Lectin microarray analysis of murine CTL EVs separated by ion exchange method

EVs in culture supernatant can be concentrated with removing exsomeres and 97% free proteins by MWCO 750 kDa UF. DEAE chromatography can be divided into bioactive EXO and other EVs as nucleic acid (DNA) cargo in UF-concentrated EVs. Recently, instead of ultracentrifugation, development of new preparation protocol is demanded for research of reliable bioactivity and drug discovery of extracellular vesicles (EVs). In this study, we propose a novel method for large scale preparation of high-performance extracellular vesicles focusing on membrane negative charge. Murine cytotoxic T lymphocyte (CTL) EVs in supernatant are concentrated more than 20 times at over 97% purity without leaking by 750 kDa MWCO ultrafiltration, replaced with PBS, and subjected to ion exchange DEAE column chromatography.