Ciloa’s unique technology makes possible to produce bioengineered small EVs (sEVs) with any type of protein on the surface of EVs or in the their lumen.

It is based on the discovery of a peptide, called pilot peptide that contains motifs that interact with so-called ESCRT proteins. The fusion of the pilot peptide with a protein triggers its active sorting and secretion with sEVs.

By this way, it is possible to target proteins at the EV surface or to load proteins into the EV lumen.

Membrane targeting

Membrane proteins like GPCRs, kinase receptors, ion channels, viral proteins, etc. can be targeted on EVs with their fully native conformation by fusing them to the pilot peptide.

Non-membrane proteins can also be anchored at the EV membrane and displayed at the surface, by adding a transmembrane domain in addition to the pilot peptide.

Lumen loading

To load proteins into the lumen of EVs, the sequence encoding the protein of interest is fused at the N-terminus to a Src peptide (for membrane anchoring) and at the C-terminus to the pilot peptide (for sorting into EVs). 

Thanks to our technology, we do not fuse the cargo protein with natural EV proteins, like tetraspanins. Therefore, there is no modifications of the EV surface with overexpression of EV protein markers and so minimal modifications of the EV natural properties.