title: High-content analysis of kinase activity in cells.
abstract: High-content analysis (HCA) is a term used to describe techniques involving multiplexed analysis of fluorescent markers to measure multiple cellular responses to biological stimuli or drug treatment. HCA is usually based on automated microscopy or related technologies, and its value lies in providing multiparametric information on single cells within a population. During the last decade, several HCA approaches have been developed and applied to assess cellular mechanism of action of pharmacologically relevant compounds identified through biochemical screening or similar in vitro methods. With automation and instrument development, these approaches have evolved to the extent that the technique is now routinely used in screening applications, including primary HTS on compound collections. Here, we review the field and discuss in particular the application of HCA to the discovery of small molecule inhibitors targeting kinases which are implicated in Oncology.
authors.affiliation: Department of Cell Biology, Oncology Business Unit, Nerviano Medical Sciences Srl, Nerviano, Milano, Italy. fabio.gasparri@nervianoms.com
authors.forename: Fabio
authors.initials: F
authors.lastname: Gasparri
authors.forename: Francesco
authors.initials: F
authors.lastname: Sola
authors.forename: Tiziano
authors.initials: T
authors.lastname: Bandiera
authors.forename: Jürgen
authors.initials: J
authors.lastname: Moll
authors.forename: Arturo
authors.initials: A
authors.lastname: Galvani
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002477:Cells / Q000201:enzymology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005456:Fluorescent Dyes; D010770:Phosphotransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: D005456:Fluorescent Dyes; D054852:Small Molecule Libraries; D010770:Phosphotransferases