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This property makes bioengineered EVs promising therapeutic vectors. However, it remains challenging to isolate EVs with a therapeutic payload due to the heterogeneous nature of cargo loading into EVs. In this study, enrichment of EVs with a desired cargo was possible through engineering of the hallmark CD63 transmembrane protein. E\u2010NoMi refers to engineered CD63 with mCherry on the inside of the EV membrane and a tag (3xFLAG) exposed on the outside of the EV membrane. To facilitate EV loading during biogenesis, cargo proteins, such as EGFP, Cre recombinase and the CRISPR\u2010Cas nuclease (SaCas9), were fused to a nanobody (Nb) protein with a high affinity for mCherry. FLAG\u2010tag\u2010based immunocapture from cell conditioned media allowed selection of cargo\u2010loaded E\u2010NoMi\u2010EVs, and tobacco etch virus (TEV) protease cleavage sites were used to remove the 3xFLAG\u2010tag from the surface of E\u2010NoMi\u2010EVs after capture. For functional payload delivery to recipient cells, the vesicular stomatitis virus G (VSV\u2010G) fusogenic protein was incorporated into E\u2010NoMi\u2010EVs to form fusogenic EV\u2010based vectors (EVVs) and proved to be 10\u2010fold more effective at cargo delivery than EVs generated by size\u2010exclusion chromatography. 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