Methods of detecting target molecules and molecular interactions
First Claim
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1. A method of detecting a target molecule in a sample comprising the steps of;
- (a) contacting a sample with two or more binding entities specific for the target molecule, including a first binding entity labelled with a catalytic label and a second, separate binding entity labelled with a substrate label, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label;
(b) incubating under conditions which permit the binding entities to bind to the target molecule, thus bringing the catalytic and substrate labels into close proximity;
(c) allowing the catalytic label to act directly on the substrate label when bound in close proximity, thereby producing a detectable change in the substrate label;
(d) detecting the change in the substrate label.
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Abstract
Methods and kits for detecting target molecules or for monitoring interactions between target molecules. In particular, the invention relates to methods based on the use of “two-component conjugate” systems comprising a catalytic label and a substrate label.
25 Citations
34 Claims
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1. A method of detecting a target molecule in a sample comprising the steps of;
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(a) contacting a sample with two or more binding entities specific for the target molecule, including a first binding entity labelled with a catalytic label and a second, separate binding entity labelled with a substrate label, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label;
(b) incubating under conditions which permit the binding entities to bind to the target molecule, thus bringing the catalytic and substrate labels into close proximity;
(c) allowing the catalytic label to act directly on the substrate label when bound in close proximity, thereby producing a detectable change in the substrate label;
(d) detecting the change in the substrate label. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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2. A method of detecting interactions between two or more interacting molecules comprising the steps of:
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(a) incubating the interacting molecules such that they can interact, thereby bringing into close proximity a catalytic label attached to one of the interacting molecules and a substrate label attached to a separate interacting molecule, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label;
(b) allowing the catalytic label to act directly on the substrate label when bound in close proximity, thereby producing a detectable change in the substrate label;
(c) detecting the change in the substrate label. - View Dependent Claims (31)
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32. A reagent kit comprising a first binding entity labelled with a catalytic label and a second binding entity labelled with a substrate label, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label.
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33. A reagent kit comprising a first interacting molecule labelled with a catalytic label and a second interacting molecule labelled with a substrate label, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label.
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34. A reagent labelling kit comprising a catalytic label and a substrate label, characterised in that the catalytic label is capable of acting directly on the substrate label to generate a detectable change in the substrate label, and means for attaching the catalytic label and substrate label to interacting molecules or to binding entities.
Specification