Characterization Of An Antibody-Drug-Conjugate (ADC) Using Electron Activated Dissociation (EAD)

By Zoe Zhang, Takashi Baba, Pavel Ryumin, Bill Lloyd, Jason Causon, and Kerstin Pohl

With the development in protein engineering, antibodies and their related derivatives become the fastest growing class of therapeutics. Antibody-drug-conjugates are one of those new modalities. ADCs are often composed of a 150 kDa monoclonal antibody (mAb) covalently coupled with cytotoxic payloads, or other types of drugs, through synthetic linkers. 

A new, highly reproducible fragmentation type based on EAD was used to analyze the conjugated peptides from a commercial ADC. The data were acquired with an untargeted 10 Hz rapid data-dependent acquisition (DDA) method and interpreted with Protein Metrics Inc. software. With this workflow, regular and advanced characterization leveraging EAD-based fragmentation is achievable in one injection, enabling a streamlined characterization accessible to every user-level.