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), wherein said EAM comprises a transition metal complex and an enzyme reactive moiety (ERM) and has a first E°
, and wherein said ERM is removed from at least a portion of said EAM in the presence of at least one target analyte in the test sample and results in said EAM having a second E0;
(b) 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 an assay mixture in a solution phase is formed in step (a) and prior to step (b), and further comprising contacting said assay mixture with a solid support comprising an electrode, under conditions such that a self assembled monolayer (SAM) forms on said electrode.
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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, E01 and E02 that is detected through a self-assembled monolayer (SAM) on an electrode.
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Citations
19 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), wherein said EAM comprises a transition metal complex and an enzyme reactive moiety (ERM) and has a first E°
, and wherein said ERM is removed from at least a portion of said EAM in the presence of at least one target analyte in the test sample and results in said EAM having a second E0;(b) 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 an assay mixture in a solution phase is formed in step (a) and prior to step (b), and further comprising contacting said assay mixture with a solid support comprising an electrode, under conditions such that a self assembled monolayer (SAM) forms on said electrode. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. 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 E0 and comprising a transition metal complex and an enzyme reactive moiety (ERM), wherein said ERM is removed from at least a portion of said EAM in the presence of at least one target analyte in the test sample and results in said EAM having a second E0; (b) contacting said assay mixture with a solid support comprising an electrode under conditions such that a self-assembled monolayer (SAM) forms on said electrode, said SAM comprising said EAM having said first E0 and said EAM having said second E0; and (c) 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.
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Specification