Liposome-mediated ligation
Chemoselective ligation of hydrophobic reactants in a lipid phase.
1. A method for making a multicomponent compound comprising:
mixing at least one first hydrophobic reactant comprising a carbohydrate and an N-terminal cysteine residue, at least one second hydrophobic reactant comprising a lipopeptide thioester, and a nonpolar, hydrophobic or amphipathic molecule capable of forming a lipidic structure;
subjecting the mixture to conditions effective to form a lipidic structure in which the first and second reactants are embedded; and
subjecting first and second reactants to conditions effective to allow ligation of the first reactant and the second reactant to yield a multicomponent compound comprising the first and second reactant, wherein the ligation reaction takes place within the lipid phase of the lipidic structure;
wherein neither the first nor the second reactant is a transmembrane protein or membrane-spanning fragment thereof.
2. The method of claim 1 further comprising contacting the multicomponent compound with at least one third hydrophobic reactant within a lipid structure under conditions to allow ligation of the multicomponent compound and the third reactant, to yield a multicomponent compound comprising the first, second and third reactants.
3. The method of claim 2 comprising solubilizing the multicomponent compound and the third reactant within a lipidic structure to facilitate ligation of the multicomponent compound to the third reactant.
4. The method of claim 1 wherein the lipidic structure is selected from the group consisting of a lipid monolayer, lipid bilayer, a liposome, a micelle, a film, an emulsion, a matrix and a gel.
5. The method of claim 1 further comprising contacting the lipidic structure with an initiator compound to catalyze the ligation.
6. The method of claim 1 wherein the ligation is performed in the absence of an initiator compound.
7. The method claim 1 wherein the lipid structure comprises an amphipathic molecule.
8. The method of claim 1 wherein at least one reactant comprises a T-epitope.
9. The method of claim 1 wherein at least one reactant comprises a B-epitope.
10. The method of claim 9 wherein the B-epitope is from a microorganism selected from the group consisting of a virus, a bacterium, a fungus, and a protozoan.
11. The method of claim 10 wherein the microorganism is a human immunodeficiency virus or a hepatitis C virus.
12. The method of claim 9 wherein the B epitope is overexpressed on a cancer cell.
13. The method of claim 1 wherein the first hydrophobic reactant comprises a self-antigen.
14. The method of claim 13 wherein the self-antigen comprises a MUC-1 glycopeptide.
15. The method of claim 1 wherein the first hydrophobic reactant comprises a glycoconjugate selected from the group consisting of a glycosylated protein, a glycosylated peptide, a glycosylated lipid, a glycosylated amino acid, a DNA and an RNA.
16. The method of claim 1 wherein the second hydrophobic reactant comprises a lipopeptide adjuvant.
17. The method of claim 1 wherein the second hydrophobic reactant comprises a Toll-like receptor (TLR) ligand.
18. The method of claim 17 wherein the Toll-like receptor ligand comprises Pam 3 Cys or Pam 3 CysSK n , wherein n=0, 1, 2, 3, 4 or5.
19. The method of claim 1 wherein the second hydrophobic component comprises Pam 3 CysSK 4 .
20. A method for making a multicomponent compound comprising:
mixing at least one first hydrophobic reactant comprising a self-antigen and an N-terminal cysteine residue, at least one second hydrophobic reactant comprising a lipopeptide thioester comprising Pam 3 CysSK n wherein n=0, 1, 2, 3, 4, or 5, and a nonpolar, hydrophobic or amphipathic molecule capable of forming a lipidic structure;
subjecting the mixture to conditions effective to form a lipidic structure in which the first and second reactants are embedded; and
subjecting first and second reactants to conditions effective to allow ligation of the first reactant and the second reactant to yield a multicomponent compound comprising the first and second reactant, wherein the ligation reaction takes place within the lipidic structure;
wherein neither the first nor the second reactant is a transmembrane protein or membrane-spanning fragment thereof.
21. The method of claim 20 wherein at least one reactant comprises at least one B-epitope, at least one T-epitope, or a combination thereof.