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Discovery of a Photoinduced Histidine-Histidine Cross-Link in an IgG4 Antibody

Thomas Powell1, Michael J Knight1, John O'Hara1

  • 1Biomolecular Formulation and Characterization Sciences, UCB Group, Slough, SL3WE, United Kingdom.

Journal of the American Society for Mass Spectrometry
|May 12, 2020
PubMed

A novel histidine-histidine (His-His) photooxidative cross-link has been identified in an IgG4 antibody. It was formed between the side chain of a histidine residue of the antibody and histidine from the formulation buffer. The structure of the cross-link was elucidated using high performance liquid chromatography (HPLC) hyphenated to tandem mass spectrometry (MS/MS) with higher energy collisional dissociation (HCD). The cross-link was found in multiple conformations, as the location of the oxygen varied. Furthermore, the extent of cross-link formation was shown to correlate with the amount of light the antibody was exposed to as well as the solvent accessibility of each modification site.

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