Extracellular vesicles (EVs) are nanoscale particles released by cells under both physiological and pathological conditions, that mediate intercellular communication by delivering biomolecules (nucleic acids, proteins, and lipids). In the last decade EVs have attracted the interest of scientific community for their potential as diagnostic and therapeutic agents. Despite this interest, analytical issues posed by clinical use of EVs are still relevant and a real standardization of isolation and quality control procedures is far to be defined. A plethora of analytical techniques have been employed for EV isolation and characterization. The aim of this mini review is to provide an overview of the chromatographic approaches that can be applied to intact EVs. Size-exclusion chromatography, affinity chromatography and ion exchange chromatography are the chromatographic modes that have been successfully employed for EV separation. These chromatographic techniques show strengths and weaknesses that are herein discussed, to build a critical picture of potential future role of chromatographic approaches in EV purification and quality control.