Colloidal particles functionalized homogeneously by biomolecules
The present invention relates to a method for producing a colloid comprising functionalised liquid colloidal particles. The invention also relates to such a colloid and to the uses thereof.
1. A method for obtaining a colloid comprising functionalized liquid colloidal particles comprising the following steps:
a) dispersing an oil in an aqueous solution comprising a fragmentation surfactant, leading to obtaining an emulsion comprising oil droplets suspended in an aqueous phase;
b) dissolving lipids aimed at functionalizing the oil droplets in a polar aprotic solvent, leading to obtaining a functionalization solution;
c) preparing a functionalization mixture comprising the emulsion and the functionalization solution, the volume fraction of the polar aprotic solvent in the functionalization mixture being comprised between 1 and 15%;
d) incubating the functionalization mixture, during which at least a part of the lipids initially present in the functionalization solution are adsorbed on the surface of the oil droplets initially present in the emulsion; and
e) eliminating non-adsorbed lipids during step d);
thus allowing to obtain a colloid comprising functionalized liquid colloidal particles consisting of the oil droplets obtained after step a) on the surface of which the lipids are adsorbed during step d).
2. The method according to claim 1 , wherein the polar aprotic solvent is selected from the group consisting of dimethyl sulfoxide (DMSO), ethyl acetate, acetonitrile, pyridine, butanone, triethylamine, N,N-dimethylformamide (DMF), and mixtures thereof.
3. The method according to claim 1 , wherein the oil dispersed in aqueous solution during step a) is a mineral oil, a vegetable oil, a silicon oil, a halogenated oil, or a mixture thereof.
4. The method according to claim 1 , wherein the lipids comprise at least one fluorophore group and/or at least one biomolecule optionally grafted via a linker.
5. The method according to claim 1 , wherein the lipids are phospholipids optionally grafted with at least one fluorophore group and/or at least one biomolecule, optionally via a linker.
6. The method according to claim 1 , wherein in step c), a dispersion surfactant is added.
7. The method according to claim 1 , wherein in step c), a buffer solution with a pH comprised between 5 and 9 is added.
8. The method according to claim 1 , wherein incubation step d) is performed at a temperature between 15° C. and 40° C. and for a duration between 10 min and 10 h.
9. The method according to claim 1 , wherein the oil dispersed in the aqueous solution comprises Fe 2 O 3 nanoparticles in suspension, thus allowing to obtain a colloid comprising magnetic functionalized liquid colloidal particles.
10. The method according to claim 1 , wherein it comprises, after step e), an additional step f) of grafting biomolecules onto the lipids.
11. The method according to claim 1 , wherein in step c), the volume fraction of the polar aprotic solvent in the functionalization mixture is comprised between 8 and 15%.
12. The method according to claim 1 , wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO).
13. The method according to claim 5 , wherein the phospholipids are phosphatidylethanolamines, optionally grafted with at least one fluorophore group and/or at least one biomolecule, optionally via a linker.
14. The method according to claim 5 , wherein the phospholipids are selected from the group consisting of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine carboxyfluorescein (DOPE-CF), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol biotin (DSPE-PEG-biotin), and mixtures thereof.
15. A colloid comprising functionalized liquid colloidal particles obtainable by the method according to claim 1 .