Evaluation of Disulfide Bond Scrambling in mAb peptide Mapping

May 20, 2026

Peptide mapping under non‑reducing conditions enables direct characterization of native disulfide connectivity in proteins such as monoclonal antibodies, but method‑induced disulfide scrambling remains a significant challenge. Reactive thiol intermediates formed during sample preparation can generate artificial disulfide rearrangements, obscuring true cysteine connectivity and complicating structural confirmation and comparability assessments. 

This poster presents work from RIC Group, featuring an automated peptide mapping workflow in Genedata Expressionist^®, enabling detection, quantification, and differentiation of artificial and real disulfide scrambling events from MS/MS data. You will learn how automated data processing reduces labor‑intensive manual handling, supports accurate identification of disulfide‑bonded species, and enables reliable quantification of scrambling under different stress conditions. It also demonstrates how targeted analytical strategies and traceable workflows improve confidence in disulfide bond characterization while minimizing method‑induced artifacts. Designed for scientists working with peptide mapping and structural characterization of biotherapeutics.