Highly sensitive amperometric bi-mediator-based glucose biosensor
First Claim
1. An amperometric sensor for the determination of glucose in aqueous media comprising:
- a sensing electrode, the sensing electrode comprising;
a non-conductive support member, with the non-conductive support member comprising;
a non-conductive polymeric film;
the non-conductive polymeric film coated with an electrically conductive layer, with the electrically conductive layer containing;
a first redox mediator; and
a reference electrode comprising;
a non-conductive polymeric film, the polymeric film coated with an electrically conductive formation comprising Ag/AgCl dispersed in a resin formulation, and with the reference electrode having an opening; and
a reagent strip, the reagent strip comprising;
a carrier strip that is a porous or fibrous water absorbent matrix, the carrier strip impregnated with a mixture comprising;
glucose oxidase,horseradish peroxidase,a second redox mediator that can be oxidized by hydrogen peroxide under catalysis by horseradish peroxidase,at least one surfactant,at least one stabilizer,a buffering agent to maintain a pH from about 4 to about 8;
with the electrically conducting layer of the sensing electrode and the electrically conductive formulation of the reference electrode facing each other; and
with the reagent strip superimposed on and in physical contact with the electrically conducting layer of the sensing electrode, and with the reference electrode superimposed on the reagent strip so that the electrically conductive formulation coating the reference electrode is superimposed on the reagent strip and in physical contact with the reagent strip;
so that the sensing electrode and the reagent strip and the reference electrode form a sandwich.
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Accused Products
Abstract
A highly sensitive sensor for the amperometric assay of glucose in aqueous media having a face-to-face sandwich configuration is provided which comprises a sensing electrode strip containing a first redox mediator in an electrically conductive coating and a reference electrode strip in simultaneous contact with a reagent strip containing a second redox mediator which is sandwiched between the active electrode surfaces. An opening is provided in the reference electrode for the introduction of samples. In one embodiment, a whole blood separation membrane is interposed between the reference electrode and the reagent strip to filter red blood cells and other particles from whole blood to enable direct assay of glucose without sample preparation. The sensing electrode comprises a non-conductive support member having an electrically conductive layer containing the first redox mediator. The reference electrode is typically a Ag/AgCl electrode formed by coating an ink containing Ag/AgCl dispersed in a resin on a non-conductive base. The reagent strip is a porous or fibrous carrier, typically a paper, impregnated with a mixture containing the second redox mediator, glucose oxidase, horseradish peroxidase, at least one surfactant and at least one stabilizer comprising an aqueous thickening agent. In one version of the sensor, the sensing electrode comprises a support member of polyester film coated with an electrically conductive graphite composition containing dimethylferrocene as the first redox mediator and the second redox mediator comprises potassium ferrocyanide.
