Heparin affinity purification of extracellular vesicles

Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA, and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely used isolation method is ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs.

Previously researchers from Harvard Medical School have shown that heparin blocks EV uptake in cells, supporting a direct EV-heparin interaction. Here they show that EVs can be purified from cell culture media and human plasma using ultrafiltration (UF) followed by heparin-affinity beads. UF/heparin-purified EVs from cell culture displayed the EV marker Alix, contained a diverse RNA profile, had lower levels of protein contamination, and were functional at binding to and uptake into cells. RNA yield was similar for EVs isolated by UC. The researchers were able to detect mRNAs in plasma samples with comparable levels to UC samples.

exosome rnaExtracellular vesicles are efficiently isolated and purified using heparin-coated agarose beads.

Balaj L, Atai NA, Chen W, Mu D, Tannous BA, Breakefield XO, Skog J, Maguire CA. (2015) Heparin affinity purification of extracellular vesicles. Sci Rep 5:10266. [article]

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