One-step enzymatic and amine detection technique
First Claim
1. A method for detecting an amine, enzyme, or enzyme inhibitor within a test sample, the method comprising:
- i) contacting an incubation mixture that comprises the test sample that may contain an amine, enzyme, or enzyme inhibitor and further comprises a plurality of reactive complexes that each comprises a substrate joined to a reporter and a specific binding member, said substrate being cleavable by a hydrolytic enzyme to release said reporter with a chromatographic medium, said chromatographic medium defining an enzyme detection zone within which is immobilized a receptive material that has an affinity for said specific binding member for generating an enzyme detection signal wherein the presence or quantity of a hydrolytic enzyme is determinable from said enzyme detection signal, and an amine detection zone positioned downstream from said enzyme detection zone, wherein a chemichromic dye is contained within said amine detection zone, said chemichromic dye being capable of undergoing a color change in the presence of an amine, wherein the presence or quantity of an amine is determinable from said color change;
ii) determining the presence or quantity of an enzyme or enzyme inhibitor from said enzyme detection signal; and
iii) determining the presence or quantity of an amine from said amine detection signal.
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Abstract
A technique for detecting the presence or quantity of an enzyme (or enzyme inhibitor) and/or an amine within a test sample is provided. For example, in one embodiment, a diagnostic test kit is employed that utilizes reactive complexes that each includes a substrate joined (e.g., covalently bonded, physically adsorbed, etc.) to a reporter and a separation species. Upon contacting the reactive complexes, enzymes may cleave the substrate and release the reporter. Moreover, the test kit may also employ a chemichromic dye, i.e., a dye that exhibits a detectable color change upon chemical reaction with one or more functional groups, such as amino groups. The signal generated (directly or indirectly) by the reporter and chemichromic dye may then be used to indicate the presence or quantity of an enzyme (or enzyme inhibitor) and amine, respectively, within the test sample.
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Citations
7 Claims
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1. A method for detecting an amine, enzyme, or enzyme inhibitor within a test sample, the method comprising:
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i) contacting an incubation mixture that comprises the test sample that may contain an amine, enzyme, or enzyme inhibitor and further comprises a plurality of reactive complexes that each comprises a substrate joined to a reporter and a specific binding member, said substrate being cleavable by a hydrolytic enzyme to release said reporter with a chromatographic medium, said chromatographic medium defining an enzyme detection zone within which is immobilized a receptive material that has an affinity for said specific binding member for generating an enzyme detection signal wherein the presence or quantity of a hydrolytic enzyme is determinable from said enzyme detection signal, and an amine detection zone positioned downstream from said enzyme detection zone, wherein a chemichromic dye is contained within said amine detection zone, said chemichromic dye being capable of undergoing a color change in the presence of an amine, wherein the presence or quantity of an amine is determinable from said color change; ii) determining the presence or quantity of an enzyme or enzyme inhibitor from said enzyme detection signal; and iii) determining the presence or quantity of an amine from said amine detection signal. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification