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An Automated Bioluminescent Assay for Measuring Phosphodiesterase Activity Scientific...

An Automated Bioluminescent Assay for Measuring Phosphodiesterase Activity Scientific Poster

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Abstract

Tracy Worzella1, Kevin Hsiao1, Said Goueli1, Harris Grevelis2, and Patricia Ahrweiler2
1Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711;
2CyBio, Inc., 500 West Cummings Park Suite 1800, Woburn, MA 01801
Phosphodiesterases (PDE) represent a family of proteins that play important roles in signal transduction and signal amplification by regulating intracellular levels of second messengers cAMP and cGMP. PDE involvement in immune and inflammatory responses, hypertension and neuronal activity makes them an attractive therapeutic target. The 'drug-ability' of this target is exemplified with recent approvals of PDE inhibitors for erectile dysfunction. To aid in the discovery of new PDE inhibitors, we have developed an automated application of a bioluminescent phosphodiesterase assay, the PDE-Glo™ Assay, using the CyBio CyBi®-Well 384/1536 liquid handling system. The PDE-Glo™ Assay is used to measure cyclic nucleotide phosphodiesterase activity from purified sources. The homogeneous, add-mix-measure format of this assay makes it ideal for performing assays in high density plate formats. Here we demonstrate the robustness of the assay in low volume 384- and 1536-well formats using the CyBi®- Well for assay setup. We show the utility of the assay for detecting PDE inhibitors, both in a small-scale screen and in dose response studies. The PDE-Glo™ Assay is optimized to work equally well with both cAMP- and cGMPdependent phosphodiesterases. Here we show the utility of the assay in measuring cAMP-dependent phosphodiesterase activity using purified PDE bovine brain.

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  • Part# PS007
  • Printed in USA.

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