TECHNICAL NOTE
Fast-track your hit screening assay development
with the Buffer Exploration Kit
Abstract
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.
Introduction
Hit screening for drug discovery can be done using a variety of biochemical and
biophysical techniques that characterize molecular interactions and ultimately quantitate
binding affinities. A fundamental step during assay optimization for any of these methods
Alina Neumann, Amit J. Gupta and Johannes Rieger
NanoTemper Technologies GmbH, Munich, Germany
