A method of isolating target extracellular vesicles, EVs, from a biofluid comprising a plurality of target EVs and non-target EVs, the method comprising: introducing a plurality of EV capture microparticles to the biofluid to obtain a precursor mixture, wherein the plurality of EV capture microparticles are functionalised, via a plurality of first linkers, with EV-specific binding agents specific to an EV surface marker, such that a plurality of bound microparticle-EV assemblies is formed in the precursor mixture; extracting the bound microparticle-EV assemblies to obtain a cleaned precursor mixture; introducing a plurality of target capture nanoparticles to obtain an assembly mixture, wherein the plurality of target capture nanoparticles are functionalised, via a plurality of second linkers, with target-specific binding agents receptive to surface markers comprised on the target EVs, such that the target capture nanoparticles bind to the plurality of target EVs; cleaving the plurality of first linkers to dissociate at least the plurality of target EVs from the EV capture microparticles, wherein the plurality of second linkers are not cleaved; after the cleaving, extracting the plurality of EV capture microparticles and applying at least one of dielectrophoresis or centrifugation to extract the plurality of target capture nanoparticles, such that the target EVs are separated from the non-target EVs; after separating the plurality of target EVs from the non-target EVs, cleaving the plurality of second linkers to dissociate the plurality of target capture nanoparticles from the plurality of target EVs; and applying at least one of dielectrophoresis or centrifugation to extract the dissociated target capture nanoparticles and isolate a plurality of unbound target EVs. introducing a plurality of EV capture microparticles to the biofluid to obtain a precursor mixture, wherein the plurality of EV capture microparticles are functionalised, via a plurality of first linkers, with EV-specific binding agents specific to an EV surface marker, such that a plurality of bound microparticle-EV assemblies is formed in the precursor mixture; extracting the bound microparticle-EV assemblies to obtain a cleaned precursor mixture; introducing a plurality of target capture nanoparticles to obtain an assembly mixture, wherein the plurality of target capture nanoparticles are functionalised, via a plurality of second linkers, with target-specific binding agents receptive to surface markers comprised on the target EVs, such that the target capture nanoparticles bind to the plurality of target EVs; cleaving the plurality of first linkers to dissociate at least the plurality of target EVs from the EV capture microparticles, wherein the plurality of second linkers are not cleaved; after the cleaving, extracting the plurality of EV capture microparticles and applying at least one of dielectrophoresis or centrifugation to extract the plurality of target capture nanoparticles, such that the target EVs are separated from the non-target EVs; after separating the plurality of target EVs from the non-target EVs, cleaving the plurality of second linkers to dissociate the plurality of target capture nanoparticles from the plurality of target EVs; and applying at least one of dielectrophoresis or centrifugation to extract the dissociated target capture nanoparticles and isolate a plurality of unbound target EVs.