Category: Assay Development and Screening
Ectonucleotidases are plasma membrane-bound enzymes with externally oriented active sites that metabolize nucleotides to nucleosides and are crucial for maintaining immune homeostasis. The ectonucleoside triphosphate diphosphohydrolase-1, also known as CD39, ENTPD1, or NTPDase1 hydrolyzes ATP and ADP to AMP. AMP can further be processed to adenosine leading to a significant impact on many disease states. Recent studies have shown a key role for adenosine in immunosuppression in the tumor microenvironment, and ectonucleotidases are emerging as promising immuno-oncology targets. As the only HTS method capable of direct detection of nucleotides, the Transcreener platform is uniquely suited for measuring ectonucleotidase activity with the high sensitivity and low levels of interference required for a successful HTS campaign. The homogenous assays use a far-red fluorescence polarization (FP) or TR-FRET readout and they can be broadly applicable to ectonucleotidases. We developed a simple biochemical assay for measuring CD39 activity using the Transcreener AMP2 Assay. The assay provides robust detection of AMP production (Z’ > 0.6) with sub-nanomolar amounts of CD39. Initial pilot screens have demonstrated robust assay performance (Z’ = 0.6 – 0.7) and IC50s determined for tool compounds of CD39 were consistent with published values. The availability of HTS-compatible enzyme assay methods will accelerate the discovery of inhibitors for CD39 and related ectonucleotidases that play a role in tumor immunity and other diseases impacted by adenosine signaling.
Justin Brink– Director of Marketing, BellBrook Labs, Madison, Wisconsin
Director of Marketing
With years of assay development experience, I am currently working to find applications and technology to advance drug discovery. My goal is to discover areas of research where we can leverage current technology platforms to enhance the process of finding new treatments for disease. In my current role with BellBrook Labs, we work every day to accelerate drug discovery by providing HTS solutions to our academic and biotech collaborators. My hope is that this work will provide lasting benefit, and treatments for a multitude of ailments for people worldwide.