title: Fluorescent biosensors for high throughput screening of protein kinase inhibitors.
abstract: High throughput screening assays aim to identify small molecules that interfere with protein function, activity, or conformation, which can serve as effective tools for chemical biology studies of targets involved in physiological processes or pathways of interest or disease models, as well as templates for development of therapeutics in medicinal chemistry. Fluorescent biosensors constitute attractive and powerful tools for drug discovery programs, from high throughput screening assays, to postscreen characterization of hits, optimization of lead compounds, and preclinical evaluation of candidate drugs. They provide a means of screening for inhibitors that selectively target enzymatic activity, conformation, and/or function in vitro. Moreover, fluorescent biosensors constitute useful tools for cell- and image-based, multiplex and multiparametric, high-content screening. Application of fluorescence-based sensors to screen large and complex libraries of compounds in vitro, in cell-based formats or whole organisms requires several levels of optimization to establish robust and reproducible assays. In this review, we describe the different fluorescent biosensor technologies which have been applied to high throughput screens, and discuss the prerequisite criteria underlying their successful application. Special emphasis is placed on protein kinase biosensors, since these enzymes constitute one of the most important classes of therapeutic targets in drug discovery.
authors.affiliation: CRBM-CNRS-UMR 5237, Chemical Biology and Nanotechnology for Therapeutics, 1919 Route de Mende, 34293 Montpellier, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Prével
authors.forename: Morgan
authors.initials: M
authors.lastname: Pellerano
authors.forename: Thi Nhu Ngoc
authors.initials: TN
authors.lastname: Van
authors.forename: May C
authors.initials: MC
authors.lastname: Morris
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D005456:Fluorescent Dyes; D057166:High-Throughput Screening Assays; D006801:Humans; D051379:Mice; D047428:Protein Kinase Inhibitors
chemical_list: D005456:Fluorescent Dyes; D047428:Protein Kinase Inhibitors
keywords: Drug Discovery; Fluorescent Biosensor; High Throughput Screening; Inhibitor; Protein Kinase