title: An Automated Microscale Thermophoresis Screening Approach for Fragment-Based Lead Discovery.
abstract: Fragment-based lead discovery has proved to be an effective alternative to high-throughput screenings in identifying chemical matter that can be developed into robust lead compounds. The search for optimal combinations of biophysical techniques that can correctly and efficiently identify and quantify binding can be challenging due to the physicochemical properties of fragments. In order to minimize the time and costs of screening, optimal combinations of biophysical techniques with maximal information content, sensitivity, and robustness are needed. Here we describe an approach utilizing automated microscale thermophoresis (MST) affinity screening to identify fragments active against MEK1 kinase. MST identified multiple hits that were confirmed by X-ray crystallography but not detected by orthogonal methods. Furthermore, MST also provided information about ligand-induced aggregation and protein denaturation. The technique delivered a large number of binders while reducing experimentation time and sample consumption, demonstrating the potential of MST to execute and maximize the efficacy of fragment screening campaigns.
authors.affiliation: NanoTemper Technologies GmbH, Munich, Germany.
authors.forename: Pawel
authors.initials: P
authors.lastname: Linke
authors.affiliation: Sanofi R&D, Structure-Design-Informatics, Vitry sur Seine, France.
authors.forename: Kwame
authors.initials: K
authors.lastname: Amaning
authors.affiliation: NanoTemper Technologies GmbH, Munich, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Maschberger
authors.affiliation: Sanofi R&D, Structure-Design-Informatics, Vitry sur Seine, France.
authors.forename: Francois
authors.initials: F
authors.lastname: Vallee
authors.affiliation: Sanofi R&D, Structure-Design-Informatics, Vitry sur Seine, France.
authors.forename: Valerie
authors.initials: V
authors.lastname: Steier
authors.affiliation: NanoTemper Technologies GmbH, Munich, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Baaske
authors.affiliation: NanoTemper Technologies GmbH, Munich, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Duhr
authors.affiliation: NanoTemper Technologies GmbH, Munich, Germany dennis.breitsprecher@nanotemper.de.
authors.forename: Dennis
authors.initials: D
authors.lastname: Breitsprecher
authors.affiliation: Sanofi R&D, Structure-Design-Informatics, Vitry sur Seine, France.
authors.forename: Alexey
authors.initials: A
authors.lastname: Rak
mesh_terms: D018360:Crystallography, X-Ray; D004058:Diffusion; D055808:Drug Discovery; D015870:Gene Expression; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008024:Ligands; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors / Q000737:chemistry*; D008958:Models, Molecular; D011485:Protein Binding; D011489:Protein Denaturation; D047428:Protein Kinase Inhibitors / Q000737:chemistry*; D054852:Small Molecule Libraries / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D013696:Temperature
chemical_list: D008024:Ligands; D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; D048369:MAP Kinase Kinase 1; C482125:MAP2K1 protein, human
keywords: binding affinity; biophysical screening; drug discovery; protein aggregation; surface plasmon resonance