Reagents and methods for detecting analytes, and devices comprising reagents for detecting analytes

Claim 2

A reagent for detecting an analyte, comprising: an enzyme selected from the group consisting of a flavoprotein, a quinoprotein, and a combination thereof; and a mediator selected from the group and combinations thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are the same or different, and are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclic, heterocyclic, halo, haloalkyl, carboxy, carboxyalkyl, alkoxycarbonyl, aryloxycarbonyl, aromatic keto, aliphatic keto, alkoxy, aryloxy, nitro, dialkylamino, aminoalkyl, sulfo, dihydroxyboron, and combinations thereof.

Claim 3

The reagent of claim 1 , wherein the mediator is selected from the group consisting of 3-(4′-chloro-phenylimino)-3H-phenothiazine, 3-(4′-diethylamino-phenylimino)-3H-phenothiazine, 3-(4′ethyl-phenylimino)-3H-phenothiazine, 3-(4′-trifluoromethyl-phenylimino)-3H-phenothiazine, 3-(4′-methoxycarbonyl-phenylimino)-3H-phenothiazine, 3-(4′-nitro-phenylimino)-3H-phenothiazine, 3-(4′-methoxy-phenylimino)-3H-phenothiazine, 7-acetyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 7-trifluoromethyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 3-(4′-.omega.-carboxy-n-butyl-phenylimino)-3H-phenothiazine, 3-(4′-aminomethyl-phenylimino)-3H-phenothiazine, 3-(4′-(2″-(5″-(p-aminoph-enyl)-1,3,4-oxadiazoyl)phenylimino)-3H-phenothiazine, 3-(4′-.beta.-aminoethyl-phenylimino)-3H-phenothiazine, 6-(4′-ethylphenyl)amino-3-(4′-ethyl-phenylimino)-3H-phenothiazine, 6-(4′-[2-(2-ethanoloxy)ethoxy]ethoxyphenyl)amino-3-(4′-[2-(2-ethanoloxy)e-thoxy]ethoxy-phenylimino-3H-phenothiazine, 3-(4′-[2-(2-ethanoloxy)ethoxy]e-thoxy-phenylimino-3H-phenothiazine, 3-(4′-phenylimino)-3H-phenothiazinebor-onic acid, (3-(3′,5′-dicarboxy-phenylimino)-3H-phenothiazine, 3-(4′-carboxy-phenylimino)-3H-phenothiazine, 3-(3′,5′-dicarboxy-phenylimino)-3H-phenoxazine, 3-(3′,5′-phenylimino)-3H-phenothiazinedisulfonic acid, 3-(3-phenylimino)-3H-phenothiazinesulfonic acid, and combinations thereof.

Claim 4

The reagent of claim 1 , wherein the mediator comprises

Claim 5

The reagent of claim 1 , wherein the mediator comprises

Claim 6

The reagent of claim 1 , wherein the flavoprotein is selected from the group consisting of FAD-glucose oxidase, flavin-hexose oxidase, FAD-glucose dehydrogenase, [FAD]-lactate oxidase, [FAD]-cholesterol oxidase, [FAD]-alcohol oxidase, [FAD]-d-aminoacid oxidase, [FAD]-choline oxidase, and combinations thereof.

Claim 7

The reagent of claim 1 , wherein the quinoprotein is selected from the group consisting of PQQ-membrane bound glucose dehydrogenase, PQQ-soluble glucose dehydrogenase, [PQQ]-lactate dehydrogenase, [PQQ]-aldehyde dehydrogenase, [PQQ]-methylamine dehydrogenase, [PQQ]-alcohol dehydrogenase, and combinations thereof.

Claim 8

The reagent of claim 1 , wherein the enzyme is selected from the group consisting of FAD-glucose oxidase, PQQ-glucose dehydrogenase, and a combination thereof.

