Heparin Affinity Purification of Extracellular VesiclesMonday, 24 March 2014 at 15:00 Add to Calendar ▼2014-03-24 15:00:002014-03-24 16:00:00Europe/LondonHeparin Affinity Purification of Extracellular VesiclesCirculating Biomarkers 2014 in Boston, Massachusetts, USABoston, Massachusetts, USASELECTBIOenquiries@selectbiosciences.com Extracellular vesicles (EVs) are membrane vesicles released by cells. They carry active biomolecules which can be transferred to recipient cells. Isolation and purification of EVs from in vitro conditioned culture media and in vivo biofluids is still a major challenge and the most widely used isolation method still remains ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs due to co-pelleting of proteins and lipids. Affinity purification of biomolecules is an efficient way to achieve high purity without requiring expensive equipment. Previously we have shown that heparin blocks EV uptake in mammalian cells in culture, suggesting a possible direct EV/heparin interaction. Here we show that EVs can be purified from conditioned media using heparin-coated agarose beads. We directly compared heparin-purified EVs to UC prepared and kit-isolated EVs and we show that we can efficiently isolate EVs a higher purity than UC, kit-isolated EVs and even sucrose gradient-purified EVs. Importantly these, heparin-purified EVs retained the RNA content, morphology, and uptake dynamics of UC-isolated EVs. In conclusion, we have discovered a simple and effective way to isolate a highly pure population of EVs using their affinity for heparin. |