Value Proposition
· Specific, FRET-based detection: only quantifies successful EV endosomal escape
· Reliable and adaptable: assay works with variety of Evs, target cell types, and fluorescence detection methods
· Genetic modification-free: does not require any genetic engineering of target cells
· Ground-breaking solution: first proposed commercial solution to quanitify effectiveness of EV engineering on cargo delivery
Unmet Need
· Currently, there are no commercially available assays for measuring extracellular vesicle (EV) content delivery to cells. EV content delivery is particularly important in the context of EV therapeutics, where EVs need to deliver their cargo to cells. In EV engineering designed to enhance EV content delivery, assays must be used to gauge the effectiveness of modifications. This goal is often hindered by the lack of available reliable, adaptable, and efficient EV content delivery assays. Therefore, there is a strong need for the development of a quantitative EV content delivery assay as the global EV diagnostics and therapeutics market progresses.
Technology Description
· Researchers at Johns Hopkins have developed a quantitative assay for the measurements of EV content delivery to target cells. In this assay, target cells are loaded with a FRET dye and incubated with Evs engineered to express the enzyme responsible for dye cleavage on the EV inner membrane. Thus, only Evs that successfully undergo endosomal escape (and thus deliver their contents to the cell) result in the cleaved FRET dye signal. Because the assay does not rely on genetic modification of target cells, it can be used with patient-derived cells in addition to cell lines. All preliminary experiments demonstrate high sensitivity with very low background signal, as well as broad applicability.
Stage of Development
· In vitro tests of the efficacy of this technology have been completed.
· The inventors are interested in partnering with life science technology and research tool companies that sell commercial assay kits.
