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Practical guidelines for the AMI-Isonitrile ligation
Ian Warm1, Athanasios Markos2, Helma Wennemers1
1Laboratory of Organic Chemistry, D-CHAB, ETH Zürich, Zürich, Switzerland.
None:
The azomethine imine (AMI)-isonitrile ligation is a fast and robust bioorthogonal reaction with unique pH-dependent kinetics. In this chapter, we provide practical guidelines for the synthesis of isonitrile reagents, functionalizable AMIs and AMI-fluorophore conjugates. We describe detailed protocols for the installation of isonitrile handles onto proteins via thiol-maleimide or amine-reactive ester chemistries and onto live cells via reactive ester or antibody-antigen-mediated approaches. Importantly, we provide detailed conditions for the functionalization of the introduced isonitriles by the AMI-isonitrile ligation.
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