APPLICATION NOTE
1
©2019 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved.
Fast molecular interaction screening of epigenetic
gene regulator G9a with fragments from a large
chemical space
Anne Neumann
1
, Alina Neumann
3
, Tanja Bartoschik
3
, Johannes Rieger
3
, Amit J. Gupta
3
,
Peter Cherepanov
1
and Matthew J. Fuchter
2
1
The Francis Crick Institute, 1 Midland Road, NW1 1AT, London, United Kingdom
2
Imperial College London, Department of Chemistry, Molecular Sciences Research Hub, Wood Lane, W12 0BZ, London, United Kingdom
3
NanoTemper Technologies GmbH, Flößergasse 4, 81369 München, Germany
Abstract
One of the latest trends in drug discovery screening shows a rise in the number of
explorative targets. In order to evaluate these targets, the industry needs screening
technologies that offer flexibility in characterizing binding events between more
heterologous targets and ligands from large or new chemical spaces. Dianthus
NT.23PicoDuo swi ly detects and quantifies molecular interactions in solution,
independent of the molecular mass of the two interacting molecules, making it
a perfect match for fragment-based screening with compounds representing a
large chemical space. In this application note, we show data from single-dose
screening of a fragment library with a broad molecular mass and complexity
range against the target molecule histone methyltransferase (HMT) G9a, followed
by affinity constant determination (K
d
) for a selection of hit molecules. At the end
of the study, we screened 5,784 data points including controls in less than 8 hours
of instrument time, using less than 20 µg of target protein. Affinities were in the
high micromolar range as expected from fragments, and in good agreement with
published values for the positive control.