Immunocapture methods to enrich for engineered extracellular vesicles

Abstract

Extracellular vesicles (EVs) are natural liposome-like vesicles secreted by cells into the extracellular space. Provided herein are techniques to enrich cargo-loaded EVs over non-loaded EVs and contaminants. To achieve EVs were engineered to display their surface an antigenic tag for fast and efficient EV isolation by immunocapture and a fluorescent protein in the internal space of the EV. Cargo was loaded into the lumen of the EVs by fusing the cargo with an antibody or nanobody that has an affinity for the fluorescent protein of the internal space. To prevent potential antigenicity of the EVs, a TEV cleavage site was included allowing the removal of exposed antigenic tag from immunocaptured EVs, while preserving their luminal cargo.

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