Patent ID: 6872574

Claim:
A method for screening for the bloactivity of a candidate compound toward a group of related target proteins in a proteomic mixture of proteins from a cell, employing at least one probe, each probe comprising a reactive functionality group specific for said group of target proteins and a ligand, each probe of the formula: R(F-L)—X wherein: X is a ligand for binding to a reciprocal receptor and/or providing a detectable signal; L is an alkylene, oxyalkylene or polyoxyalkylene linking group, wherein said oxyalkylenes are of from 2 to 3 carbon atoms; F is a phosphonate or sulfonyl functional group reactive at an active site of a target enzyme; and R is bonded to F and a moiety of less than 1 kDal providing specific affinity for said enzymes, and when F is phosponate, R is fluorine and when F is sulfonyl, R is an aryl or heteroaryl group; said method comprising: separating the proteomic mixture into two portions, treating one of the two portions with the candidate compound to obtain an inactivated portion, wherein the candidate compound is a non-covalent agent, combining at least one probe with the untreated portion of said mixture under conditions for reaction with said target proteins to obtain a first specimen, combining at least one probe with the inactivated portion under conditions for reaction with said target proteins to obtain a second specimen; sequestering proteins conjugated with said at least one probe from each of said specimens; determining the proteins that are sequestered; and comparing the amount of each of the proteins sequestered from the first specimen and the second specimen as indicative of the bioactivity of said candidate compound with said target proteins.