An Automated Microscale Thermophoresis Screening Approach for Fragment-Based Lead Discovery
| Autor: | François Vallée, Philipp Baaske, Melanie Maschberger, Valerie Steier, Kwame Amaning, Pawel Linke, Stefan Duhr, Alexey Rak, Dennis Breitsprecher |
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| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular Protein Denaturation Fragment-based lead discovery MAP Kinase Kinase 1 Gene Expression Nanotechnology Computational biology Crystallography X-Ray Ligands Biochemistry drug discovery protein aggregation Analytical Chemistry Diffusion Small Molecule Libraries biophysical screening Fragment (logic) Robustness (computer science) binding affinity High-Throughput Screening Assays Humans Protein Kinase Inhibitors Chemistry Drug discovery Microscale thermophoresis Temperature Surface Plasmon Resonance Molecular Medicine Technical Notes Protein Binding Biotechnology |
| Zdroj: | Journal of Biomolecular Screening |
| ISSN: | 2472-5552 |
| DOI: | 10.1177/1087057115618347 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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