title: Fragment-Based Screening for Enzyme Inhibitors Using Calorimetry.
abstract: Isothermal titration calorimetry (ITC) provides a sensitive and accurate means by which to study the thermodynamics of binding reactions. In addition, it enables label-free measurement of enzymatic reactions. The advent of extremely sensitive microcalorimeters have made it increasingly valuable as a tool for hit validation and characterization, but its use in primary screening is hampered by requiring large quantities of reagents and long measurement times. Nanocalorimeters can overcome these limitations of conventional ITC, particularly for screening libraries of 500-1000 compounds such as those encountered in fragment-based lead discovery. This chapter describes how nanocalorimetry and conventional microcalorimetry can be used to screen compound libraries for enzyme inhibitors.
authors.affiliation: Palo Alto Research Center, Palo Alto, California, USA. Electronic address: michael.recht@parc.com.
authors.forename: Michael I
authors.initials: MI
authors.lastname: Recht
authors.affiliation: Zenobia Therapeutics, San Diego, California, USA.
authors.forename: Vicki
authors.initials: V
authors.lastname: Nienaber
authors.affiliation: Palo Alto Research Center, Palo Alto, California, USA.
authors.forename: Francisco E
authors.initials: FE
authors.lastname: Torres
mesh_terms: D002151:Calorimetry / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors
chemical_list: D004791:Enzyme Inhibitors
keywords: Enzyme assay; Fragment library; Fragment-based lead discovery; Label-free assay; Microcalorimetry; Nanocalorimetry