Overview of High‐Throughput Screening

Michael Entzeroth1, Horst Flotow1, Peter Condron1

1 Experimental Therapeutics Centre, Agency for Science, Technology, and Research (A*STAR), Singapore
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 9.4
DOI:  10.1002/0471141755.ph0904s44
Online Posting Date:  March, 2009
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Abstract

High‐throughput screening (HTS) is a key process used in drug discovery to identify hits from compound libraries that may become leads for medicinal chemistry optimization. This updated overview discusses the utilization of compound libraries, compounds derived from combinatorial and parallel synthesis campaigns and natural product sources; creation of mother and daughter plates; and compound storage, handling, and bar coding in HTS. The unit also presents an overview of established and emerging assay technologies (i.e., time‐resolved fluorescence, fluorescence polarization, fluorescence‐correlation spectroscopy, functional whole cell assays, and high‐content assays) and their integration in automation hardware and IT systems. This revised unit provides updated descriptions of state‐of‐the‐art instrumentation and technologies in this rapidly changing environment. The section on assay methodologies now also covers enzyme complementation assays and methods for high‐throughput screening of ion channel activities. Finally, a section on criteria for assay robustness is included discussing the Z′‐factor, which is now a widely accepted criterion for evaluation and validation of high throughput screening assays. Curr. Protoc. Pharmacol. 44:9.4.1‐9.4.27. © 2009 by John Wiley & Sons, Inc.

Keywords: high‐throughput screening; high‐content assays; functional assays; data management; hit identification; lead optimization

     
 
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Table of Contents

  • Introduction
  • High‐Throughput Screening
  • Precedents
  • Compound Libraries
  • Combinatorial/Parallel Synthesis Chemistry
  • Natural Products
  • Compound Storage and Handling
  • Assay Formats
  • Mother and Daughter Plates
  • Barcodes
  • Assay Design
  • Classical Receptor and Enzyme Assays
  • Homogeneous Radioactive Bioassays
  • Fluorescence
  • Luminescence
  • Reporter‐Gene Assays
  • Cell‐Based Assays for GPCR and Ion Channel Targets
  • High‐Content Assays and Screening
  • Assay Robustness
  • HTS Informatics
  • Robotic Versus Automated Workstations
  • Hit‐to‐Lead Process
  • Is it Worth the Effort?
  • Outlook and Further Directions
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Literature Cited

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