High throughput and rapid isolation of extracellular vesicles and exosomes with purity using size exclusion liquid chromatography
Kshipra Kapoor, Kristen Harris, Kent A. Arian, Lihua Ma, Beatriz Schueng Zancanela, Kaira A. Church, Kathleen M. McAndrews, Raghu Kalluri
Abstract
Extracellular vesicles (EVs) have emerged as potential biomarkers for diagnosing a range of diseases without invasive procedures. Extracellular vesicles also offer advantages compared to synthetic vesicles for delivery of various drugs; however, limitations in segregating EVs from other particles and soluble proteins have led to inconsistent EV retrieval rates with low levels of purity. Here, we report a new high-yield (88.47 %) and rapid (<20 min) EV isolation method termed size exclusion – fast protein liquid chromatography (SE-FPLC). We show SE-FPLC can effectively isolate EVs from multiple sources including EVs derived from human and mouse cells and serum samples. The results indicate that SE-FPLC can successfully remove highly abundant protein contaminants such as albumin and lipoprotein complexes, which can represent a major hurdle in large scale isolation of EVs. The high-yield nature of SE-FPLC allows for easy industrial scaling up of EV production for various clinical utilities. SE-FPLC also enables analysis of small volumes of blood for use in point-of-care diagnostics in the clinic. Collectively, SE-FPLC offers many advantages over current EV isolation methods and offers rapid clinical translation. • Size exclusion – fast performance liquid chromatography (SE-FPLC) enables rapid isolation of extracellular vesicles (EVs). • SE-FPLC removes albumin and lipoprotein contaminants from EVs. • High-yield EV isolation by SE-FPLC enables scale-up of EV production.