High throughput screening technologies for direct cyclic AMP measurement.
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.
Gabriel
Daniela
D
Novartis Pharma AG, Lead Discovery Center, Basel, Switzerland. daniela.gabriel@pharma.novartis.com
Vernier
Magali
M
Pfeifer
Martin J
MJ
Dasen
Boris
B
Tenaillon
Laurent
L
Bouhelal
Rochdi
R
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
D011933:Reagent Kits, Diagnostic; D005576:Colforsin; D000242:Cyclic AMP; D019204:GTP-Binding Proteins