Summary: This invention provides a high-throughput, cost-effective, isoform-selective, human-based HDAC activity assay useful for understanding the activity of the 11 different HDAC isoforms present in humans. HDAC (Histone deacetylase) proteins are key regulators in gene expression and HDAC inhibitors represent an important new drug discovery target for cancer and a growing number of other diseases. The WSU technology uses immunoprecipitation of individual HDAC proteins from human cell lysates to develop the first enzyme linked immunosorbent (ELISA) based assay for monitoring HDAC activity.
While numerous companies such as Promega, Millipore and Enzo Biosciences make HDAC activity assays for basic science, screening and drug development research, the high cost of purified HDAC isoforms expressed from baculovirus systems limits high throughput use. The WSU invention significantly reduces the cost and uses human cell-derived HDAC isoforms. Note the use of human derived proteins is a significant advantage given inhibitor screens using baculovirus proteins show significant disparities in measured IC50 values compared to mammalian derived proteins. The use of miniature well plates helps avoid nonspecific interactions between isoforms and greatly reduce the cost per test. Because the assay overcomes the intrinsic limitations of current screening technology, the methods are expected to significantly accelerate identification of selective HDAC inhibitors.
Benefits:
• Assay for isoform selective HDAC inhibitors
• Uses human cell derived HDAC isoforms (not baclovirus expressed)
• Significantly lower costs per test
• Works on high throughput surface plates
• Compatible with numerous detection labels
Stage of Development:
• Patent Pending
Tested several 96 well plate prototypes using a sensitive luminescent signal (HDAC-Glo) to determine the isoform selective activity after incubation with a small library of inhibitors. Sample results shown below
