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SBDD/FBDD (FBDD) – cont’d

X-ray crystallography-based FBDD
Mass spectrometry based FBDD

Protein NMR Spectroscopy
    Features 

      Solution binding
      Structure information and binding mode
      Measurement of weak binding   

      Relatively low throughput

      Requires high purity isotope labeled protein

       Requires high compound concentration
 

SPR based technologies
    Features 
      Well established methods
      Can measure kinetics of binding event (on and off rate)
      Throughput can be high (with more expensive multichannel instruments, 96-well based)
      Can detect low affinity binding
      Requires protein immobilization (method development and validation)
      Throughput can be low (with less expensive instruments)
      Problem with low MW compounds (especially with older instruments)
 

Thermo-shift binding assay

Measurement of changes of protein thermo melting curve upon ligand binding
    Features 
      Solution binding, no protein labeling required
      Reasonably high throughput (96- and 384-well based)
      Applicable to many proteins
      Requires well-behaved protein thermo-denaturation curve
      Requires method development and validation for each protein
  

Other Methods (calorimetry, activity based methods, etc.)