title: Target immobilization as a strategy for NMR-based fragment screening: comparison of TINS, STD, and SPR for fragment hit identification.
abstract: Fragment-based drug discovery (FBDD) has become a widely accepted tool that is complementary to high-throughput screening (HTS) in developing small-molecule inhibitors of pharmaceutical targets. Because a fragment campaign can only be as successful as the hit matter found, it is critical that the first stage of the process be optimized. Here the authors compare the 3 most commonly used methods for hit discovery in FBDD: high concentration screening (HCS), solution ligand-observed nuclear magnetic resonance (NMR), and surface plasmon resonance (SPR). They selected the commonly used saturation transfer difference (STD) NMR spectroscopy and the proprietary target immobilized NMR screening (TINS) as representative of the array of possible NMR methods. Using a target typical of FBDD campaigns, the authors find that HCS and TINS are the most sensitive to weak interactions. They also find a good correlation between TINS and STD for tighter binding ligands, but the ability of STD to detect ligands with affinity weaker than 1 mM K(D) is limited. Similarly, they find that SPR detection is most suited to ligands that bind with K(D) better than 1 mM. However, the good correlation between SPR and potency in a bioassay makes this a good method for hit validation and characterization studies.
authors.affiliation: ZoBio B.V, Einsteinweg 55, 2333CC, Leiden, The Netherlands.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Kim
authors.initials: K
authors.lastname: Retra
authors.forename: Francis
authors.initials: F
authors.lastname: Figaroa
authors.forename: Johan G
authors.initials: JG
authors.lastname: Hollander
authors.forename: Eiso
authors.initials: E
authors.lastname: Ab
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Heetebrij
authors.forename: Hubertus
authors.initials: H
authors.lastname: Irth
authors.forename: Gregg
authors.initials: G
authors.lastname: Siegal
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D008024:Ligands; D008954:Models, Biological; D058990:Molecular Targeted Therapy; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000032:analysis; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D055767:Immobilized Proteins; D008024:Ligands; D010446:Peptide Fragments; D019151:Peptide Library; D054852:Small Molecule Libraries