Claim 9

The reagent of claim 7 , wherein the mediator is selected from the group consisting of 3-(4′-chloro-phenylimino)-3H-phenothiazine, 3-(4′-diethylamino-phenylimino)-3H-phenothiazine, 3-(4′ethyl-phenylimino)-3H-phenothiazine, 3-(4′-trifluoromethyl-phenylimino)-3H-phenothiazine, 3-(4′-methoxycarbonyl-phenylimino)-3H-phenothiazine, 3-(4′-nitro-phenylimino)-3H-phenothiazine, 3-(4′-methoxy-phenylimino)-3H-phenothiazine, 7-acetyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 7-trifluoromethyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 3-(4′-.omega.-carboxy-n-butyl-phenylimino)-3H-phenothiazine, 3-(4′-aminomethyl-phenylimino)-3H-phenothiazine, 3-(4′-(2″-(5″-(p-aminoph-enyl)-1,3,4-oxadiazoyl)phenylimino)-3H-phenothiazine, 3-(4′-.beta.-aminoethyl-phenylimino)-3H-phenothiazine, 6-(4′-ethylphenyl)amino-3-(4′-ethyl-phenylimino)-3H-phenothiazine, 6-(4′-[2-(2-ethanoloxy)ethoxy]ethoxyphenyl)amino-3-(4′-[2-(2-ethanoloxy)e-thoxy]ethoxy-phenylimino-3H-phenothiazine, 3-(4′-[2-(2-ethanoloxy)ethoxy]ethoxy-phenylimino-3H-phenothiazine, 3-(4′-phenylimino)-3H-phenothiazineboronic acid, (3-(3′,5′-dicarboxy-phenylimino)-3H-phenothiazine, 3-(4′-carboxy-phenylimino)-3H-phenothiazine, 3-(3′,5′-dicarboxy-phenylimino)-3H-phenoxazine, 3-(3′,5′-phenylimino)-3H-phenothiazinedisulfonic acid, 3-(3-phenylimino)-3H-phenothiazinesulfonic acid, and combinations thereof.

Claim 10

The reagent of claim 7 , wherein the mediator comprises

Claim 11

The reagent of claim 8 wherein the mediator comprises

Claim 12

The reagent of claim 1 , further comprising a polymer selected from the group consisting of carboxymethylcellulose, polyethylene oxide, and combinations thereof.

Claim 13

A reagent for detecting glucose, comprising: PQQ-glucose dehydrogenase in an activity of about 20 Units/μL; a buffer having a concentration between about 0.1 mM and about 100 mM, and a pH between about 4.5 and about 9.5; and a mediator having a structure wherein the mediator has a concentration in the buffer between about 0.1 mM and about 30 mM.

Claim 14

A reagent for detecting glucose, comprising: PQQ-glucose dehydrogenase in an activity of about 20 Units/μL; a buffer having a concentration between about 0.1 mM and about 100 mM, and a pH between about 4.5 and about 9.5; and a mediator having a structure wherein the mediator has a concentration in the buffer between about 0.1 mM and about 30 mM.

Claim 15

The reagent of claim 12 , wherein the buffer is selected from the group consisting of sodium phosphate, potassium phosphate, Hepes, MOPS, TES, Pipes, ACES, BES, Dulbecco's, and combinations thereof.

Claim 16

The reagent of claim 12 , wherein the buffer comprises sodium phosphate.

Claim 17

The reagent of claim 12 , further comprising a polymer selected from the group consisting of carboxymethylcellulose, polyethylene oxide, and combinations thereof.

Claim 18

A reagent for detecting an analyte, comprising: an enzyme selected from the group consisting of PQQ-glucose dehydrogenase, FAD-glucose oxidase, and a combination thereof; and a mediator selected from the group consisting of and combinations thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are the same or different, and are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclic, heterocyclic, halo, haloalkyl, carboxy, carboxyalkyl, alkoxycarbonyl, aryloxycarbonyl, aromatic keto, aliphatic keto, alkoxy, aryloxy, nitro, dialkylamino, aminoalkyl, sulfo, dihydroxyboron, and combinations thereof.

Claim 19

A reagent for detecting an analyte, comprising: an enzyme selected from the group consisting of PQQ-glucose dehydrogenase, FAD-glucose oxidase, and a combination thereof; and a mediator selected from the group consisting of and combinations thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are the same or different, and are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclic, heterocyclic, halo, haloalkyl, carboxy, carboxyalkyl, alkoxycarbonyl, aryloxycarbonyl, aromatic keto, aliphatic keto, alkoxy, aryloxy, nitro, dialkylamino, aminoalkyl, sulfo, dihydroxyboron, and combinations thereof.

