Fragment library screening using TINS (Target Immobilized NMR Screening)
ZoBio offers its proprietary TINS technology for ligand screening studies on its own, highly validated fragment library or the customer’s. TINS uses a single sample of the target and a reference protein that have been immobilized on sepharose based resin. Fragments to be assayed for binding are injected as mixtures of between 3 and 9 compounds. Binding is detected by a reduction in the height of the NMR signals of a compound that specifically binds to the target. Comparison of the NMR spectra of the fragments in the presence of the target and reference proteins directly reveals the identity of a ligand. As shown below, the screen is carried out by repeated cycles of fragment application, assaying for binding and fragment removal. TINS is a powerful approach to hit discovery that has been validated on a wide range of targets including viral proteins, proteases, kinases, Protein-Protein Interaction targets, molecular chaperones and membrane proteins such as GPCRs. A typical TINS hit discovery and hit characterization/validation project is completed in 1-2 months.
Key TINS features
Applicable to a broad range of targets, including unstable proteins, proteases and membrane proteins
Requires only a single sample of the target, between 10-20 nmoles
All targets functionally immobilized using various conjugation techniques
Comparative technique, i.e. use of a reference protein: extremely high level of ligand specificity
Direct detection of binding, no deconvolution required
Allows for ‘blind’ screening, no known ligand a priori required
Good hit rates due to exceptional sensitivity, low false positive rate
Generates lots of unique starting compounds that show high validation rate
Compounds from screens are directly amenable to further chemistry
Fast: 1,500 compounds screened in 3-4 days, complete hit discovery project finished within 1-2 months
See the following publications for more details:
Vanwetswinkel, et al, "TINS: Target Immobilized NMR Screening – An efficient and sensitive new method for ligand discovery", Chem. Biol., 12, 2005, pp.207
Marquardsen et al, "Development of a dual cell, flow-injection sample holder, and NMR probe for comparative ligand-binding studies", J. Mag. Reson., 2006, 182, pp. 55
Siegal, G., AB, Eiso and Schultz, J., "Integration of Fragment Screening and Library Design", Drug Discovery Today, 2007, 12, p.1032-1039
Siegal,G. and Hollander,J.G., "Target Immobilization and NMR Screening of Fragments in Early Drug Discovery",Current Topics Med. Chem., 2009, 9, p.173
Kobayashi et al, "Target Immobilization as a Strategy for NMR-Based Fragment Screening: Comparison of TINS, STD, and SPR for Fragment Hit Identification", J. Biomol. Screen., 2010, 15, p.978
Früh, V., "Application of Fragment-Based Drug Discovery to Membrane Proteins: Identification of Ligands of the Integral Membrane Enzyme DsbB", Chem. & Biol., 2010, 17, p. 881
Congreve,M. et al, "Fragment Screening of Stabilized G-Protein Coupled Receptors using Biophysical Methods", Methods in Enzymology, 2011, 493, p.115
