Chemistry used in biosensors

• US 8,951,400 B2
• Filed: 07/20/2011
• Issued: 02/10/2015
• Est. Priority Date: 10/17/2007
• Status: Active Grant

• Current Assignee: Ohmx Corporation
• Original Assignee: Ohmx Corporation
• Inventors: Ahrens, Michael, Bertin, Paul A., Gray, Harry B., Meade, Thomas J.
• Primary Examiner(s): Shen, Bin
• Application Number:
  US13/187,142
• Publication Number:
  US 20120012472A1
• Time in Patent Office: 1,301 Days
• Field of Search: 205/67
• US Class Current: 205/67
• CPC Class Codes: C07F 15/002 Osmium compounds
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: G01N 27/327 Biochemical electrodes , e.g. electrical or mechanical details for in vitro measurements
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: Y10T 29/49117 Conductor or circuit manufacturing
• CPC Class Codes: Y10T 29/49204 Contact or terminal manufacturing

Abstract

The invention relates to novel compositions of disubstituted bipyridyl osmium complexes useful for the synthesis of labeled proteins, nucleic acids, and for the modification of electrodes.

Claims

1. A compound of chemical formula R¹,R²-bpy-Os(CN)₄:

2. A compound according to claim 1, wherein bpy is chosen from the group consisting of:

3. A compound according to claim 1, wherein said compound is selected from the group consisting of:

4. A composition comprising a compound of claim 1.

5. A composition comprising an electrode comprising a covalently attached electroactive complex (EAM), said EAM comprising a compound of claim 1.

6. The composition according to claim 5, wherein electrode is gold.

7. The composition according to claim 5, wherein the EAM is covalently attached to said electrode via a sulfur atom or two sulfur atoms.

8. The composition according to claim 5, wherein the EAM is attached to said electrode using an attachment linker with a first terminus comprising said sulfur atom(s).

9. The composition according to claim 8, wherein said attachment linker comprises a second terminus that provides a coordination atom for the osmium atom.

10. The composition according to claim 8, wherein said attachment linker is an alkyl chain.

11. The composition according to claim 10, wherein said alkyl chain is substituted.

12. The composition according to claim 5, wherein said electrode comprises a redox active capture complex (REAMC) comprising the EAM and a capture ligand.

13. The composition according to claim 12, wherein said capture ligand provides a coordination atom for the osmium atom.

14. The composition according to claim 12, wherein electrode is gold and said REAMC is covalently attached to said electrode via a sulfur atom or two sulfur atoms.

15. The composition according to claim 14, wherein said REAMC is attached to said electrode using an attachment linker with a first terminus comprising said sulfur atom(s).

16. The composition according to claim 15, wherein said attachment linker comprises a second terminus that provides a coordination atom for the osmium atom.

17. The composition according to claim 15, wherein said attachment linker is an alkyl chain.

18. The composition according to claim 17, wherein said alkyl chain is substituted.

19. The composition according to claim 12, wherein said capture ligand is a peptide or a carbohydrate.

20. The composition according to claim 5, wherein said electrode comprises a first species comprising said EAM and a second species comprising a capture ligand.

21. The composition according to claim 5, Wherein said composition comprises an array of electrodes, each comprising a covalently attached transition metal complex comprising a transition metal and at least one cyano ligand.

22. A composition comprising an electrode comprising:

a) a first species comprising a functional group; and

b) an electroactive complex (EAM), said EAM comprising a compound of claim 1.

23. The composition according to claim 22, wherein said functional group is a maleimide group.

24. A compound having the formula:

Anchor-Spacer1 -EAM-(Spacer2)_n-CL 

 

(I),

wherein said anchor comprises a cyclic-disulfide group,

EAM is compound of claim 1,

CL is a capture ligand,

Spacer 1 is a SAM forming species, and

n=0 or 1.

25. The compound according to claim 24, wherein said cyclic-disulfide group comprises pyridyl-oligophenylethylnyl-[1 ,2,5]-dithiazepane.

