Fluorescent molecular probes for use in assays that measure test compound competitive binding with SAM-utilizing proteins

US9120820B2 · kind B2 · utility

Assignees

• CAYMAN CHEMICAL COMPANY, INC. · US
• THE REGENTS OF THE UNIVERSITY OF MICHIGAN · US

Inventors

• Stephen Douglas Barrett · Hartland, US
• Daniel Austin Bochar · Ann Arbor, US
• Levi BLAZER · Ann Arbor, US
• Fred Lawrence Ciske · Dexter, US
• Gregory William Endres · Saline, US
• Jeffrey Keith Johnson · Ann Arbor, US
• Gregory S. Keyes · Dexter, US
• Ranjinder Singh Sidhu · Ann Arbor, US
• Raymond Trievel · Ypsilanti, US
• Margaret Lynn Collins · Canton, US

Key dates

• Filing date: May 18, 2012
• Grant date: Sep 1, 2015
• Expiry date: Mar 5, 2033

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N2333/91011
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Assay methods may generally comprise forming homogeneous assay mixtures comprising target SAM-utilizing protein, fluorescent detection analyte, and test compound, incubating, and measuring FP or TR-FRET signal emitted in order to determine a measure of test compound-SAM -utilizing protein binding. Assay mixtures comprise a SAM-utilizing protein, and a fluorescent detection analyte that binds with the SAM-utilizing protein in the absence of test compound. Assay mixtures may further comprise a test compound. Assay mixture embodiments may generate FP or TR-FRET signal properties that are a function of the inherent binding interactions of both the test compound and the detection analyte with the SAM-utilizing protein. Fluorescent detection analytes comprise a fluorophore moiety, a covalent linker moiety, and a SAM-utilizing protein ligand moiety and could be utilized in FP or TR-FRET assays to measure test compound binding.