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Drug Discovery at the Speed of Sound
High-Throughput Affinity Screening using Monolith NT.Automated
Dr. Christian Kleusch | Technical Sales Specialist
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M o n o l i t hN T . 1 1 5
M o n o l i t hN T . A u t o m a t e d
S o l u t i o n s f o r f u l l a u t o m a t i o n
2 0 1 2 2 0 1 42 0 1 4 2 0 1 61 8 5 6 / 1 8 7 9
Carl Ludwig
Charles Soret
Foundation: LMU Munich
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Fast thermo-optical particlecharacterisationPCT Patent 2008/061706
Red Dot Design Award
German Innovation Award
P r o m e t h e u sN T . 4 8
German FoundersPrize
2 0 1 7
Growth Champion
Screening Campaigns with the Monolith NT.Automated
Introduction to MicroScale Thermophoresis Screenings
• Short introduction to biophysical method Microscale Thermophoresis (MST)
• Single Point Screening
• Affinity Screening
• Case studies: compound screens against MEK1 (Sanofi), p38α (NanoTemper), HSP90 (Vernalis)-> some facts and figures
MicroScale Thermophoresis
DNA
Duhr and Braun, PNAS, 2006Duhr and Braun, PRL, 2006
Theory of thermophoresis: contributing factors
c1 / c0 = exp (-ST ΔT)
Size Charge2Hydration Shell
Duhr and Braun, PNAS, 2006Duhr and Braun, PRL, 2006
Optical MicroScale Thermophoresis
MicroScale Thermophoresis: Physical Processes
Thermophoresis ~ 30 sec
23°C
20°C
1K/10 µm corresponds to 1000K/cm
IR laser induced temperature Ffeld
Jerabek-Willemsen et al, ADDT, 2011
Sample heating < 1 sec
MST Technology
MST signal
MST binding curve
unbound
bound
F0 F1
Fit
Kd
Two Screening Setups
Primary screening: Single Point
• Primary hit selection (binder vs non-
binder)
• Single concentration of each compound
Secondary screening: Affinity
• Compound ranking
• Serial dilution of each compound
• 8 points per compound ► 12 Kds
per run w run time 80s/KD
Automated Screening utilizing MicroscaleThermophoresis
MST for Single Dose Screenings
• Detect binding within seconds
• Identify false positives quickly
Fully automated sample preparation, liquid handling and MST detection
using a Monolith NT.Automated in conjunction with a Hamilton Starlet
0. Ligand stock in 384 well plate
Hit Selection
Signal amplitude is independent of
binding affinity!
Affinity ScreeningMST for Kd ranking
• Automized Kd ranking
• Identify false positives quickly
Fully automated sample preparation, liquid handling and MST detection
using a Monolith NT.Automated in conjunction with a Hamilton Starlet
0. Ligand stock in 384 well plate
MO.Screening Analysis Software
Automated Screening of p38a Kinase Inhibitors
1. Hit identification 2. Affinity Characterization
Bartoschik et al., NanoTemper Technologies
Applied Biophysics in Drug Discovery
MST fragment screen against Hsp90 N-terminal domain
In collaboration with
Automated screening of fragments against MEK1 kinase
• Find unique hits and more lead candidates
• Easily identify false positives to streamline the discovery process
One of seven MST-unique hitsbound to MEK1
Linke et al., NanoTemper Technologies & SanofiJournal of Biomolecular Screening, 2014
SPR rank #1
SPR rank #2
Identification of false-positives
► MST identifies false positives through fluorescence loss due to protein denaturation.
► MST identifies false positives through aberrant MST traces.
The added value of automated MST • Get results quickly, from challenging targets
Measure 12 Kds in 16 minutes. Use < 100 pg target to screen > 200 compounds.
• Benefit from close-to-native conditions Immobilization-free interactions in all buffers and bioliquids (cell lysate, serum).
• Do your research efficiently
Enjoy perfect ease-of-use and get rid of maintenance downtime. Fully automated sample handling. Unique quality controls intrinsic to MST.
• Generate results right from the start
Groningen Drug Design group uses MST since years already. Publications, PoC, Assay-Development – all in their hands already.
Thank you for your attention.christian.kleusch@nanotempertech.com | support@nanotempertech.com
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