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

Anchor-Spacer1-EAM-Spacer2-CL 

 

(II),

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

EAM is a compound of claim 1,

CL is a capture ligand,

Spacer 1 is either insulating or conducting, and

Spacer 2 is optional.

31. A method of detecting a target enzyme comprising:

a) contacting a sample with a composition comprising an electrode comprising;

i) an electroactive moiety (EAM) comprising a compound of any one of claim 1 with a first E⁰;

ii) a capture ligand;

under conditions whereby said target enzyme, if present, alters said capture ligand such that said EAM has a second E⁰; and

b) measuring said second E⁰.

32. The method according to claim 31, wherein said composition comprises a support comprising a plurality of electrodes each comprising:

i) an electroactive moiety (EAM) comprising a compound of any one of claims X-Y with a first E⁰⁴);

ii) a capture ligand.

33. A method of detecting a target enzyme comprising:

a) contacting a sample with a composition comprising a REAMC comprising;

i) an electroactive moiety (EAM) comprising a compound of claim 1 with a first E⁰;

ii) a capture ligand;

under conditions whereby said target enzyme, if present, alters said capture ligand such that said EAM has a second E⁰; and

b) measuring said second E⁰.

34. A method of detecting a target analyte in a test sample, comprising:

c) providing an electrode comprises a compound having the formula;

Anchor-Spacer1-EAM-(Spacer2)_n-CL 

 

(I),

wherein said anchor comprises a cyclic-disulfide group,

EAM is compound of claim 1,

CL is a capture ligand,

Spacer 1 is either insulating or conduction, and

n=0 or 1; and

d) contacting said electrode with said test sample; and

determining the presence of said target analyte by measuring the reorganization energy of said EAM.

35. A method of making a biosensor comprising:

a) providing an electrode comprising;

i) a first species comprising a first functional group; and

ii) an electroactive complex (EAM), said EAM comprising a compound of claim 1;

c) contacting said electrode with a biomolecule comprising a second functional group to form a covalent bond between said first species and said biomolecule.

36. The method according to claim 35, wherein said first functional group is selected from the group consisting of moieties comprising a maleimide, imidoester, Nhydroxysuccinimidyl, alkyl halide, aryl halide, alpha-haloacyl and pryidyl disulfide.

37. The method according to claim 35, wherein said first functional group comprises a maleimide and said biomolecule is a protein comprising a cysteine amino acid.

38. The method according to claim 37, wherein said protein is a peptide.

39. A method, comprising:

a) providing a compound having the formula;

Anchor-Spacer1-EAM-(Spacer2)_n-CL 

 

(I),

wherein said anchor comprises a cyclic-disulfide group,

EAM is a compound of claim 1,

CL is a capture ligand,

Spacer 1 is either insulating or conduction, and n=0 or 1; and

b) contacting said compound to an electrode by opening said cyclic disulfide to form an attachment of said anchor to said electrode.

40. The method according to claim 39, wherein said electrode further comprises a self-assembled monolayer (SAM).

27. A compound of formula R¹,R²-bpy-Os(CN)₄

28. A compound according to claim 27 wherein the electrophile is chosen from the group consisting of activated ester, acyl azide, acyl halide, acyl nitrile, aldehydes, alkyl halide, alkyl halide, alkyl sulfonate, anhydride, aryl halide, aziridine, boronate, carboxylic acid, carbodiimide, diazoalkane, epoxide, haloacetamide, halotriazine, imido ester, isocyanate, isothiocyanate, ketone, maleimide, phosphoramidite, silyl halide, sulfonate ester, and sulfonyl halide and the nucleophile is chosen from the group consisting of amine, aniline, alcohol, phenol, hydrazine, hydroxylamine, carboxylic acid, thiol, and glycol.

29. A compound according to claim 27, wherein R¹ and R² are independently moieties comprising substituted alkyl groups wherein said alkyls are substituted with moieties chosen from the group consisting of carboxylic acid, amine, maleimide, succinimide, N-hydroxysuccinimide, epoxide, and anhydride.

30. A compound according to claim 27, wherein the compound is selected from the group consisting of