Functionalization of immobilized proteins

• US 20040126831A1
• Filed: 12/31/2002
• Published: 07/01/2004
• Est. Priority Date: 12/31/2002
• Status: Abandoned Application

• Current Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Inventors: Van Antwerp, William P.
• Application Number:
  US10/335,505
• Publication Number:
  US 20040126831A1
• Time in Patent Office: Days
• US Class Current: 435/14
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/485 involving kinase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 1/58 involving urea or urease
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

A method for optimizing the function of a sensor device, wherein a protein, such as an enzyme, is immobilized in a matrix, the matrix is adhered to the device and the protein comprises at least one reactive moiety. The immobilized protein is reacted with a redox agent, wherein the reacting increases the stability of the immobilized protein. The device can be sterilized after the reacting step. A typical device comprises a glucose sensor in which glucose oxidase is embedded in a polymer matrix adhered to the device, the reactive moiety comprises FADH₂, and the redox agent is the reducing agent, sodium borohydride. Also provided is a method of measuring an analyte in a tissue of a subject comprising introducing a sensor device of the invention into the tissue of the subject and detecting the signal generated by the protein. The amount of signal corresponds to the amount of analyte.

Claims

1. A method for optimizing function of a sensor device comprising:

(a) obtaining a sensor device comprising a protein immobilized in a matrix, wherein the matrix is adhered to the device and the protein comprises at least one reactive moiety; and

(b) reacting the immobilized protein with a redox agent;

wherein the reacting increases the stability of the immobilized protein.

2. The method of claim 1, wherein the redox agent is a reducing agent.

3. The method of claim 1, wherein the redox agent comprises sodium borohydride, diborane, Fe²⁺, a reduced metal, a porphyrin system, a ruthenium/porphyrin complex, or FADH₂.

4. The method of claim 1, wherein the redox agent is an oxidizing agent.

5. The method of claim 1, wherein the redox agent is an oxidized metal, Fe³⁺, or a metallo-organic porphyrin.

6. The method of claim 1, wherein the protein is an enzyme.

7. The method of claim 6, wherein the enzyme is glucose oxidase, α

-hydroxy oxidase, lactate oxidase, urease, creatine amidohydrolase, creatine amidinohydrolase, sarcosine oxidase, glutamate dehydrogenase, pyruvate kinase, long chain alcohol oxidase, lactate dehydrogenase, or fructose dehydrogenase.

8. The method of claim 1, wherein the reactive moiety comprises FADH₂.

9. The method of claim 1, wherein the reactive moiety comprises a carbanion or superoxide.

10. The method of claim 1, wherein the matrix comprises a polymer matrix.

11. The method of claim 1, further comprising sterilizing the device after the reacting step.

12. A sensor device comprising an electrode and a polymer matrix adhered to the electrode, wherein the polymer matrix comprises a protein immobilized in the matrix, and wherein the protein comprises a reactive moiety that has been reacted with a redox agent after the protein was immobilized in the matrix.

13. The sensor device of claim 12, wherein the protein is an enzyme.

14. The sensor device of claim 13, wherein the enzyme is glucose oxidase.

15. A method of measuring an analyte in a tissue of a subject, the method comprising introducing a sensor device of claim 12 into the tissue of the subject and detecting the signal generated by the protein, wherein the amount of signal corresponds to the amount of analyte.

16. The method of claim 15, wherein the protein is an enzyme.

17. The method of claim 15, wherein the protein is an enzyme selected from the group consisting of glucose oxidase, α

-hydroxy oxidase, lactate oxidase, urease, creatine amidohydrolase, creatine amidinohydrolase, sarcosine oxidase, glutamate dehydrogenase, pyruvate kinase, long chain alcohol oxidase, lactate dehydrogenase, and fructose dehydrogenase.

18. The method of claim 15, wherein the analyte is glucose and the protein is glucose oxidase.