title: Enabling 1536-Well High-Throughput Cell-Based Screening through the Application of Novel Centrifugal Plate Washing.
abstract: Cell-based assays have long been important within hit discovery paradigms; however, improving the disease relevance of the assay system can positively affect the translation of small-molecule drug discovery, especially if adopted in the initial hit identification assay. Consequently, there is an increasing need for disease-relevant assay systems capable of running at large scale, including the use of induced pluripotent stem cells and donor-derived primary cells. Major hurdles to adopting these assays for high-throughput screening are the cost, availability of cells, and complex protocols. Miniaturization of such assays to 1536-well format is an approach that can reduce costs and increase throughput. Adaptation of these complex cell assays to 1536-well format brings major challenges in liquid handling for high-content assays requiring washing steps and coating of plates. In addition, problematic edge effects and reduced assay quality are frequently encountered. In this study, we describe the novel application of a centrifugal plate washer to facilitate miniaturization of a range of 1536-well cell assays and techniques to reduce edge effects, all of which improved throughput and data quality. Cell assays currently limited in throughput because of cost and complex protocols may be enabled by the techniques presented in this study.
authors.affiliation: 1 Discovery Sciences, AstraZeneca, Cambridge, UK.
authors.forename: Sinead
authors.initials: S
authors.lastname: Knight
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Helen
authors.initials: H
authors.lastname: Plant
authors.affiliation: 1 Discovery Sciences, AstraZeneca, Cambridge, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: McWilliams
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: David
authors.initials: D
authors.lastname: Murray
authors.affiliation: 3 Department of Biochemistry, University of Bristol.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Dixon-Steele
authors.affiliation: 4 Quality Operations, Sanofi, Holmes Chapel, Cheshire, UK.
authors.forename: Anet
authors.initials: A
authors.lastname: Varghese
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Harper
authors.affiliation: 5 Discovery Sciences, AstraZeneca, Gothenburg, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Ramne
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Paula
authors.initials: P
authors.lastname: McArdle
authors.affiliation: 5 Discovery Sciences, AstraZeneca, Gothenburg, Sweden.
authors.forename: Susanna
authors.initials: S
authors.lastname: Engberg
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Bennett
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Blackett
authors.affiliation: 2 Discovery Sciences, AstraZeneca, Macclesfield, Cheshire, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Wigglesworth
mesh_terms: D000818:Animals; D015415:Biomarkers; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D055808:Drug Discovery / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods; D010641:Phenotype
chemical_list: D015415:Biomarkers
keywords: 1536-well; cell assays; high-throughput; miniaturization; phenotypic screening