So i'm extracting and purifying extracellular vesicles from tissues and here i found a problem of coelution of protein contaminants with the EVs. I performed iodixanol DG-UC, SEC, Sucrose DG-UC+SEC but still i observe presence of contaminants in the fraction. though the conc has been reduced from initial stage to now but at present i get around 69µg/ml of protein contamination in my fraction after performing sucrose DG-UC+SEC. Can anyone give suggestion how can i reduce this further? I need to purify it for functional assays