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Published on: August 10, 2017
Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors
Geetha Padige1, Ahmed T Negmeldin1, Mary Kay H Pflum2
1Department of Chemistry, Wayne State University, Detroit, MI, USA.
Abstract:
Histone deacetylase (HDAC) proteins are promising targets for cancer treatment, with several HDAC inhibitors used clinically as anticancer drugs. Most HDAC inhibitors nonspecifically interact with all or many of the 11 HDAC isoforms. Isoform-selective HDAC inhibitors would be useful tools to dissect the individual functions of HDAC proteins in cancer formation, in addition to potentially displaying effective anticancer properties. We report here a robust HDAC activity assay for screening selective HDAC inhibitors, which is inspired by the traditional enzyme-linked immunosorbent assay (ELISA). The key feature of the ELISA-based HDAC activity assay is use of mammalian cell-derived HDAC isoforms instead of recombinant proteins. Importantly, the assay was validated with several known HDAC inhibitors. The ELISA-based HDAC activity assay will facilitate the characterization of isoform-selective HDAC inhibitors against mammalian cell-derived HDAC proteins, which will enhance HDAC-centered cancer research and provide a foundation for anticancer drug development.
Insights
A new enzyme-linked immunosorbent assay (ELISA) enables screening of histone deacetylase (HDAC) inhibitors. This assay uses cell-derived HDACs, advancing cancer research and drug development for isoform-selective HDAC inhibitors.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Histone deacetylase (HDAC) proteins are crucial targets in cancer therapy.
- Current HDAC inhibitors lack isoform specificity, limiting their therapeutic potential and research applications.
- Developing isoform-selective inhibitors requires robust assays for screening.
Purpose of the Study:
- To develop a novel, robust assay for screening isoform-selective HDAC inhibitors.
- To utilize mammalian cell-derived HDAC isoforms for more biologically relevant screening.
- To facilitate the discovery of targeted HDAC-centered cancer therapies.
Main Methods:
- An enzyme-linked immunosorbent assay (ELISA)-based method was developed to measure HDAC activity.
- The assay utilizes HDAC isoforms derived from mammalian cells, not recombinant proteins.
- The assay's performance was validated using known HDAC inhibitors.
Main Results:
- A reliable ELISA-based HDAC activity assay was successfully established.
- The assay demonstrated effectiveness in screening for selective HDAC inhibitors.
- Validation with known inhibitors confirmed the assay's utility.
Conclusions:
- The developed ELISA-based HDAC activity assay is a valuable tool for identifying isoform-selective HDAC inhibitors.
- This assay will accelerate research into the specific functions of HDACs in cancer.
- It provides a foundation for developing novel, targeted anticancer drugs.
