title: Sensitivity improvement in 19F NMR-based screening experiments: theoretical considerations and experimental applications.
abstract: NMR-based binding and functional screening performed with FAXS (fluorine chemical shift anisotropy and exchange for screening) and 3-FABS (three fluorine atoms for biochemical screening) represents a potential alternative approach to high-throughput screening for the identification of novel potential drug candidates. The major limitation of this method in its current status is its intrinsic low sensitivity that limits the number of tested compounds. One approach for overcoming this problem is the use of a cryogenically cooled (19)F probe that reduces the thermal noise in the receiver circuitry. Sensitivity improvement in the two screening techniques achieved with the novel cryogenic (19)F probe technology permits an increased throughput, detection of weaker binders and inhibitors (relevant in a fragment-based lead discovery program), detection of slow binders, and reduction in protein and substrate consumption. These aspects are analyzed with theoretical simulations and experimental quantitative performance evaluation. Application of 3-FABS combined with the cryogenic (19)F probe technology to rapid screening at very low enzyme concentrations and the current detection limits reached with this approach are also presented.
authors.affiliation: Chemistry Department, Nerviano Medical Sciences, 20014 Nerviano, Milano, Italy. claudio.dalvit@nervianoms.com
authors.forename: Claudio
authors.initials: C
authors.lastname: Dalvit
authors.forename: Nicola
authors.initials: N
authors.lastname: Mongelli
authors.forename: Gianluca
authors.initials: G
authors.lastname: Papeo
authors.forename: Patrizia
authors.initials: P
authors.lastname: Giordano
authors.forename: Marina
authors.initials: M
authors.lastname: Veronesi
authors.forename: Detlef
authors.initials: D
authors.lastname: Moskau
authors.forename: Rainer
authors.initials: R
authors.lastname: Kümmerle
mesh_terms: D005461:Fluorine; D009682:Magnetic Resonance Spectroscopy / Q000379:methods* / Q000592:standards; D008962:Models, Theoretical; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D005461:Fluorine