Albumin-based non-covalent complexes and methods of use thereof
A non-covalent complex of an albumin molecule and a hydrophobic ligand, compositions containing the same, and methods of use thereof are provided. The present complex may find use in delivering the hydrophobic ligand to microorganisms that have albumin-binding outer surfaces, such as a cell wall.
1. A method for determining a minimum inhibitory concentration (MIC) of an antimicrobial for a microorganism, the method comprising:
a) contacting a plurality of compositions with the antimicrobial, wherein each composition of the plurality of compositions comprises a microorganism comprising a cell wall and wherein each composition is contacted with a different amount of the antimicrobial
b) contacting each composition formed in step a) with an aqueous solution comprising a non-covalent complex of a hydrophobic molecule and a single albumin protein, wherein the hydrophobic molecule functionally associates with the microorganism;
c) detecting one or more properties of the hydrophobic molecule in each composition formed in step b); and
d) determining the minimum inhibitory concentration of the antimicrobial for the microorganism based on the detecting.
2. The method of claim 1 , wherein the microorganism is a pathogenic microorganism.
3. The method of claim 1 , wherein the hydrophobic molecule is a carotenoid.
4. The method of claim 3 , wherein the carotenoid is a carotene.
5. The method of claim 4 , wherein the carotene is lycopene or β-carotene.
6. The method of claim 1 , wherein the antimicrobial is an antibacterial.
7. The method of claim 1 , wherein the antimicrobial is an antifungal.
8. The method of claim 1 , wherein the albumin protein is a human serum albumin protein.
9. The method of claim 1 , wherein the method further comprises forming the aqueous solution comprising the non-covalent complex of hydrophobic molecule and single albumin protein by:
dissolving the hydrophobic molecule in:
i) a first organic solvent comprising a C 3 -C 5 ketone; or
ii) a combination of the first organic solvent and a second organic solvent in a ratio of from about 0.001:1 to about 1000:1 v/v,
to provide a first solution;
combining the first solution with a second solution to provide a third solution, wherein the second solution is an aqueous solution comprising an albumin protein; and
removing the first organic solvent or the combination of the first organic solvent and the second organic solvent from the third solution to provide a fourth solution, wherein the fourth solution is an aqueous solution comprising a non-covalent complex of the hydrophobic molecule and a single albumin protein.
10. The method of claim 9 , wherein the C 3 -C 5 ketone is acetone.
11. The method of claim 9 , wherein the hydrophobic molecule is dissolved in the combination of a first organic solvent and a second organic solvent, and wherein the ratio of the first organic solvent and the second organic solvent is in the range of about 1:1 to about 5:1.
12. The method of claim 11 , wherein the hydrophobic molecule is dissolved in the combination, and wherein the combination comprises the first organic solvent and the second organic solvent in a ratio of about 2:1.
13. The method of claim 9 , wherein the hydrophobic molecule is dissolved in the combination, and wherein the method comprises:
dissolving the hydrophobic molecule in the second organic solvent, to provide a fifth solution; and
combining the fifth solution with the C 3 -C 5 ketone to provide the first solution prior to combining the first solution with the second solution.
14. The method of claim 9 , wherein the removing is performed by evaporation.