Abstract
Background: Extracellular vesicles (EVs) are membrane encapsulated nanoparticles that function as carriers and play a role in intercellular communication. There are a large number of EVs in the blood and serve as an indicator of pathophysiological conditions. Studies on the basics and application of EVs are hampered by the limitations of current protocols to isolate EVs from blood. However, current isolation methods are difficult to achieve a balance between yield and purity.Results: Firstly, we use Sepharose-4B to build a self-made size exclusion chromatography (SEC) column and perform separation and identification. Then we use the SEC column to systematically compare the efficiency with the most common EV isolation methods: ultracentrifugation (UC) and total exosomes isolation commercial kit (TEI). The EVs isolated through different methods were characterized the yield and size of EVs, analyzed their protein profiles, the morphology and purity were observed under the transmission electron microscope. To further improve the quality and purity, we combined SEC and UC methods and established a two-steps method to isolated EVs from serum.Conclusion: Our study presents the combination of size-exclusion chromatography and ultracentrifugation as a feasible and time-saving method to isolate high quality and purity extracellular vesicles from serum.
An isolation system to collect high quality and purify extracellular vesicles from serum
