title: Miniaturization and automation of an ubiquitin ligase cascade enzyme-linked immunosorbent assay in 1,536-well format.
abstract: Enzyme-linked immunosorbent assays (ELISAs) are a long established and widely used assay format for drug discovery and diagnostics. They offer many advantages over homogeneous assay formats, including high sensitivity and separation (wash) steps that remove detection-interfering compounds. Many high-throughput screening assays are now performed in miniaturized formats (1,536- and 3,456-well plates) for higher throughput and lower reagent consumption. With miniaturization, separation steps in assays such as ELISA can become difficult to implement. Here we report on the implementation of the Kalypsys, Inc. (San Diego, CA) 1,536-well plate washer to enable the successful miniaturization and full automation of an ELISA that monitors ubiquitin ligase activity. The 1,536-well plate ELISA was robust and used for the high-throughput screening of a large screening collection (>1 million compounds).
authors.affiliation: Department of Automated Biotechnology, Merck Research Laboratories, North Wales, PA 19454, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Cassaday
authors.forename: Tarak
authors.initials: T
authors.lastname: Shah
authors.forename: Justin
authors.initials: J
authors.lastname: Murray
authors.forename: Gregory T
authors.initials: GT
authors.lastname: O'Donnell
authors.forename: Oleg
authors.initials: O
authors.lastname: Kornienko
authors.forename: Berta
authors.initials: B
authors.lastname: Strulovici
authors.forename: Marc
authors.initials: M
authors.lastname: Ferrer
authors.forename: Paul
authors.initials: P
authors.lastname: Zuck
mesh_terms: D001331:Automation; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D008904:Miniaturization; D012371:Robotics; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry*
chemical_list: D044767:Ubiquitin-Protein Ligases