Isolation of extracellular vesicles by aqueous two-phase system

Abstract

Extracellular vesicles are cell derived membrane vesicles and might be a biomarker for diagnosis especially in tumor detection. However, current methods for isolation of EVs require large starting volume and specialized equipment. To tackle this problem, we developed simple and rapid method by aqueous two-phase system (ATPS). Using PEG and DEX ATPS, we isolate EVs from mixture of EVs and serum. To enhance purity of EVs, batch procedure was combined as additional process to eliminate remaining proteins. We also performed RT-PCR and western blotting to verify sufficient concentration of Melan A and CD81 respectively. Compare to ultra-centrifugation which are the most conventional isolation method, ATPS could isolates EVs from small volume of sample without expensive equipment in short time