Related Experiment Video
Updated: Jun 23, 2026
Assays for Validating Histone Acetyltransferase Inhibitors
Published on: August 6, 2020
Non-isotopic dual parameter competition assay suitable for high-throughput screening of histone deacetylases
Daniel Riester1, Christian Hildmann, Patricia Haus
1University of Göttingen, Department of Molekular Genetics and Praeparative Molekular Biology, Grisebachstr. 8, 37077 Goettingen, Germany.
Abstract:
Histone deacetylases reside among the most important and novel target classes in oncology. Selective lead structures are intensively developed to improve efficacy and reduce adverse effects. The common assays used so far to identify new lead structures suffer from many false positive hits due to auto-fluorescence of compounds or triggering undesired signal transduction pathways. These drawbacks are eliminated by the dual parameter competition assay reported in this study. The assay involves a new fluorescent inhibitor probe that shows an increase in both, fluorescence anisotropy and fluorescence lifetime upon binding to the enzyme. The assay is well suited for high-throughput screening.
Insights
This study introduces a novel dual parameter competition assay for identifying cancer drug leads. The assay uses a fluorescent probe to overcome false positives common in current methods, enabling more reliable high-throughput screening.
Area of Science:
- Oncology
- Biochemistry
- Assay Development
Background:
- Histone deacetylases (HDACs) are key targets in cancer therapy.
- Current assays for HDAC inhibitors suffer from false positives due to compound auto-fluorescence and pathway activation.
- Development of selective HDAC inhibitors is crucial for improved cancer treatment efficacy and reduced side effects.
Purpose of the Study:
- To develop a novel, reliable assay for identifying selective histone deacetylase inhibitors.
- To overcome limitations of existing assays, such as false positives.
Main Methods:
- A dual parameter competition assay was designed.
- The assay utilizes a new fluorescent inhibitor probe.
- The probe's binding to the enzyme is detected by changes in fluorescence anisotropy and fluorescence lifetime.
Main Results:
- The developed assay effectively eliminates false positive hits common in conventional methods.
- The assay demonstrates increased fluorescence anisotropy and fluorescence lifetime upon inhibitor binding.
- The assay is suitable for high-throughput screening of potential HDAC inhibitors.
Conclusions:
- The novel dual parameter competition assay provides a robust method for identifying selective histone deacetylase inhibitors.
- This assay overcomes significant drawbacks of existing screening methods, improving reliability.
- The assay is well-suited for high-throughput screening in oncology drug discovery.
