Fragment Hit Validation

ZoBio offers different approaches for biophysical fragment hit validation integrated with its core TINS fragment screening technology next to biochemical hit validation/characterization. We use orthogonal validation technologies which include SPR and protein observed NMR.

Surface Plasmon Resonance (SPR)
SPR as an orthogonal method is a powerful technique for hit validation. SPR is a highly complementary, sensitive, label free detection technology and together with TINS provide the perfect balance. ZoBio has created a unique fragment based program which utilizes TINS screening followed by SPR characterization. Using our exclusive TINS and Biacore T200 integrated platform allows us to further develop in the field of protein immobilization which in turn allows us to provide our customers with our very best expertise and knowledge.

Protein Observed NMR (HSQC)
Another approach to validating hits is using protein observed NMR to achieve efficient NMR data from the NMR spectroscopy of the protein that are extremely sensitive to ligand binding. This can be done when the target can be expressed in bacteria to incorporate stable NMR visible isotopes. This orthogonal technique can be used to validate and quantify binding as well as to define a low resolution binding site on the target.

Competition Binding
An alternative approach to fragment hit validation is competition binding studies. Competition binding studies can be carried out when known ligands (inhibitors) are available. Conducting a competition binding assay using TINS is a superior technique to validate hits. Using this technique means each hit is re-assayed as a singleton which in turn confirms binding and provides binding site information.

For more information or to discuss fragment hit validation please contact us directly.