title: High throughput screening technologies for direct cyclic AMP measurement.
abstract: A study comparing five different cAMP detection technologies in terms of sensitivity, robustness, and feasibility for HTS is presented. In this report, the following methods are described: a nonhomogeneous DELFIA, and the homogeneous methods based on time-resolved fluorescence (HTRF), luminescent singlet oxygen channeling or ALPHAScreen, FP, and high-affinity enzyme complementation. DELFIA had the highest sensitivity, whereas ALPHAScreen and HTRF shared several advantages, including high sensitivity, broad dynamic range, and minimal reagent addition steps. For G(s)-coupled antagonist screens, we found HTRF and ALPHAScreen the more sensitive and HTS-compatible techniques.
authors.affiliation: Novartis Pharma AG, Lead Discovery Center, Basel, Switzerland. daniela.gabriel@pharma.novartis.com
authors.forename: Daniela
authors.initials: D
authors.lastname: Gabriel
authors.forename: Magali
authors.initials: M
authors.lastname: Vernier
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Pfeifer
authors.forename: Boris
authors.initials: B
authors.lastname: Dasen
authors.forename: Laurent
authors.initials: L
authors.lastname: Tenaillon
authors.forename: Rochdi
authors.initials: R
authors.lastname: Bouhelal
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D016466:CHO Cells; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D000242:Cyclic AMP / Q000032:analysis* / Q000096:biosynthesis; D005453:Fluorescence; D015200:Fluoroimmunoassay / Q000379:methods; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008163:Luminescent Measurements; D011933:Reagent Kits, Diagnostic; D013678:Technology, Pharmaceutical
chemical_list: D011933:Reagent Kits, Diagnostic; D005576:Colforsin; D000242:Cyclic AMP; D019204:GTP-Binding Proteins