High throughput and rapid isolation of extracellular vesicles and exosomes with purity using size exclusion liquid chromatography

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.

Prediction of DOM removal of low specific UV absorbance surface waters using HPSEC combined with peak fitting

High performance size exclusion chromatography （HPSEC） is used in water quality research primarily to determine the molecular weight distribution of the dissolved organic matter （DOM）, but by applying peak fitting to the chromatogram, this technique can also be used as a tool to model and predict DOM removal. Six low specific UV absorbance （SUVA） source waters were treated using coagulation with alum and both the source and treated water samples were analysed using HPSEC. By comparing the molecular weight profiles of the source and treated waters, it was established that several DOM components were not effectively removed by alum coagulation even after high dosage alum treatment. A peak-fitting technique was applied based on the concept of linking the character （molecular weight profile） of the recalcitrant organics in the treated water with those of the source water. This was then applied to predict DOM treatability by determining the areas of the peaks which were assigned to removable organics from the source water molecular weight profile after peak fitting, and this technique quantified the removable and non-removable organics. The prediction was compared with the actual dissolved organic carbon （DOC） removal determined from jar testing and showed good agreement, with variance between 2% and 10%. This confirmed that this prediction approach, which was originally developed for high SUVA waters, can also be applied successfully to predict DOC removal in low SUVA waters.