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Related Experiment Video

Updated: Jun 22, 2026

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
08:10

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

Enzyme assay design for high-throughput screening.

Kevin P Williams1, John E Scott

  • 1Department of Pharmaceutical Sciences and BRITE, North Carolina Central University, Durham, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 25, 2009
PubMed
Summary
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This chapter outlines standardized methods for developing and validating enzyme assays, crucial for drug discovery. It provides practical solutions for common challenges in high-throughput screening (HTS) of enzyme targets.

Area of Science:

  • Biochemistry and Pharmacology
  • Drug Discovery and Development

Background:

  • Enzymes are key targets in pharmaceutical research, with high-throughput screening (HTS) being the primary method for identifying active compounds.
  • Established guidelines exist for enzyme assay development and validation, benefiting from extensive academic and industrial experience.

Purpose of the Study:

  • To generalize and present approaches for developing, validating, and troubleshooting enzyme assays applicable in both industrial and academic settings.
  • To provide practical solutions and examples for common assay development challenges.

Main Methods:

  • Generalization of assay development and validation approaches for enzyme screens.
  • Illustrative examples covering diverse enzyme classes: kinases, proteases, transferases, and phosphatases.

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High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
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A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

Published on: August 17, 2019

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Last Updated: Jun 22, 2026

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
08:10

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

Published on: March 3, 2015

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
10:28

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

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  • Discussion of practical strategies for handling low-purity targets, compound interference, and activator identification.

    • Main Results:

    • A generalized framework for enzymatic assay development and validation is presented.
    • Real-world examples demonstrate solutions for common issues encountered in enzyme screening.
    • Guidance on assay acceptance criteria and potential pitfalls is provided.

    Conclusions:

    • The chapter offers a comprehensive guide to developing robust enzymatic assays for HTS.
    • Standardized approaches and practical troubleshooting tips enhance the efficiency of drug discovery efforts.
    • The presented methodologies are broadly applicable across various enzyme classes and research settings.