Hedgehog Pathway Antagonist 5E1 Binds Hedgehog at the Pseudo-Active Site

Maun, H.R., et al., 285(34), 26570-26580, J Biological Chemistry, 2010

A murine/human chimeric 5E1 monoclonal antibody (ch5E1) against the hedhehog pathway was engineered. Binding kinetics and affinities of ch5E1 Fab for various hedgehog ligands were measured using the Octet RED platform. A number of kinetic binding experiments were reproted using biotinylated Sonic hedgehog, biotinylated ch5E1 Fab, or biotinylated hedgehog ligand immobilized on Streptavidin biosensors. Kinetics experiements were confirmed in reverse orientations. In the presence of calcium and zinc ions, ch5E1 binding affinity for Sonic hedgehog increased 10-20-fold, primarily because of a decrease in the dissociation rate.

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