Abstract
Covalent chemical probes are able to display incomparable potency and selectivity while targeting challenging binding sites and non-traditional protein targets. The development of such covalent probes entails additional requirements. On the one hand, covalent probes require careful accounting for their reactivity, the evaluation of formation of the covalent bond and kinetic measurement of activity. On the other hand, they also allow relatively simple target cellular engagement assessment and an elaborate view of cellular off-targets. This chapter describes common methods, examples and best practices for the discovery, optimization and evaluation of covalent chemical probes and suggests general criteria to strive for in their development.
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