Claim 20

The reagent of claim 16 , wherein the mediator comprises

Claim 21

The reagent of claim 16 , wherein the mediator comprises

Claim 22

An electrochemical sensor comprising: a working electrode having a surface; and a second electrode coupled to the working electrode, wherein the surface of the working electrode is coated with a solution of a reagent comprising an enzyme selected from the group consisting of a flavoprotein, a quinoprotein, and a combination thereof; and a mediator selected from and combinations thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are the same or different, and are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclic, heterocyclic, halo, haloalkyl, carboxy, carboxyalkyl, alkoxycarbonyl, aryloxycarbonyl, aromatic keto, aliphatic keto, alkoxy, aryloxy, nitro, dialkylamino, aminoalkyl, sulfo, dihydroxyboron, and combinations thereof.

Claim 23

An electrochemical sensor comprising: a working electrode having a surface; and a second electrode coupled to the working electrode, wherein the surface of the working electrode is coated with a solution of a reagent comprising an enzyme selected from the group consisting of a flavoprotein, a quinoprotein, and a combination thereof; and a mediator selected from and combinations thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are the same or different, and are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cyclic, heterocyclic, halo, haloalkyl, carboxy, carboxyalkyl, alkoxycarbonyl, aryloxycarbonyl, aromatic keto, aliphatic keto, alkoxy, aryloxy, nitro, dialkylamino, aminoalkyl, sulfo, dihydroxyboron, and combinations thereof.

Claim 24

The electrochemical sensor of claim 19 , wherein the working electrode is selected from the group consisting of a carbon electrode, a platinum electrode, a palladium electrode, and a gold electrode.

Claim 25

The electrochemical sensor of claim 19 , wherein the second electrode is selected from the group consisting of a reference electrode and a quasi-reference electrode.

Claim 26

The electrochemical sensor of claim 19 , wherein the second electrode is a silver/silver chloride reference electrode.

Claim 27

The electrochemical sensor of claim 19 , wherein the surface of the working electrode has an area of about 0.00113 cm 2 .

Claim 28

An electrochemical sensor comprising: a working electrode having a surface; and a reference electrode coupled to the working electrode, wherein the surface of the working electrode is coated with a solution of a reagent comprising PQQ-glucose dehydrogenase in an activity of about 1 Units/μL to about 100 Units/μL; a buffer having a concentration of about 100 mM and a pH of about 7.4; and a mediator having a structure wherein the mediator has a concentration in the buffer of about 1 mM to about 100 mM.

Claim 29

An electrochemical sensor comprising: a working electrode having a surface; and a reference electrode coupled to the working electrode, wherein the surface of the working electrode is coated with a solution of a reagent comprising PQQ-glucose dehydrogenase in an activity of about 1 Units/μL to about 100 Units/μL; a buffer having a concentration of about 100 mM and a pH of about 7.4; and a mediator having a structure wherein the mediator has a concentration in the buffer of about 1 mM to about 100 mM.

Claim 30

The electrochemical sensor of claim 24 , wherein the activity of the PQQ-glucose dehydrogenase is about 20 Units/μL, and wherein the concentration of the mediator in the buffer is about 24 mM.

Claim 31

The electrochemical sensor of claim 24 , wherein the activity of the PQQ-glucose dehydrogenase is about 10 Units/μL, and wherein the concentration of the mediator in the buffer is about 12 mM.

Claim 32

The reagent of claim 24 , further comprising a polymer selected from the group consisting of carboxymethylcellulose, polyethylene oxide, and combinations thereof.