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Citations
22 Claims
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1. An amperometric sensor for the determination of glucose in aqueous media comprising:
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a sensing electrode, the sensing electrode comprising; a non-conductive support member, with the non-conductive support member comprising; a non-conductive polymeric film; the non-conductive polymeric film coated with an electrically conductive layer, with the electrically conductive layer containing; a first redox mediator; and a reference electrode comprising; a non-conductive polymeric film, the polymeric film coated with an electrically conductive formation comprising Ag/AgCl dispersed in a resin formulation, and with the reference electrode having an opening; and a reagent strip, the reagent strip comprising; a carrier strip that is a porous or fibrous water absorbent matrix, the carrier strip impregnated with a mixture comprising; glucose oxidase, horseradish peroxidase, a second redox mediator that can be oxidized by hydrogen peroxide under catalysis by horseradish peroxidase, at least one surfactant, at least one stabilizer, a buffering agent to maintain a pH from about 4 to about 8; with the electrically conducting layer of the sensing electrode and the electrically conductive formulation of the reference electrode facing each other; and
with the reagent strip superimposed on and in physical contact with the electrically conducting layer of the sensing electrode, and with the reference electrode superimposed on the reagent strip so that the electrically conductive formulation coating the reference electrode is superimposed on the reagent strip and in physical contact with the reagent strip;
so that the sensing electrode and the reagent strip and the reference electrode form a sandwich. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for assaying a sample of whole blood for the concentration of glucose contained in the whole blood and fractions thereof, comprising the steps of:
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a) providing an amperometric biosensor for glucose comprising; a sensing electrode, the sensing electrode comprising; a non-conductive support member, with the non-conductive support member comprising; a non-conductive polymeric film; the non-conductive polymeric film coated with an electrically electrode layer, with the electrically conductive layer containing; a first redox mediator; and a reference electrode comprising; a non-conductive polymeric film, the polymeric film coated with an electrically conductive formulation comprising Ag/AgCl dispersed in a resin formulation, and with the reference electrode having an opening; and a reagent strip, the reagent strip comprising; a carrier strip that is a porous or fibrous water absorbent matrix, the carrier strip impregnated with a mixture comprising; glucose oxidase, horseradish peroxidase, a second redox mediator that can be oxidized by hydrogen peroxide under catalysis by horseradish peroxidase, at least one surfactant, at least one stabilizer, a buffering agent to maintain a pH from about 4 to about 8; and a whole blood separation membrane, the whole blood separation membrane comprising a porous matrix; with the porous matrix having an effective pore size of about 1 micron to about 10 microns; with the electrically conducting layer of the sensing electrode and the electrically conductive formulation of the reference electrode facing each other; and
with the reagent strip superimposed on and in physical contact with the electrically conducting layer of the sensing electrode, and with the whole blood separation membrane superimposed on and in physical contact with the reagent strip; and
with the reference electrode superimposed on the whole blood separation membrane;
so that the electrically conductive formulation coating the reference electrode is in physical contact with the whole blood separation membrane, and with the whole blood separation membrane completely covering the opening in the reference electrode;
so that the sensing electrode and the reagent strip and the whole blood separation membrane and the reference electrode form a sandwich;b) introducing a whole blood sample into the opening of the reference electrode; c) maintaining a potential of about -80 mV to about -125 mV across the sensing electrode and the reference electrode; d) measuring the current passing between the sensing electrode and the reference electrode; e) comparing the current measured to a calibration curve of the concentration of glucose versus current at the potential used in step c) to obtain the concentration of glucose in the whole blood sample. - View Dependent Claims (14, 15, 16, 17, 18, 19)
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20. A method of enhancing the sensitivity of an amperometric strip sensor for the assay of an analyte comprising the steps of:
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a) forming a sensing electrode, the sensing electrode comprising; a non-conductive support member, with the non-conductive support member comprising; a non-conductive polymeric film; the non-conductive polymeric film coated with an electrically conductive layer, with the electrically conductive layer containing; a first redox mediator; and b) forming a reference electrode comprising; a non-conductive polymeric film, the polymeric film coated with an electrically conductive formulation comprising Ag/AgCl dispersed in a resin formulation, c) providing an opening in either electrode for introduction of a sample; and d) forming a reagent strip, the reagent strip comprising; a carrier strip that is a porous or fibrous water absorbent matrix, the carrier strip impregnated with a mixture comprising; reagents that produce a flow of electrons when contacted by the analyte, at least one surfactant, at least one stabilizer, e) arranging the sensing electrode and the reference electrode and the reagent strip so that the electrically conductive layer of the sensing electrode and the electrically conductive formulation of the reference electrode face each other with the reagent strip superimposed on and in physical contact with the electrically conducting layer of the sensing electrode, and with the reference electrode superimposed on the reagent strip so that the electrically conductive formulation coating the reference electrode is in physical contact with the reagent strip;
so that the sensing electrode and the reagent strip and the reference electrode form a sandwich. - View Dependent Claims (21, 22)
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Specification