Visual glucose sensor and methods of use thereof

• US 9,023,661 B2
• Filed: 10/14/2008
• Issued: 05/05/2015
• Est. Priority Date: 10/18/2007
• Status: Active Grant

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Thomas, Joseph, Cash, Michael T.
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US12/250,953
• Publication Number:
  US 20090104714A1
• Time in Patent Office: 2,394 Days
• US Class Current: 436/546
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

A method for determining the presence or amount of one or more ligands or analytes in a sample including contacting the sample with a biosensor having an environmentally-sensitive dye conjugated to a binding member, wherein the biosensor compound exhibits a detectable color change as a result of binding to the ligand or analyte or as a result of a change in concentration of the ligand or analyte in the sample. The presently disclosed biosensors can be used to detect the presence of or amount of physiologically-important metabolites, such as glucose, fatty acids, and lactate, in biological samples.

Claims

1. A method for determining the presence or amount of glucose in a sample, the method comprising:

(a) providing a biosensor compound of Formula III or IV;

2. The method of claim 1, wherein the biosensor compound is:

3. The method of claim 1, wherein the mutated GGBP is selected from the group consisting of W183C, SM4, and Y10C wherein SM4 is mutated glucose/galactose binding protein (GGBP) having the following substitutions:

N39I, G82E, Q83K, N84D, Q175E, Q177H, L178M, W183C, N259E and N260S.

4. The method of claim 2, wherein the mutated GGBP is W183C.

5. The method of claim 2, wherein the mutated GGBP is SM4.

6. The method of claim 1, further comprising continuously:

(a) contacting the mutated GGBP with the sample suspected of containing glucose; and

(b) visually observing the color change.

7. The method of claim 1, wherein the mutated GGBP undergoes a conformation change as a result of a change in glucose concentration of the sample suspected of containing glucose and wherein the method provides a visual change in color as a result of the change in the glucose concentration.

8. The method of claim 1, wherein the biosensor compound of Formula III or IV is immobilized on a solid matrix.

9. The method of claim 8, wherein the solid matrix comprises a chromatographic test strip.

10. The method of claim 1, wherein the color change includes a change from a first absorption wavelength of about 550 nm before the biosensor compound is contacted with the sample suspected of containing glucose to a second absorption wavelength of about 590 nm after the biosensor compound is contacted with the sample suspected of containing glucose.

11. The method of claim 1, wherein the color change includes a change from a first absorption wavelength of about 490 nm before the biosensor compound is contacted with the sample suspected of containing glucose to a second absorption wavelength of about 520 nm after the biosensor compound is contacted with the sample suspected of containing glucose.

12. The method of claim 1, wherein the detecting of the color change comprises comparing an absorption wavelength or an intensity of an absorption wavelength of the biosensor compound with a color wheel.

13. The method of claim 2, wherein the biosensor compound is a compound of Formula (IIIa) and B is SM4, wherein SM4 is mutated glucose/galactose binding protein (GGBP) having the following substitutions:

N39I, G82E, Q83K, N84D, Q175E, Q177H, L178M, W183C, N259E and N260S.