other phosphatidylserine-presenting vesicles in body fluids. Our results show that up to 60% of induced apoptotic bodies could be captured by the ANX-beads. The vesicles captured from cultured media or plasma contained amplifiable RNA. Suitable blood samples for EV collection included EDTA-plasma and serum but not heparin-plasma. In addition, EVs in plasma were labile to freeze-and-thaw cycles. In rodents xenografted with human cancer cells, tumor-derived mRNA could be detected in EVs captured from serum samples. Active proteins could be detected in EVs captured from ascites but not from plasma. In conclusion, we have developed a magnetic bead-based procedure for the collection of EVs from body fluids and proved that captured EVs contain biomolecules from their parent cells, and therefore have great potential for disease diagnosis.

Original languageEnglish
Pages (from-to)116-122
Number of pages7
JournalNew Biotechnology
Volume33
Issue number1
DOIs
StatePublished - 25 01 2016

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V.

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