Supplementary Figure 2: Protease cleavage of coiled-coil masked antibodies.
From: A coiled-coil masking domain for selective activation of therapeutic antibodies
a) Representative characterization of coiled-coil masked antibodies and MMP activation by LC-MS. Shown is the deconvoluted spectra of CC2B-IPVSLRSG-rituximab heavy and light chains (top) as well as MMP-2-cleaved CC-IPVSLRSG-rituximab heavy and light chains (bottom) following reduced reverse-phase LC-MS. b) Representative data for the cleavage of CC3-IPVSLRSG-hBU12 with a panel of human (h) and mouse (m) MMPs. Activated MMP (6 Enzyme Units, EU, amount of enzyme to process 1 μg substrate/minute) was incubated with antibody (10 μg) at for 20 min at 37 °C. Reactions were stopped by addition of 25 mM EDTA and samples were analyzed using the method described in (A). The percent cleaved heavy or light chain was calculated using peak intensities from deconvoluted mass spectra. Two independent experiments were performed with similar results.
