Assay for proline iminopeptidase and other hydrolytic activities
First Claim
1. A method for assaying for the presence or absence of an analyte selected from the group consisting of a catalytically active hydrolase and an inhibitor of a catalytically active hydrolase in a sample, said method comprising:
- (a) contacting said sample with a solid-phase conjugate consisting of a reporter group coupled to a substrate residue yet cleavable therefrom by said catalytically active hydrolase, said reporter group when not so coupled being capable of causing a detectable change in an indicator, this step being performed in an environment in which the condition of said reporter group as either coupled or decoupled correlates with the presence or absence of said analyte;
(b) during or subsequent to step (a), contacting said sample with a solid-phase indicator which undergoes a detectable change upon action of said reporter group; and
(c) observing whether said indicator undergoes a detectable change, said detectable change being an indication of the presence or absence of said analyte in said sample.
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Accused Products
Abstract
The presence of an enzymatically active hydrolase in a fluid sample is detected by contacting the sample with a solid-phase conjugate which is susceptible to cleavage by the hydrolase, and simultaneously or shortly thereafter, contacting the sample with an indicator which undergoes a detectable change upon the action of a reporter group. The reporter group is part of the conjugate and is liberated from it either partly or entirely by the action of the hydrolase. The indicator is susceptible to action by the reporter group only upon decoupling of the reporter group from the remainder of the conjugate, the decoupling occurring either in part or entirely upon action of the hydrolase. Also provided by this invention are various forms of a dry, self-contained test device which contains the conjugate described above plus the indicator and all other reagents and components necessary to achieve a detectable indication of the presence or absence of a catalytically active hydrolase. Preferred embodiments of the device also contain positive and negative controls.
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Citations
48 Claims
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1. A method for assaying for the presence or absence of an analyte selected from the group consisting of a catalytically active hydrolase and an inhibitor of a catalytically active hydrolase in a sample, said method comprising:
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(a) contacting said sample with a solid-phase conjugate consisting of a reporter group coupled to a substrate residue yet cleavable therefrom by said catalytically active hydrolase, said reporter group when not so coupled being capable of causing a detectable change in an indicator, this step being performed in an environment in which the condition of said reporter group as either coupled or decoupled correlates with the presence or absence of said analyte; (b) during or subsequent to step (a), contacting said sample with a solid-phase indicator which undergoes a detectable change upon action of said reporter group; and (c) observing whether said indicator undergoes a detectable change, said detectable change being an indication of the presence or absence of said analyte in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A test device for assaying for the presence of a catalytically active hydrolase in a sample, said test device comprising:
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a receptacle defined at least in part by first and second opposing walls having interior-facing surfaces with a gap therebetween, said first wall, said second wall, or both being of light-transmitting material; a solid-phase conjugate deposited on said interior-facing surface of one of said first and second walls, said conjugate consisting of a reporter group coupled to a substrate residue yet cleavable therefrom upon contact with said catalytically active hydrolase, said reporter group when not so coupled being capable of causing a detectable change in an indicator; a solid-phase indicator deposited on said interior-facing surface of one of said first and second walls, said indicator being one which undergoes a detectable change upon action of said reporter group; and an opening in said receptacle for introduction of said sample. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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35. A test device for assaying for the presence of an analyte in a sample, said test device comprising:
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a penetrable solid support defined at least in part by upper and lower surfaces; a reagent contained in a solid layer on one of said upper and lower surfaces of said penetrable solid support, said reagent being one which induces a chemical reaction in an indicator when said analyte is present in said sample; and an indicator contained in a solid layer on the other of said upper and lower surfaces of said penetrable solid support in a manner such that said indicator is separated from said reagent until said sample is added, said indicator being one which undergoes a detectable change upon the occurrence of said chemical reaction. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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46. A test device for assaying for the presence of an analyte in a liquid sample, said test device comprising:
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a solid support having a surface; a solid-phase reagent deposited in a first region on said surface, said reagent being one which induces a chemical reaction in an indicator when said analyte is present in said sample; a solid-phase indicator deposited in a second region on said surface, said second region non-overlapping with said first region, said indicator being one which undergoes a detectable change upon the occurrence of said chemical reaction; and means for drawing said sample across said surface from said first region to said second region. - View Dependent Claims (47, 48)
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Specification