Isolation of different extracellular vesicle (EV) subpopulations

A method of processing an extracellular vesicle-containing sample using a device for extracellular vesicle isolation and isolating one or more subpopulations of the extracellular vesicles, the method comprising:

• flowing the extracellular vesicle-containing sample through a flow chamber of the device for extracellular vesicle isolation under an applied fluid pressure, the device having inlets and at least two outlets which are placed in fluid communication with one another via the flow chamber, the device further having one or more filters in the flow chamber between at least one of the inlets and at least one of the at least two outlets, in which the inlets include different inlets including (i) at least one inlet for introducing the extracellular vesicle-containing sample to the flow chamber for flow through the one or more filters and (ii) at least one inlet for providing an evacuating flow to flow at least one of the subpopulations of the extracellular vesicles from a portion of the flow chamber, and wherein the at least one inlet for providing the evacuating flow is positioned between two filters of the one or more filters in the flow chamber such that the evacuating flow isolates the extracellular vesicles in the extracellular vesicle-containing sample having sizes between the two filter filtering sizes associated with the two filters and causes the extracellular vesicles that have been isolated to flow out a corresponding outlet of the at least two outlets;
• during the step of flowing, flowing the extracellular vesicle-containing sample through the one or more filters in the flow chamber to sort the extracellular vesicles of extracellular vesicle-containing sample by size into two or more subpopulations of the extracellular vesicles;
• and
• using the evacuating flow, flowing at least one of the subpopulations of the extracellular vesicles that have been sorted by size after passing through the one or more filters out of a corresponding one of the outlets.