TR-FRET Based Transcreener AMP²/GMP² Assay for Screening Ligases, Synthetases and Phosphodiesterases

The exploration of more extensive drug target space by pharmaceutical and academic labs is creating a need for HTS compatible methods to detect diverse enzymes. The Transcreener AMP²/GMP² Assay is simple, single step, homogenous, amenable to HTS and enables the detection of any AMP/GMP producing enzyme while using any precursor substrate. Importantly, the assay relies on direct immunodetection of AMP and GMP without using coupling enzymes, which eliminates the need for counter-screening. The readout options have been extended to include far-red, time-resolved Förster-resonance-energy-transfer (TR-FRET) in addition to the original fluorescence polarization based assay. This poster describes the development, optimization and validation of the Transcreener AMP²/GMP² TR-FRET Assay, including detection of different classes of AMP/GMP-producing enzymes.