Combining 'dry' co-crystallization and in situ diffraction to facilitate ligand screening by X-ray crystallography

Acta Crystallogr D Biol Crystallogr. 2015 Aug;71(Pt 8):1777-87. doi: 10.1107/S1399004715010342. Epub 2015 Jul 31.


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.

Keywords: drug design; fragment library screening; in-situ X-ray diffraction; ligand screening; therapeutic targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chickens
  • Crystallization / methods*
  • Crystallography, X-Ray / methods*
  • Cyclophilin D
  • Cyclophilins / chemistry
  • Cyclophilins / metabolism
  • Drug Discovery / methods*
  • Humans
  • Ligands
  • Mitogen-Activated Protein Kinase 1 / chemistry
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Muramidase / chemistry
  • Muramidase / metabolism
  • PPAR gamma / chemistry
  • PPAR gamma / metabolism
  • Proteins / chemistry*
  • Proteins / metabolism
  • Rats


  • Cyclophilin D
  • Ligands
  • PPAR gamma
  • Proteins
  • Mitogen-Activated Protein Kinase 1
  • hen egg lysozyme
  • Muramidase
  • Cyclophilins