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X-ray crystallography is an established technique for ligand screening in fragment-based drug-design projects, but the required manual handling steps - soaking crystals with ligand and the subsequent harvesting - are tedious and limit the throughput of the process. Here, an alternative approach is reported: crystallization plates are pre-coated with potential binders prior to protein crystallization and X-ray diffraction is performed directly `in situ' (or in-plate). Its performance is demonstrated on distinct and relevant therapeutic targets currently being studied for ligand screening by X-ray crystallography using either a bending-magnet beamline or a rotating-anode generator. The possibility of using DMSO stock solutions of the ligands to be coated opens up a route to screening most chemical libraries.
Supporting information
PDB references: human nuclear receptor PPARγ, complex with rosiglitazone, 4xld; hen egg-white lysozyme, complex with benzamidine, 4xn6; human cyclophilin D, complex with ethyl 2-({\[(4-aminophenyl)methyl\]carbamoyl}amino)acetate, room temperature, 4xnc; 4zsc; 100 K, 3rdc; complex with 1-(4-aminobenzyl)-3-\[4-(methylthio)-1-{2-\[2-(methylthio)\]phenyl}pyrrolidin-1-yl)-1-oxobutan-2-yl\]urea, 100 K, 4j5c; room temperature, 4zsd; Erk-2, complex with 1-phenyl-1H-1,2,4-triazole-3,5-diamine, room temperature, 4xne; 100 K, 4xp2; complex with 2-amino-6-thiopurine, room temperature, 4xoy; 100 K, 4xp3; complex with 1-N-{\[3-(benzyloxy)phenyl\]methyl}-2H-1,2,3,4-tetrazole-1,5-diamine, room temperature, 4xrj; 100 K, 4xoz; complex with 3-cyano-7-azaindole, room temperature, 4xrl; 100 K, 4xp0
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