Extracellular Vesicle Isolation by Size Exclusion Chromatography

This technical report details the methodology for isolating extracellular vesicles (EVs) using size exclusion chromatography (SEC). It outlines the challenges associated with EV isolation due to their heterogeneous nature and the limitations of existing techniques. The report emphasizes the advantages of SEC, including its gentleness and scalability, which make it suitable for achieving high purity and integrity of EVs for various applications. The authors present a workflow utilizing Bio-Gel A-1.5m Resin in conjunction with an NGC Chromatography System, which enhances control over the isolation process. The document includes application data, materials and methods, and discusses the importance of automation in standardizing EV isolation. Key technical considerations are provided to avoid common pitfalls during the process, including sample preparation and column packing. The report concludes with best practices for effective EV isolation and characterization, supported by data from nanoparticle tracking analysis and multiplex western blotting.