Patent ID: 7632651
Filing Date: 2009-12-15
Classification: B01L,C07K,C12Q,G01N

Abstract:
1. A method of detecting the activity of an enzyme that catalyzes formation of a product from a substrate, the method comprising: contacting the substrate with the enzyme and with a binding partner to form a sample mixture, the binding partner binding selectively either to the substrate relative to the product or to the product relative to the substrate, the binding partner including at least one metal that is required for selective binding of the binding partner to the substrate or to the product via at least one phosphate moiety of the substrate or product, where the substrate and/or product includes an energy transfer donor and the binding partner includes an energy transfer acceptor, or the substrate and/or product includes an energy transfer acceptor and the binding partner includes an energy transfer donor, such that luminescence energy transfer can occur between the donor and the acceptor when the binding partner binds to the substrate or the product, where the enzyme is selected from the group consisting of kinases, phosphatases, nucleotide cyclases, and nucleotide phosphodiesterases, where the binding partner includes a lanthanide chelate and the metal, where the lanthanide chelate includes an organic chelator and a lanthanide, and where the organic chelator includes at least one metal-binding functional group that associates the lanthanide chelate with the metal; exposing the sample mixture to light capable of inducing luminescence from the energy transfer donor; measuring a detectable luminescence energy transfer response from the sample mixture, without separating the bound substrate or product from the unbound substrate or product, where the detectable luminescence energy transfer response is indicative of the extent of binding between the substrate or product and the binding partner; and correlating the response with the activity of the enzyme.