ELECTRIC MEASUREMENT OF MONOLAYERS FOLLOWING PRO-CLEAVE DETECTION OF PRESENCE AND ACTIVITY OF ENZYMES AND OTHER TARGET ANALYTES
First Claim
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1. A method for detecting at least one target analyte in a test sample, said method comprising:
- (a) contacting a test sample with an electroactive moiety (EAM), said EAM comprising a transition metal complex and an enzyme reactive moiety (ERM) and having a first Eo, under conditions wherein said ERM is removed from at least a portion of said EAM provided in the presence of at least one target analyte in the test sample and results in said EAM having a second Eo;
(b) detecting for a change between the first Eo and the second Eo of said EAM, wherein said change is an indication of the presence of said at least one target analyte.
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Abstract
This invention describes methods for detecting target analytes by utilizing an electroactive moiety (EAM) that functions as a substrate for a specific enzyme. If target analyte is present, a functional group is enzymatically removed from a transition metal complex resulting in quantifiable electrochemical signal at two unique potentials, Eo1 and Eo2 that is detected through a self-assembled monolayer (SAM) on an electrode.
21 Citations
30 Claims
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1. A method for detecting at least one target analyte in a test sample, said method comprising:
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(a) contacting a test sample with an electroactive moiety (EAM), said EAM comprising a transition metal complex and an enzyme reactive moiety (ERM) and having a first Eo, under conditions wherein said ERM is removed from at least a portion of said EAM provided in the presence of at least one target analyte in the test sample and results in said EAM having a second Eo; (b) detecting for a change between the first Eo and the second Eo of said EAM, wherein said change is an indication of the presence of said at least one target analyte. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. A composition comprising a solid support comprising:
an electrode comprising; (i) a self-assembled monolayer (SAM); and (ii) a covalently attached electroactive active moiety (EAM) comprising a transition metal complex and an enzyme reactive moiety (ERM), wherein said EAM has a first E0 when said ERM is present and a second E0 when said ERM is absent. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23)
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24. A method for detecting at least one target analyte in a test sample, said method comprising:
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(a) contacting a test sample with an electroactive moiety (EAM) to form an assay mixture in solution phase, said EAM having a first Eo and comprising a transition metal complex and an enzyme reactive moiety (ERM), under conditions wherein said ERM is removed from at least a portion of said EAM provided in the presence of at least one target analyte in the test sample and results in said EAM having a second Eo; (b) contacting said assay mixture with a solid support comprising an electrode or an array of electrodes under conditions such that a self-assembled monolayer (SAM) forms on said electrode, said SAM comprising said EAM having said first Eo and said EAM having said second Eo; and (c) detecting for a change between the first Eo and the second Eo of said EAM, wherein said change is an indication of the presence of said at least one target analyte.
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25. A method for detecting at least one target analyte in a test sample, said method comprising:
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(a) contacting a test sample with a solid support, said solid support comprising an electrode or array of electrodes, said electrode comprising; (i) a self-assembled monolayer; and (ii) a covalently attached electroactive moiety (EAM), said EAM having a first Eo and comprising a transition metal complex and an enzyme reactive moiety (ERM), under conditions wherein said ERM is removed from at least a portion of said EAM provided in the presence of at least one target analyte in the test sample and results in said EAM having a second Eo; (b) detecting for a change between the first Eo and the second Eo of said EAM, wherein said change is an indication of the presence of said at least one target analyte.
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26. A method for detecting at least one target analyte in a test sample, said method comprising:
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(a) contacting a test sample with a capture binding ligand bound to a first solid support under conditions so as to form a first complex in the presence of at least one target analyte; (b) contacting the first complex with a soluble binding ligand comprising a surrogate enzyme under conditions so as to form a second complex on said first solid support; (c) contacting the second complex with an electroactive moiety (EAM), said EAM comprising a transition metal complex and an enzyme reactive moiety (ERM) and having a first E0, under conditions wherein said ERM is removed from at least a portion of said EAM provided in the presence of at least one target analyte in the test sample and results in said EAM having a second E0; and (d) detecting for a change between the first E0 and the second E0 of said EAM, wherein said change is an indication of the presence of said at least one target analyte, wherein said surrogate enzyme is an enzyme that removes said ERM from said EAM. - View Dependent Claims (27, 28, 29, 30)
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Specification