Method for continuous Immunoassay monitoring
First Claim
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1. A method for continuous determination of a target analyte in a sample comprising the steps of:
- (a) contacting an analyte with an immunosensory apparatus comprising a working electrode having a binder on its surface and a reference electrode and wherein said working electrode requires no regeneration between consecutive measurements of the analyte;
(b) determining a change in the working electrode potential, and (c) comparing the potentiometric response of the working electrode with the reference electrode wherein a difference indicates the presence of analyte in said sample and said difference is proportional to the concentration of analyte, and wherein said measurement can be performed with a single working electrode, thereby determining the concentration of said analyte in the sample.
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Abstract
The present invention relates to assays that employ an enzyme label or tag that acts on a substrate by obtaining electrons from an electrode (electrocatalysis) and an apparatus for use in such assays. In particular the present invention provides an assay and apparatus for multiple or continuous monitoring of the amount of analyte in a sample or sample source without requiring regeneration of the measuring electrode or its associated reagents.
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Citations
17 Claims
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1. A method for continuous determination of a target analyte in a sample comprising the steps of:
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(a) contacting an analyte with an immunosensory apparatus comprising a working electrode having a binder on its surface and a reference electrode and wherein said working electrode requires no regeneration between consecutive measurements of the analyte;
(b) determining a change in the working electrode potential, and (c) comparing the potentiometric response of the working electrode with the reference electrode wherein a difference indicates the presence of analyte in said sample and said difference is proportional to the concentration of analyte, and wherein said measurement can be performed with a single working electrode, thereby determining the concentration of said analyte in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, 17)
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14. A method for determining a target analyte in a sample comprising the steps of:
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(a) contacting an analyte with an immunosensory apparatus, comprising a diffusion membrane, a reference electrode and a working electrode, the latter having on its surface a binder with a labeled detection compound attached thereto, wherein said electrodes and said membrane are separated by a chamber containing a diffusion medium that contains a substrate of an electrocatalytic enzyme and wherein said electrodes are separated from the source of said analyte by said membrane, under conditions promoting diffusion of said analyte through said membrane to contact said electrodes;
(b) allowing the analyte to displace the labeled detection compound from said binder whereupon said analyte becomes bound to the binder on the surface of the working electrode in the case where the detection compound is the same as the analyte or the analyte becomes bound to the detection compound in the case where the labeled detection compound is a binder for the analyte, and wherein said detection compound is bound to an electrocatalytic enzyme such that in the presence of the target analyte the working electrode provides an electron to said electrocatalytic enzyme which provides an electron to a substrate of said enzyme (c) determining a change in the working electrode potential, and (d) comparing the potentiometric response of the working electrode with the reference electrode wherein a difference indicates the presence of analyte in said sample and is proportional to the concentration of analyte, thereby determining the concentration of analyte in the sample.
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Specification