Analytical methods for protein kinase and inhibitor screening including kinetic evaluation

Protein kinases promote cell processes such as cell growth and proliferation similarly in tumor cells and healthy cells. Pharmaceutical research aims at developing small molecules that inhibit tumor growth binding in the ATP-binding pocket of specific protein kinases. To understand the specificity of the inhibitor, thermodynamic and kinetic data of the kinase-inhibitor binding process are important. A broad range of analytical methods may investigate the interactions between protein kinases and inhibitors. Today, however, there is no comprehensive strategy to provide information on both specificity screening and binding kinetics. The purpose of this review is to summarize analytical methods used not only to monitor the protein kinase inhibition, but also to provide kinetic and thermodynamic data on the basis of screening applications. A focus is set on optical biosensing as the most promising techniques. We discuss, that intelligent combinations of methods can provide comprehensive information on protein kinases interacting with inhibitors. • Mainly biosensors provide binding affinity and kinetics at low costs and sample amounts. • Comprehensive analysis is reached with identification by mass spectrometry. • The biosensor's surface chemistry to beads allows kinase capturing. • The selectivity of the beads can be tuned via the choice of (selective) inhibitors.