Chemistry used in biosensors

• US 9,340,567 B2
• Filed: 11/02/2012
• Issued: 05/17/2016
• Est. Priority Date: 11/04/2011
• Status: Active Grant

• Current Assignee: Ohmx Corporation
• Original Assignee: Ohmx Corporation
• Inventors: Bertin, Paul A., Ahrens, Michael J.
• Primary Examiner(s): Noguerola, Alex
• Application Number:
  US13/667,713
• Publication Number:
  US 20130112572A1
• Time in Patent Office: 1,292 Days
• US Class Current: 1/1
• CPC Class Codes: C07F 17/02 of metals of Groups 8, 9 or 10 of the Periodic System
• CPC Class Codes: C08F 4/61925 two cyclopentadienyl rings being mutually non-bridged
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: G01N 27/3271 Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

The invention relates to novel compositions of 1,3-disubstituted ferrocenes useful for the modification of electrodes.

Claims

1. A ferrocene compound of formula:

2. A ferrocene compound of claim 1, wherein R¹ is hydrogen or —

S—

C₁-C₂₀ alkyl.

3. A ferrocene compound of claim 1, wherein X is —

C₁-C₂₀ alkyl-, —

C₂-C₂₀ alkenyl-, or —

C₂-C₂₀ alkynyl-.

4. A ferrocene compound of claim 3, wherein X is nonylene.

5. A ferrocene compound of claim 1, wherein X is —

X¹—

CONH—

, —

X₁—

CO₂—

, or —

X¹—

OCNH—

, and wherein X¹ is selected from the group consisting of polyoxyalkylene, polymethylene, oligophenylene, and polyphenylene(ethynylene).

6. A ferrocene compound of claim 5, wherein X is —

X¹—

CONH—

, and X¹ is polyoxyalkylene.

7. A ferrocene compound of claim 1, wherein R³ is —

NR⁴R⁵, —

CO₂R⁵, or —

CONR⁴R⁵.

8. A ferrocene compound of claim 7, wherein R³ is —

NH₂, —

CO₂(C₁-C₆ alkyl) or —

CO₂H.

9. A ferrocene compound of claim 1, wherein R³ is —

CONR⁴R⁵;

R⁴ is hydrogen or C₁-C₆ alkyl; and

R⁵ is C₁-C₆ alkyl, aryl(C₁-C₆ alkyl), or heteroaryl(C₁-C₆ alkyl).

10. A ferrocene compound of claim 1, wherein R³ is —

NR⁵CO₂—

R⁶; and

R⁵ is hydrogen, C₁-C₆ alkyl, or aryl(C₁-C₆ alkyl).

11. A ferrocene compound of claim 10, wherein R⁶ is aryl(C₁-C₆ alkyl) or C₂-C₆ alkenyl, where each is optionally substituted with peroxide sensitive moiety.

12. A ferrocene compound of claim 11, wherein the peroxide sensitive moiety is 4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl.

13. A ferrocene compound of claim 1, wherein the peroxide sensitive moiety is selected from the group consisting of:

14. A ferrocene compound of claim 1, wherein R³ is

15. A ferrocene compound of claim 1, which is:

17. The device of claim 16, wherein said 1,3-disubstituted ferrocene comprises a compound of claim 1.

16. A device comprising an electrode comprising:

(i) an optional self-assembled monolayer (SAM); and

(ii) an electroactive active moiety (EAM), said EAM comprising a 1,3-disubstituted ferrocene.

18. The device of claim 16, wherein said 1,3-disubstituted ferrocene further comprises a functional group.

19. The device of claim 18, wherein the functional group comprises a self-immolative moiety and a peroxide sensitive moiety.

20. The device of claim 18, wherein said functional group comprises a capture ligand.

21. The device of claim 18, wherein said functional group is selected from the group consisting of moieties comprising a maleimide, an imidoester, N-hydroxysuccinimidyl, alkyl halide, aryl halide, alpha-haloacyl, and pyridyl disulfide.

22. The device of claim 16, wherein said electrode further comprises a functional group.

23. The device of claim 16, further comprising a self-assembled monolayer (SAM).

24. The device of claim 23, wherein the SAM is a non-conductive oligomer or a conductive oligomer.

25. A device comprising an electrode comprising a self-assembled monolayer (SAM), wherein said SAM comprises a compound having the formula:

Anchor-Spacer 1-EAM-(Spacer₂)n-FG 

 

(I),

wherein said anchor comprises a cyclic-disulfide group,

EAM is a 1,3-disubstituted ferrocene,

FG is a functional group,

Spacer 1 is a SAM forming species,

Spacer 2 is a linker, and

n=0 or 1.

26. A device comprising an electrode comprising a self-assembled monolayer (SAM), wherein said SAM comprises a compound having the formula:

Anchor-Spacer 1-EAM-Spacer 2-FG 

 

(II),

wherein said anchor is linked to said electrode group through a disulfide group,

EAM is a 1,3-disubstituted ferrocene,

FG is a functional group,

Spacer 1 is either insulating or conducting, and

Spacer 2 is an optional linker.

27. A method for detecting one or more target analytes m a test sample, said method comprising:

(a) contacting the test sample with a capture binding ligand under conditions such that the capture binding ligand specifically binds to a target analyst, if present, in said test sample to form a first complex, the capture binding ligand bound to a first solid support;

(b) contacting said first complex, if present, to a soluble capture ligand to form a second complex, wherein said soluble capture ligand comprises a peroxide-generating system;

(c) contacting said second complex with a substrate for said peroxide-generating system under conditions wherein a peroxide is generated to form an assay mixture;

(d) contacting the assay mixture with a second solid support comprising an electrode comprising (i) a self-assembled monolayer (SAM) of an electroactive active moiety (EAM), or (ii) an EAM and optional SAM, wherein said EAM comprises a 1,3-disubstituted ferrocene, a self-immolative moiety, and a peroxide sensitive moiety (PSM) and has a first E⁰, and wherein said peroxide reacts with said PSM to release said SIM from said EAM and result in said EAM having a second E⁰; and

having a second E⁰;

(f) measuring the electrochemical properties of said EAM at the first E⁰ and at the second E⁰; and

(g) detecting said target analyte from said electrochemical properties.

29. The method of claim 27 or 28, further comprising isolating second complex prior to step (c).

30. The method of claim 27 or 28, wherein 1,3-disubstituted ferrocene is a ferrocene compound of formula:

28. A method for detecting one or more target analytes in a test sample, said method comprising:

(a) contacting the test sample with a soluble capture ligand to form a first complex, wherein said soluble capture ligand comprises a peroxide-generating system;

(b) contacting said first complex, if present, with capture binding ligand under conditions such that the capture binding ligand specifically binds to a target analyst to form a second complex, the capture binding ligand bound to a first solid support;

(c) contacting said second complex with a substrate for said peroxide-generating system under conditions wherein a peroxide is generated to form an assay mixture;

(d) contacting the assay mixture with a second solid support comprising an electrode comprising (i) a self-assembled monolayer (SAM) of an electroactive active moiety (EAM) or (ii) an EAM and optional SAM, wherein said EAM comprises a 1,3-disubstituted ferrocene, a self-immolative moiety, and a peroxide sensitive moiety (PSM) and has a first E⁰, and wherein said peroxide reacts with said PSM to release said SIM from said EAM and result in said EAM having a second E⁰; and

having a second E⁰;

(f) measuring the electrochemical properties of said EAM at the first E⁰ and at the second E⁰; and

(g) detecting said target analyte from said electrochemical properties.