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Studying the binding affinity of different ERK2 binding partners with MicroScale Thermophoresis Clemens Entzian, Lukas Kniep, Dr. Thomas Schubert, 2bind GmbH Antibody 1B3B9 (Merck) Molecular Size ERK2 is an important player in multiple biochemical signaling pathways and is involved in different cancer types. This highly relevant pharmaceutical target represents an optimal model system to demonstrate the capabilities of our MicroScale Thermophoresis (MST) platform. fraction bound 1 0.8 0.6 150 kDa 0.4 0.2 0 0.1 1 10 100 1000 104 concentration titrant/nM Kd = 137.9 + 23.3 nM Aptamer C5.71 The results (figure to the right) demonstrate that our MST platform is perfectly suited to study molecular interactions independent of the size and mass of the ligand. Therefore, MicroScale Thermophoresis is the optimal tool to characterize drug-target, antibody-antigen or aptamer-target interactions in terms of basic binding parameters. 0.4 3 kDa 0.2 0 0.1 1 10 100 1000 104 Erk Activation Inhibitor Peptide I (Merck) 1.2 fraction bound • ERK2 Inhibitor I (Merck) • ERK2 Activation Inhibitor Peptide I (Merck) • Aptamer C5.71 (Mayer lab, Bonn) • Antibody 1B3B9 (Merck) 0.6 concentration titrant/nM Kd = 171.1 + 36.0 nM 0.8 1 kDa 0.4 0 10 100 1000 104 105 106 concentration titrant/nM Kd = 4854.2 + 1686 nM ERK2 Inhibitor I (Merck) 1.2 fraction bound In this example study, the binding affinity of ERK2 (MAPK1, mitogen-activated protein kinase 1) towards the following four different binding partners (size range from 0.3 kDa to 150 kDa) was studied by MST: fraction bound 1 0.8 0.8 0.4 0.3 kDa 0 1 10 100 1000 104 105 106 concentration titrant/nM Kd = 2025 + 924 nM www.2bind.de - Service provider for biophysical analyses - from stability to affinity