EVs
in-EV catalogue includes three types of EVs
o Standard EVs (Std-EVs)
o Labelled EVs bioengineered with internal cargoes (Cargo-EVs)
o Bioengineered EVs with fully native membrane proteins (MP-EVs)
Description
Small Extracellular Vesicles or EVs with a size of 30 - 150 nm diameter (including exosomes) are naturally released by most living cells. They are involved in cell physiological and pathological processes, transferring information from donor cells to recipient cells. They carry a variety of cargoes such as proteins, RNAs, lipids and DNA which can be captured by other cells.
The bioengineering of EVs is possible thanks to Ciloa technology, which relies on active sorting of any type of protein to the EVs produced by eukaryotic cells using a proprietary Pilot Peptide (PP, patented). This PP interacts with the ESCRT machinery and sorts the protein of interest to the EVs. A synthetic DNA optimised for coding any protein of interest, is fused to the PP and the resulting chimeric gene is cloned into Ciloa proprietary vector. This vector allows transcription in eukaryotic cells of a mRNA coding for the chimeric protein. Stably transfected HEK293T cells lead to the expression of the target protein into EVs. Proteins can be embedded at the EV surface or can be loaded into the EV lumen. Compared to the engineering technologies often used, the exogenous protein does not interfere with the functions of regular EV proteins (e.g. tetraspanins), it exhibits its fully native conformation without being hindered by a grafting on an EV protein (e.g. CD63), and functional hetero-oligomers of membrane protein complexes (e.g. hetero-octamer of an ion channel) can be sorted on EVs.
Std-EVs
Standard EVs are native EVs, produced by non-modified cells. These EVs are interesting tools that can be used for data normalisation, EV instrument calibration or as a control, either positive or negative, in EV quality controls, EV characterisation, functional assays, etc.
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Cargo-EVs
Labelled EVs bioengineered with internal cargoes are customised using Ciloa technology which does not overexpress and has no interaction with regular EV surface proteins like tetraspanins. These EVs can be used for data normalisation, purification process internal standard, instrument calibration and cell fluorescence imaging.
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MP-EVs
EVs bioengineered with fully native membrane proteins can harbour any kind of protein like G-protein coupled receptors (GPCR), kinase receptors, ion channels or soluble proteins, fused to a transmembrane domain to anchor them in the EV membrane.