Bottom-up assembly of synthetic extracellular vesicles

A synthetic extracellular vesicle having a hydrodynamic radius between 70 nm and 5000 nm, comprising:

• a lipid bilayer comprising at least two lipids selected from the group comprising:
• a neutral lipid selected from the group comprising ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, diacylglycerols, phosphatidylcholines, lysophosphatidylcholines, phosphatidylethanolamines, lysophosphatidylethanolamine, lysoethanolamines, inverted headgroup lipids, sphingosins, sterol-modified phospholipids, ether ester lipids, diether lipids, vinyl ether (plasmalogen);
• an anionic lipid selected from the group comprising phosphatidic acids, lysophosphatidic acid derivatives, phosphatidylglycerols, lysophosphatidylglycerols, phosphatidylserines, lysophosphatidylserines, phosphatidylinositols, phosphatidylinositolphosphates, cardiolipins, Bis(Monoacylglycero)Phosphate derivatives;
• a cationic lipid selected from the group comprising dioleyl-N,N-dimethylammonium chloride;
• N-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride;
• N,N-distearyl-N,N-dimethylammonium bromide;
• N-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride;
• 3β-(N-(N′,N′-dimethylaminoethane)-carbamoyl) cholesterol;
• 1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide;
• 2,3-dioleyloxy-N-[2 (sperminecarboxamido) ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate;
• dioctadecylamidoglycyl carboxyspermine;
• N-(2,3-dioleyloxy)propyl)-N,N-dimethylammonium chloride and 1,2-Dioleoyl-3-dimethylammonium-propane;
• a pH-sensitive lipid selected from the group comprising lipid N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-1-aminium, 1,2-distearoyl-3-dimethylammonium-propane, 1,2-dipalmitoyl-sn-glycero-3-succinate, 1,2-dioleoyl-sn-glycero-3-succinate, N-palmitoyl homocysteine;
• a photoswitchable lipid;
• acylglycine derivatives, prenol derivatives, prostaglandine derivatives, glycosylated diacyl glycerols, eicosanoid derivatives, (palmitoyloxy) octadecanoic acid derivatives, diacetylene derivatives, diphytanoyl derivatives, fluorinated lipids, brominated lipids, lipopolysaccharides;
• one of the aforementioned lipids coupled to a functional ligand selected from the group comprising biotin, N-hydroxysuccinimide ester, nitrilotriacetic acid-nickel, amine, carboxylic acid, maleimides, aromatic maleimid, dithiopyridinyl, pyridyl disulfide, pyridyldithiopropionate, N-benzylguanine, cyanuric chloride, carboxyacyl, cyanur, folate, square, galloyl, glycan, thiol, arginylglycylaspartic acid, a fluorescent dye molecule, a magnetic resonance imaging reagent, a chelator;
• one of the aforementioned lipids coupled to polyethyleneglycol with a molecular weight comprised between 350 and 50,000 g/mole;
• and
• one or more extracellular vesicle associated proteins selected from the group comprising CD9, CD37, CD47, CD53, CD63, CD81, CD82, CD151, Tspan8, heterotrimeric G protein subunit alpha, integrin α-chains, integrin β-chains, transferrin receptor 1, transferrin receptor 2, lysosome associated membrane proteins, heparan sulfate proteoglycans, syndecans, extracellular matrix metalloproteinase inducer, A Disintegrin And Metalloproteinase Domain 10, CD3, CD11c, CD14, CD29, CD31, CD41, CD42a, CD44, CD45, CD50 or intercellular adhesion molecule 1, CD55, CD59, CD73, CD80, CD86, CD90, sonic hedgehog, major histocompatibility complex I, major histocompatibility complex II, epidermal growth factor receptor 2, epithelial cell adhesion molecule, glycophorin A, acetylcholinesterase S and E, amyloid beta precursor protein, multidrug resistance-associated protein 1, stem cells antigen-1, protein complexes endosomal sorting complexes required for transport I, II and III, tumour susceptibility gene 101, charged multivesicular body protein, Apoptosis-Linked Gene 2-Interacting Protein X, vacuolar protein sorting 4 homolog A 4A and 4B, arrestin domain-containing protein, flotillin-1, flotillin-2;
• caveolins, EH-domain containing 1-4, ras homolog family member A, annexins, heat shock proteins, ADP-ribosylation factor 6, syntenin, microtubule-associated protein Tau, cytokines, growth factors, interleukins, milk fat globule-EGF factor 8 protein, adhesion proteins, extracellular matrix proteins, nicotinamide phosphoribosyltransferase, signal transduction proteins, Wnta, Wntb, Fas, Fas Ligand, RANK, RANK Ligand, indolamin-2,3-dioxygenase, cytotoxic T-lymphocyte-associated protein 4-immunoglobulin fusion protein, tumor necrosis factor-related apoptosis-inducing ligand, histone proteins, lamin A/C, inner membrane mitochondrial protein, cytochrome C-1, mitochondrial import receptor subunit TOM20, calnexin, heat shock protein 90 kDa beta, member 1, heat shock 70 kDa protein 5, Golgin A2, Autophagy Related 9A, actinin1, actinin4, cytokeratin 18, or a fragment thereof.