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Comparison of the some of the most common methods used for hit confirmation.
| Technique | Specific requirements | Protein consumption | Throughput | Dynamic range | Information content |
|---|---|---|---|---|---|
| NMR | 15N labelling for 2D protein observed NMR | High | Medium | mM - μM | High (binding site) |
| SPR | Suitable immobilisation | Low | High | mM - pM | High (kinetics) |
| ITC | Protein and ligand in identical buffer | High | Low | mM - nM | High (thermodynamics) |
| MST | Fluorescent labelling of one partner | Low | Medium | mM - pM | Medium (affinity) |
