GLUCOSE SENSING ASSAY

• US 20140350370A1
• Filed: 04/08/2014
• Published: 11/27/2014
• Est. Priority Date: 04/08/2013
• Status: Abandoned Application

• Current Assignee: Texas A&M University System
• Original Assignee: Texas A&M University System
• Inventors: Cummins, Brian M., Cot, Gerard L.
• Application Number:
  US14/248,043
• Publication Number:
  US 20140350370A1
• Time in Patent Office: Days
• US Class Current: 600/365
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/14735 comprising an immobilised reagent
• CPC Class Codes: A61B 5/686 Permanently implanted devices, e.g. pacemakers, other stimulators, biochips A61B5/6861 takes precedence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose

Abstract

The disclosure provides a ligand that competes with glucose for binding the protein Concanavalin A (ConA) and competitive binding assays incorporating the ligand. The competing ligand binds to the primary and part or all of the extended binding sites of Concanavalin A. These and other aspects of the disclosure are useful for glucose monitoring (e.g., continuous glucose monitoring (CGM)).

Claims

1. A ligand for Concanavalin A, comprising:

(a) a Concanavalin A binding component that binds to the primary glucose binding site and part or all of an extended binding site on Concanavalin A; and

(b) a transduction component, wherein the binding component is coupled to the transduction component, and wherein the transduction component generates a detectable signal upon binding of the ligand to Concanavalin A.

2. The ligand of claim 1, wherein the binding component comprises one or more mannose moieties.

3. The ligand of claim 1, wherein the binding component comprises a trimannose moiety.

4. The ligand of claim 3, wherein the trimannose moiety is 3,6-Di-O-(α

-D-mannopyranosyl)-D-mannopyranose.

5. The ligand of claim 1, wherein the binding component comprises a bimannose moiety.

6. The ligand of claim 5, wherein the bimannose moiety is 6-O-α

-D-mannopyranosyl-D-mannopyranose or 3-O-α

-D-mannopyranosyl-D-mannopyranose.

7. The ligand of claim 1, wherein the ligand has a binding affinity for Concanavalin A from about 10,000 to about 10,000,000 L/mol.

8. The ligand of claim 1, wherein the transduction component is a fluorophore, a Raman reporter, or a nanoparticle, or is electrochemically active.

9. The ligand of claim 1, wherein the transduction component generates a detectable signal by a transduction mechanism selected from fluorescence intensity, fluorescent resonance energy transfer (FRET), fluorescence anisotropy, fluorescence lifetime, Raman spectroscopy, and metal enhanced plasmonics.

10. The ligand of claim 1, further comprising a tether point for immobilization of the ligand to a structure or surface.

11. The ligand of claim 1, further comprising a proteinaceous scaffold.

12. The ligand of claim 11, wherein the binding component or the transduction component is coupled to the proteinaceous scaffold, or both the binding component and the transduction component are independently coupled to the proteinaceous scaffold.

13. The ligand of claim 11, wherein the proteinaceous scaffold has a net negative charge.

14. The ligand of claim 11, wherein the proteinaceous scaffold is or comprises ovalbumin, RNase B, or any derivative thereof.

23. The method of claim 15, wherein the ligand is the ligand of claim 1.

24. The method of claim 23, wherein each of the ligand and the Concanavalin A comprises a transduction component.

25. The method of claim 24, wherein the transduction component of the ligand and the transduction component of the Concanavalin A are capable of mutually interacting as a FRET pair.

28. The system of claim 26, wherein the ligand is the ligand of claim 1.

15. A method for monitoring glucose in a sample, comprising:

detecting the competitive binding of a ligand to Concanavalin A in the sample, wherein the ligand has an affinity toward the primary glucose binding site and at least a portion of an extended binding site of Concanavalin A, wherein the ligand competes with glucose for binding to the primary binding site of Concanavalin A, and wherein a detectable signal is provided by a transduction component upon binding of the ligand to Concanavalin A.

16. The method of claim 15, wherein the detecting step comprises contacting the sample with the Concanavalin A and the ligand.

17. The method of claim 15, wherein the equilibrium binding of the ligand to Concanavalin A is inversely related to the glucose concentration in the sample.

18. The method of claim 15, wherein the sample is an in vitro or in vivo biological sample.

19. The method of claim 18, wherein the biological sample is blood, blood plasma, blood serum, extracellular fluid, interstitial fluid, or aqueous humor fluid.

20. The method of claim 15, wherein the detecting is performed in a continuous glucose monitoring assay.

21. The method of claim 15, wherein the ligand comprises the transduction component.

22. The method of claim 15, wherein the Concanavalin A comprises the transduction component.

26. A glucose monitoring system, comprising:

(a) Concanavalin A;

(b) a ligand having an affinity toward the primary binding site and all or part of the extended binding site of Concanavalin A, wherein the ligand effectively competes with glucose for binding to Concanavalin A; and

(c) a transduction component to signal the state of assay binding.

27. The system of claim 26, wherein the ligand comprises the transduction component.

29. The system of claim 26, wherein each of the ligand and the Concanavalin A comprises a transduction component.

30. The system of claim 26, wherein the system is adapted to an implanted biosensor.

31. The system of claim 26, wherein the system is adapted to a subcutaneous biosensor.