SpeedScreen: label-free liquid chromatography–mass spectrometry-based high-throughput screening for the discovery of orphan protein ligands

Chemogenomics: an emerging strategy for rapid target and drug discovery

“…Target-ligand interactions (so-called 'hits') are unambiguously identified in the absence of confounding variables. For example, fluorescent-based methods for detecting the LIGAND-INDUCED CONFORMATIONAL STABILIZATION of proteins 14 or MASS-SPECTROMETRY-based detection systems 15,16 have been described as a means to examine the effect of bound ligands. By contrast, cell-based and organismal assays -in which selected compounds are delivered directly to cells or organisms in vitro -identify hits within a relevant cellular context, but, because of the interaction with multiple targets, hits require additional mechanistic characterization.…”

Discovery of a Novel Mode of Protein Kinase Inhibition Characterized by the Mechanism of Inhibition of Human Mesenchymal-epithelial Transition Factor (c-Met) Protein Autophosphorylation by ARQ 197

“…The relative affinity of ARQ 197, and its c-Met inactive enantiomer ARQ 198, for the unphosphorylated c-Met protein and for a control type III RTK kinase domain (FGFR2) was measured by indirect affinity mass spectrometry (13). Briefly, binding mixtures were 25 l in volume and contained 14 M protein, 20 M inhibitor in 25 mM Tris-HCl, pH 7.5, 100 mM NaCl, 0.1% 2-mercaptoethanol, and a 2% final DMSO concentration.…”

A Chemoinformatics Analysis of Hit Lists Obtained from High-Throughput Affinity-Selection Screening

“…1,2 SpeedScreen has become one of the most frequently used HTS platforms at Novartis, especially when genomic targets with an unknown function (orphan targets) or targets with a known function but nontractable to a functional HTS have to be addressed. To achieve HTS performance despite the sequential MS readout, a primary SpeedScreen campaign is performed with pools of 400 compounds per well of the 96-well microplates.…”