Claim 35

The device of claim 28 , wherein the mediator is selected from the group consisting of 3-(4′-chloro-phenylimino)-3H-phenothiazine, 3-(4′-diethylamino-phenylimino)-3H-phenothiazine, 3-(4′ethyl-phenylimino)-3H-phenothiazine, 3-(4′-trifluoromethyl-phenylimino)-3H-phenothiazine, 3-(4′-methoxycarbonyl-phenylimino)-3H-phenothiazine, 3-(4′-nitro-phenylimino)-3H-phenothiazine, 3-(4′-methoxy-phenylimino)-3H-phenothiazine, 7-acetyl-3′-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 7-trifluoromethyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 3-(4′-.omega.-carboxy-n-butyl-phenylimino)-3H-phenothiazine, 3-(4′-aminomethyl-phenylimino)-3H-phenothiazine, 3-(4′-(2″-(5″-(p-aminophenyl)-1,3,4-oxadiazoyl)phenylimino)-3H-phenothiazine, 3-(4′-.beta.-aminoethyl-phenylimino)-3H-phenothiazine, 6-(4′-ethylphenyl)amino-3-(4′-ethyl-phenylimino)-3H-phenothiazine, 6-(4′-[2-(2-ethanoloxy)ethoxy]ethoxyphenyl)amino-3-(4′-[2-(2-ethanoloxy)ethoxy]ethoxy-phenylimino-3H-phenothiazine, 3-(4′-[2-(2-ethanoloxy)ethoxy]ethoxy-phenylimino-3H-phenothiazine, 3-(4′-phenylimino)-3H-phenothiazineboronic acid, (3-(3′,5′-dicarboxy-phenylimino)-3H-phenothiazine, 3-(4′-carboxy-phenylimino)-3H-phenothiazine, 3-(3′,5′-dicarboxy-phenylimino)-3H-phenoxazine, 3-(3′,5′-phenylimino)-3H-phenothiazinedisulfonic acid, 3-(3-phenylimino)-3H-phenothiazinesulfonic acid, and combinations thereof.

Claim 36

The device of claim 28 , wherein the mediator comprises

Claim 37

The device of claim 28 , wherein the mediator comprises

Claim 40

The method of claim 32 , wherein the analyte is glucose.

Claim 41

The method of claim 32 , wherein the mediator is selected from the group consisting of 3-(4′-chloro-phenylimino)-3H-phenothiazine, 3-(4′-diethylamino-phenylimino)-3H-phenothiazine, 3-(4′ethyl-phenylimino)-3H-phenothiazine, 3-(4′-trifluoromethyl-phenylimino)-3H-phenothiazine, 3-(4′-methoxycarbonyl-phenylimino)-3H-phenothiazine, 3-(4′-nitro-phenylimino)-3H-phenothiazine, 3-(4′-methoxy-phenylimino)-3H-phenothiazine, 7-acetyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 7-trifluoromethyl-3-(4′-methoxycarbonylphenylimino)-3H-phenothiazine, 3-(4′-.omega.-carboxy-n-butyl-phenylimino)-3H-phenothiazine, 3-(4′-aminomethyl-phenylimino)-3H-phenothiazine, 3-(4′-(2″-(5″-(p-aminophenyl)-1,3,4-oxadiazoyl)phenylimino)-3H-phenothiazine, 3-(4′-.beta.-aminoethyl-phenylimino)-3H-phenothiazine, 6-(4′-ethylphenyl)amino-3-(4′-ethyl-phenylimino)-3H-phenothiazine, 6-(4′-[2-(2-ethanoloxy)ethoxy]ethoxyphenyl)amino-3-(4′-[2-(2-ethanoloxy)ethoxy] ethoxy-phenylimino-3H-phenothiazine, 3-(4′-[2-(2-ethanoloxy)ethoxy] ethoxy-phenylimino-3H-phenothiazine, 3-(4′-phenylimino)-3H-phenothiazinebor-onic acid, (3-(3′,5′-dicarboxy-phenylimino)-3H-phenothiazine, 3-(4′-carboxy-phenylimino)-3H-phenothiazine, 3-(3′,5′-dicarboxy-phenylimino)-3H-phenoxazine, 3-(3′,5′-phenylimino)-3H-phenothiazinedisulfonic acid, 3-(3-phenylimino)-3H-phenothiazinesulfonic acid, and combinations thereof.

Claim 42

The method of claim 32 , wherein the mediator comprises

Claim 43

The method of claim 32 , wherein the flavoprotein is selected from the group consisting of FAD-glucose oxidase, FAD-glucose dehydrogenase, lactate oxidase, cholesterol oxidase, alcohol oxidase, d-aminoacid oxidase, choline oxidase, and combinations thereof.

Claim 44

The method of claim 32 , wherein the quinoprotein is selected from the group consisting of PQQ-glucose dehydrogenase, lactate dehydrogenase, aldehyde dehydrogenase, methylamine dehydrogenase, alcohol dehydrogenase, and combinations thereof.

Claim 45

The method of claim 32 , wherein the enzyme is selected from the group consisting of FAD-glucose oxidase, PQQ-glucose dehydrogenase, and a combination thereof.

Claim 46

The method of claim 38 , wherein the mediator comprises

Claim 47

The method of claim 38 , wherein the mediator comprises