UNIT 9.4 Overview of High-Throughput Screening

  1. Michael Entzeroth,
  2. Horst Flotow,
  3. Peter Condron

Published Online: 1 MAR 2009

DOI: 10.1002/0471141755.ph0904s44

Current Protocols in Pharmacology

Current Protocols in Pharmacology

How to Cite

Entzeroth, M., Flotow, H. and Condron, P. 2009. Overview of High-Throughput Screening. Current Protocols in Pharmacology. 44:9.4:9.4.1–9.4.27.

Author Information

  1. Experimental Therapeutics Centre, Agency for Science, Technology, and Research (A*STAR), Singapore

Publication History

  1. Published Online: 1 MAR 2009
  2. Published Print: MAR 2009


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.


  • high-throughput screening;
  • high-content assays;
  • functional assays;
  • data management;
  • hit identification;
  • lead optimization