text: In fragment-based drug discovery, the low binding affinities of the fragments pose significant challenges for screening. Many biophysical techniques have been applied to address this issue. In particular, ligand-observe nuclear magnetic resonance (NMR) methods such as water-ligand observed via gradient spectroscopy (waterLOGSY), saturation transfer difference spectroscopy (STD-NMR), F NMR spectroscopy and inter-ligand Overhauser effect (ILOE) spectroscopy, protein-observe NMR methods such as H-N heteronuclear single quantum coherence (HSQC) that utilises isotopically-labelled proteins, surface plasmon resonance (SPR), isothermal titration calorimetry (ITC) and Microscale Thermophoresis (MST) are routinely-used for ligand screening and for the quantification of fragment binding affinity to the target protein. At modern X-ray crystallography synchrotron beamlines, several hundred data sets of protein-ligand complex crystal structures can be obtained within 24 hours. This technology makes crystallographic fragment screening possible, i.e. the use of X-ray crystallography directly for the fragment screening step.
  Once a fragment (or a combination of fragments) have been identified, protein X-ray crystallography is used to obtain structural models of the protein- complexes. Such information can then be used to guide organic synthesis for high-affinity protein ligands and enzyme inhibitors.