Stabilized lactate responsive enzymes, electrodes and sensors, and methods for making and using the same

• US 9,914,952 B2
• Filed: 06/15/2016
• Issued: 03/13/2018
• Est. Priority Date: 06/15/2015
• Status: Active Grant

• Current Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Inventors: Ouyang, Tianmei, Feldman, Benjamin J., Chen, Kuan-chou
• Primary Examiner(s): FRONDA, CHRISTIAN L
• Application Number:
  US15/183,488
• Publication Number:
  US 20160362716A1
• Time in Patent Office: 636 Days
• Field of Search: None
• CPC Class Codes: C12N 9/0065 acting on hydrogen peroxide as acceptor (1.11)
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: C12Y 111/01006 Catalase (1.11.1.6)
• CPC Class Codes: C12Y 113/12004 Lactate 2-monooxygenase (1.13.12.4)
• CPC Class Codes: G01N 2333/904 acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• CPC Class Codes: G01N 2333/908 acting on hydrogen peroxide as acceptor (1.11)

Abstract

Embodiments of the present disclosure relate to stabilized lactate oxidase compositions, and electrodes, sensors and systems that include the same. Also provided are methods for making the compositions and for detecting and/or measuring lactate in vivo with stable lactate enzyme compositions.

Claims

1. A lactate sensor comprising an enzyme composition that comprises:

a lactate responsive enzyme; and

a stabilizing agent that stabilizes the lactate responsive enzyme in a manner sufficient to provide a sensor signal that changes by 10% or less for 60 continuous hours or more,

wherein the lactate responsive enzyme is lactate oxidase and the stabilizing agent is catalase, present in the enzyme composition in a weight (w/w) ratio of lactate oxidase to catalase ranging from 10 to 1 to 2 to 1.

2. The lactate sensor according to claim 1, wherein the enzyme composition comprises a heterocycle-containing polymer and a crosslinker.

3. The lactate sensor according to claim 2, wherein the lactate responsive enzyme and stabilizing agent are crosslinked within the polymer.

4. The lactate sensor according to claim 2, wherein the lactate responsive enzyme and stabilizing agent are covalently bonded to the polymer.

5. The lactate sensor according to claim 1, wherein the stabilizing agent stabilizes the lactate responsive enzyme in a manner sufficient to provide a sensor signal that changes by 10% or less for 168 continuous hours or more.

6. The lactate sensor according to claim 5, wherein the stabilizing agent stabilizes the lactate responsive enzyme in a manner sufficient to provide a sensor signal that changes by 5% or less for 168 continuous hours or more.

7. The lactate sensor according to claim 1, wherein the stabilizing agent increases sensor sensitivity to lactate as compared to a lactate sensor that does not include the stabilizing agent.

8. The lactate sensor according to claim 7, wherein the stabilizing agent increases sensor sensitivity to lactate for 60 continuous hours or more.

9. The lactate sensor according to claim 8, wherein the stabilizing agent increases sensor sensitivity by 10% or more as compared to a lactate sensor that does not include the stabilizing agent.

10. The lactate sensor according to claim 1, wherein the enzyme composition further comprises an electron transfer agent.

11. The lactate sensor according to claim 10, wherein the electron transfer agent is an osmium-containing complex.

12. The lactate sensor according to claim 10, wherein the electron transfer agent is covalently bonded to the polymer.

13. The lactate sensor according to claim 1, wherein the lactate sensor comprises a working electrode and a counter electrode.

14. The lactate sensor according to claim 13, wherein the enzyme composition is positioned proximate to a working electrode.

15. The lactate sensor according to claim 14, wherein the enzyme composition is fixed to the working electrode.

16. The lactate sensor according to claim 1, wherein the lactate oxidase is present in the enzyme composition in an amount ranging from 0.05 μ

g to 5 μ

g.

17. The lactate sensor according to claim 1, wherein the catalase is present in the enzyme composition in an amount ranging from 0.05 μ

g to 2 μ

g.

18. The lactate sensor according to claim 1, wherein the stabilizing agent stabilizes the lactate responsive enzyme in a manner such that the lactate sensor exhibits a decrease in sensor output of 2 nA or less during exposure to lactate as compared to a lactate sensor that does not include the stabilizing agent.

19. The lactate sensor according to claim 1, wherein the stabilizing agent increases sensor sensitivity to lactate in an amount ranging from 1 nA/mg/dL to 15 nA/mg/dL or more as compared to a lactate sensor that does not include the stabilizing agent.

20. A lactate sensor comprising an enzyme composition that comprises:

a polymer;

a lactate responsive enzyme; and

a stabilizing agent,

wherein the lactate responsive enzyme is lactate oxidase and the stabilizing agent is catalase, present in the enzyme composition in a weight (w/w) ratio of lactate oxidase to catalase ranging from 10 to 1 to 2 to 1, and

wherein the catalase is covalently bonded to the polymer.