Use of hydrogels for biosensors having elevated sensitivity

• US 9,244,064 B2
• Filed: 12/13/2011
• Issued: 01/26/2016
• Est. Priority Date: 12/17/2010
• Status: Active Grant

• Current Assignee: EyeSense AG
• Original Assignee: EyeSense AG
• Inventors: Mller, Achim, Herbrechtsmeier, Peter, Knuth, Monika, Nikolaus, Katharina
• Primary Examiner(s): NOAKES, SUZANNE MARIE
• Application Number:
  US13/994,071
• Publication Number:
  US 20140024060A1
• Time in Patent Office: 1,505 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose

Abstract

The present invention relates to measures for determining glucose and for diagnosing diseases based on impaired glucose metabolism. In particular the present invention relates to the use of a polymer-based hydrogel consisting of at least one water-soluble polymer in a sensor for enriching a dissolved glucose-binding protein, wherein the at least one water-soluble polymer is capable of interacting with the glucose-binding protein and wherein the glucose-binding protein is present in a complex with a ligand.

Claims

1. A method for enriching a dissolved glucose-binding protein in a hydrogel, comprising:

(a) providing a polymer-based hydrogel comprising at least one water-soluble polymer, wherein the polymer-based hydrogel comprises a first hydrogel matrix and a second hydrogel matrix which forms an interpenetrating network within the first hydrogel matrix; and

(b) incorporating a glucose-binding protein and a ligand binding to the glucose-binding protein into the polymer-based hydrogel of step (a),

wherein (i) the at least one water-soluble polymer is capable of interacting with the glucose-binding protein, and (ii) the glucose-binding protein is present in a complex with the ligand,

thereby enriching the dissolved glucose-binding protein in the hydrogel.

2. The method of claim 1, wherein the ligand of the glucose-binding protein is an oligosaccharide, a glycosylated macromolecule, or a glycosylated nanoparticle.

3. The method of claim 1, wherein the dissolved glucose-binding protein is present at an at least 5-fold, at least 10-fold, at least 15-fold, or at least 20-fold elevated concentration as compared to an aqueous solution.

4. The method of claim 1, wherein the at least one polymer is selected from the group consisting of alginates, sepharose, hyaluronic acid, chitosan, and caragenans.

5. The method of claim 1, wherein the polymer-based hydrogel comprises a first hydrogel matrix comprising alginate and a second hydrogel matrix which forms an interpenetrating network within the first hydrogel matrix.

6. The method of claim 1, wherein the glucose-binding protein is selected from the group consisting of lectins, enzymes which bind glucose as substrate, antibodies which specifically recognize glucose, and aptamers which specifically recognize glucose.

7. The method of claim 1, wherein the glucose-binding protein is concanavalin A.

8. The method of claim 7, wherein the concanavalin A concentration is greater than about 0.5 mg/(g polymer).

9. The method of claim 8, wherein the concanavalin A concentration lies between about 2 and about 60 mg/(g polymer).

10. The method of claim 7, wherein the concanavalin A exhibits increased water solubility as compared to unmodified concanavalin A owing to chemical modification.

11. The method of claim 10, wherein the modification is selected from the group consisting of:

pegylation, acetylation, succinylation, and polyoxazolinylation.

12. The method of claim 2, wherein the glucose-binding protein is linked to an oxazine dye and the ligand of the glucose-binding protein is linked to a rhodamine dye.