Intact antibody structural characterization using H/D exchange and top-down electron transfer dissociation on an Orbitrap

摘要

Top-down HDX was applied to an 150 kDa intact protein. Individual domain-level deuteration information was obtained for 6 of the 12 IgG domains on Herceptin, which included the antigen binding sites. Protein binding was localized to the VL domain for Herceptin. Back-exchange was reduced to 2% by lowering the HPLC temperature to -20°C.