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A P P L I C A T I O N N O T E
L I S T O F A U T H O R S
Ivana Jaser, Vanessa Hösl, Jan Schnatwinkel, Nathan B. P. Adams
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NanoTemper Technologies, Tölzer Straße 1, Munich, Germany
Contact: nathan.adams@nanotempertech.com
A B S T R A C T
Here, we introduce a new ultra-high throughput biophysical screening instrument, the Dianthus™
uHTS and provide an example screening data set exploring small-molecule inhibitors targeting MEK1,
a key kinase in the MAPK/ERK pathway. The Dianthus uHTS platform enables ultra-high-throughput
biophysical screening (approx. 250 000 data points per day) by detecting ligand interactions using
Spectral Shi technology. In this study, we screened 5120 different compounds against MEK1,
identifying 62 high priority hits by single dose screening and further 81 hits of a lower priority. The
dose-response analysis confirmed 28 compounds with K
d
values below 10 µM, including five with
affinities sub 100 nM. Orthogonal validation on high priority compounds using thermal shi and ADP
Glo assays further refined the hit list to 6 validated binders which also inhibit ATPase activity. These
results demonstrate the efficiency of Dianthus uHTS for rapid hit identification with minimal sample
consumption, supporting its application in early-stage drug discovery.
Ultra-high-throughput biophysical
screening of MEK1 using the
Dianthus uHTS platform
K E Y W O R D S
Dianthus uHTS
Biophysical screening
Dose-response
Protein kinase
Hit identification
