Claims
- 1. A method for detecting a carbohydrate in a sample comprising adding a carbohydrate binding protein to said sample, wherein said binding protein changes conformation when bound with said carbohydrate, wherein said binding protein is labeled with an assayable ion that generates a signal upon a conformational change in the proteins; and
detecting the signal from said assayable ion.
- 2. The method according to claim 1, wherein said carbohydrate is glucose or galactose.
- 3. The method according to claim 1, wherein said binding protein is galactose/glucose binding protein, a lectin, a carbohydrate binding domain of a lectin, or a carbohydrate receptor.
- 4. The method according to claim 1, wherein said assayable ion replaces calcium ion in said protein.
- 5. The method according to claim 4, wherein said assayable ion is a lanthanide series ion.
- 6. The method according to claim 5, wherein said lanthanide series ion is a terbium ion.
- 7. The method according to claim 1, wherein said assayable ion generates a fluorescent signal.
- 8. A composition comprising (i) galactose/glucose binding protein bound to a substrate through a cysteine and (ii) a reporter moiety.
- 9. The composition according to claim 8, wherein said reporter moiety is covalently attached to said galactose/glucose binding protein.
- 10. The composition according to claim 8, wherein said reporter moiety is complexed by said galactose/glucose binding protein.
- 11. The composition according to claim 10, wherein the reporter moiety is a lanthanide series ion.
- 12. The composition according to claim 10, wherein said lanthanide series ion is a terbium ion or a europium ion.
- 13. A kit for detecting or quantitating the presence of glucose or galactose in a sample, comprising:
a galactose/glucose binding protein and a lanthanide series ion, wherein said galactose/glucose binding protein is labeled with said lanthanide series ion; and written material describing how to determine the presence of glucose or galactose in said sample using said labeled galactose/glucose binding protein.
- 14. A kit for detecting or quantitating the presence of glucose or galactose in a sample, comprising:
a galactose/glucose binding protein, a lanthanide series ion, a reagent for removal of calcium from said galactose/glucose binding protein, a reagent for inserting a lanthanide series ion into the calcium binding site; and written material describing how to determine the presence of glucose or galactose in said sample using said galactose/glucose binding protein labeled with said lanthanide series ion.
- 15. A method for determining the concentration of glucose or galactose in a sample comprising,
adding galactose/glucose binding protein labeled with a lanthanide series ion to said sample, monitoring the fluorescence of said lanthanide series ion to determine if said galactose/glucose binding protein undergoes a conformation change, correlating the amount of change in fluorescence of said lanthanide series ion with the amount of said glucose or galactose in said sample.
- 16. The method according to claim 15, wherein the change in fluorescence is a change in fluorescence intensity.
- 17. The method according to claim 15, wherein the change in fluorescence is a change in fluorescence lifetime.
- 18. A sensor for determining the presence or concentration of a carbohydrate in sample comprising:
a galactose/glucose binding protein labeled with a reporter moiety, wherein said galactose/glucose binding protein is bound to a solid support; a means for detecting a signal of said reporter moiety generated upon binding of said carbohydrate to the labeled galactose/glucose binding protein; and a means for correlating the signal of the reporter with the amount of carbohydrate present in said sample.
- 19. The sensor of claim 14, wherein said galactose/glucose binding protein (GBP) is modified to include one or more cysteine residues.
- 20. The sensor of claim 14, wherein said galactose/glucose binding protein (GBP) is bound to said solid support by a cysteine residue.
- 21. A method for determining the concentration of glucose or galactose in a sample comprising,
adding to the sample a galactose/glucose binding protein and an organic compound that binds non-covalently to the galactose/glucose binding protein and generates a signal upon a conformational change in the galactose/glucose binding protein; measuring the signal of said organic compound correlating the amount of change in fluorescence of said organic compound with the amount of said glucose or galactose in said sample.
- 22. The method of claim 21 where the organic molecule is anilinonaphthalenesulfonate.
- 23. A method for determining the concentration of glucose or galactose in a sample comprising,
adding to the sample a galactose/glucose binding protein labeled with an electrochemical molecule that generates a signal upon a conformational change in the galactose/glucose binding protein; monitoring the signal of said electrochemical molecule to determine if said galactose/glucose binding protein undergoes a conformation change; correlating the amount of change in signal of said electrochemical molecule with the amount of said glucose or galactose in said sample. signal-generating molecule in its structure.
- 24. A method for determining the concentration of glucose or galactose in a sample comprising,
adding to the sample a signaling fusion protein comprising a galactose/glucose binding protein fused to a signal generating molecule that generates a fluorescence signal upon a conformational change in the galactose/glucose binding protein; measuring the fluorescence signal of said signaling fusion protein; correlating the amount of change in the fluorescence signal with the amount of said glucose or galactose in said sample.
- 25. The method according to claim 24, where the signal generating molecule is Enhanced Green Fluorescence Protein.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 60/330,905 filed Nov. 2, 2001.
GOVERNMENT INTEREST IN THE INVENTION
[0002] This invention was made with Government support under Grant No. NCCW-60 awarded by the National Aeronautics and Space Administration. The Government has certain rights in this invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60330905 |
Nov 2001 |
US |