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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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

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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

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German FoundersPrize

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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