Comparison of isolation methods using commercially available kits for obtaining extracellular vesicles from cow milk

Extracellular vesicles (EV) are important for delivering biologically active substances to facilitate cell-to-cell communication. Milk-derived EV widely known because of their potential immune enhancement. However, procedures isolating milk-derived have not been fully established. To obtain pure and accurately reveal function, such must be The aim the present study was compare methods using commercially available kits EV. Initially, we investigated remove casein, which is major obstacle in determining purity. We separated whey centrifugation only, acetic acid precipitation, EDTA precipitation. Then, isolated by ultracentrifugation, membrane affinity column, size exclusion chromatography (SEC), polymer-based isolation, or phosphatidylserine-affinity isolation. Using count per milligram protein, a good indicator purity, determined that precipitation best method removing casein. nanoparticle tracking analysis, protein quantity RNA comprehensively compared each isolation its purity yield. found SEC-based qEV column (Izon Science) could collect purer at higher quantities. Thus, combination can effectively isolate high amounts extracellular from bovine milk.