Extracellular vesicles are small membrane-enclosed structures that are released from practically all cells. They contain different types of molecules, including small RNAs, and are involved in many physiological processes, which has led to increasing research into their role as potential biomarkers that could help doctors tailor treatment strategies to individual patients and help them identify those who are more likely to respond to a particular treatment approach. However, standardisation and further characterisation of the separation techniques used in extracellular vesicle studies is still ongoing. To this end, we have tested some of the current molecular biological methods for the isolation of extracellular vesicles from cerebrospinal fluid and plasma, following the MISEV guidelines. Fast protein liquid chromatography Ultracentrifugation was found to be a suitable method for isolation of extracellular vesicles from cerebrospinal fluid, and fast protein liquid chromatography in conjunction with ultracentrifugation and sucrose gradient centrifugation were considered to be suitable methods for isolation of extracellular vesicles from plasma, but given the results, further optimisation will be needed.