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Updated: May 30, 2026

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
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A bioluminogenic HDAC activity assay: validation and screening.

Francoise Halley1, Jeanette Reinshagen, Bernhard Ellinger

  • 1European ScreeningPort GmbH, Hamburg, Germany.

Journal of Biomolecular Screening
|August 12, 2011
PubMed
Summary
This summary is machine-generated.

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New bioluminogenic assays for histone deacetylase (HDAC) enzymes offer reduced compound interference for drug discovery. These validated assays are suitable for high-throughput screening of potential therapeutics targeting HDACs in various diseases.

Area of Science:

  • Biochemistry
  • Enzymology
  • Drug Discovery

Background:

  • Histone deacetylase (HDAC) enzymes regulate protein acetylation, and their dysregulation is linked to numerous diseases.
  • Developing targeted drugs for HDACs is crucial for therapeutic intervention.
  • Existing fluorescence-based assays can suffer from compound interference, limiting their utility.

Purpose of the Study:

  • To evaluate homogeneous, single-addition, bioluminogenic HDAC enzyme activity assays.
  • To assess the performance of these assays for high-throughput screening (HTS) applications.
  • To compare bioluminogenic assays with traditional fluorescence-based formats regarding compound interference.

Main Methods:

  • Assessed key assay parameters: sensitivity, scalability, reproducibility, signal stability, Z-factor, DMSO tolerance, and inhibitor response.

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  • Evaluated assays against HDAC-1, HDAC-3/NcoR2, HDAC-6, and SIRT-1 enzymes.
  • Miniaturized assays to 10 µL volume and validated using FDA-approved drugs and a natural products library.

    • Main Results:

    • Bioluminogenic assays demonstrated reduced interference from test compounds compared to fluorescence assays.
    • Assays were successfully miniaturized and showed high-throughput screening compatibility.
    • Screening of FDA drugs and natural products confirmed assay performance in various assay formats.

    Conclusions:

    • Homogeneous, bioluminogenic HDAC assays are robust, sensitive, and suitable for HTS.
    • These assays offer an advantage over fluorescence formats due to lower compound interference.
    • Validated assays provide a valuable tool for discovering novel HDAC-targeting therapeutics.