Patent ID: 7166475
Filing Date: 2007-01-23
Classification: C12Q,G01N

Abstract:
1. A method for detecting, in a sample, the presence of a modifying enzyme which covalently modifies a polypeptide, the method comprising the steps of: a) providing a polypeptide pair comprising a first polypeptide and a second, binding partner polypeptide, capable of associating with said first polypeptide, wherein said second, binding partner polypeptide is not a phospho-specific antibody, and wherein the association of the polypeptides is detectable, and covalent modification of at least one of the polypeptides results in modulation of the association and wherein said covalent modification comprises one of the group consisting of phosphorylation, acylation, glycosylation, ubiquitination, prenylation, sentrinization, and ADP-ribosylation; b) providing a modifying group substrate, wherein said substrate, in the presence of a modifying enzyme, results in the covalent modification of said first polypeptide or said second binding partner polypeptide; c) immobilizing the first polypeptide to a physical plasmon resonance support; d) contacting said immobilized polypeptide and said second binding partner polypeptide in the presence of said sample and said modifying group substrate; and e) measuring the association of the second, binding partner polypeptide to the first polypeptide, thereby determining the covalent modification of at least one of said polypeptides, whereby the presence of said modifying enzyme is detected; wherein said association is measured by monitoring the molecular mass of the binding partner by surface plasmon resonance.