Abstract: The invention relates to methods and reagents for identifying/isolating protein targets of chemical compounds (for example, drug candidates) using mass spectrometry. The invention provides a method for capturing and identifying proteins using tethered small-molecule probes. This technology also allows the market expansion of known drugs by finding new therapeutic targets; identification of the mechanism of toxicity of drug candidates or drugs which failed in the clinic; identification of new chemical tools for chemically-driven target validation; identification of new drug leads; and identification of the mechanism of action of drugs and drug candidates. A key advantage of the technology is that a single experiment can identify the numerous proteins which interact with a probe (or “bait”).

Abstract: A method and tools for the biochemical processing of protein samples towards their analysis and identification by, for example, mass spectrometry (MS). The method involves the reversible (non-covalent) immobilization, purification and concentration of proteins onto a solid surface and subsequent solid-phase enzyme-catalyzed proteolysis of the proteins. The constituent peptides are recovered in near-quantitative yields in a format ideally suited for identification by routine MS technologies. The process is easily adapted such that additional chemical and/or enzymatic transformation(s) of the immobilized proteins or peptides can be performed to facilitate the acquisition of and increase the information content obtained from the MS analysis.