The goal of assay development for interaction-based screening methods is finding the conditions that ensure the best signal to noise ratio and assay stability over time. The search for these optimal assay conditions is a long and costly process due to the large number of variables that have to be tested. Access to a systematic approach that examines a large number of variables at once can reduce the complexity of assay development for interaction-based screening methods — which translates into a faster and less costly process. Here we present a novel, fully automated and systematic plate-based approach to the assay development of a biophysical method for interaction based screening utilizing NanoTemper’s Buffer Exploration Kit. With this method, we tested 96 different buffer conditions for the interaction-based screening of histone methyltransferase G9a in less than 5 hours. Amongst the conditions we tried were various buffer systems, sodium chloride (NaCl) concentrations, addition of Tween®20 and Pluronic®F-127, as well as a variety of additives. One significant discovery was that the signal to noise ratio of the assay increased by at least two-fold in the presence of divalent cations.
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