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Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
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Gas-phase unfolding assay rapidly predicts structure-function relationships in engineered antibodies with tuned

Rosendo C Villafuerte-Vega1,2, Michael R Armbruster1, Hayden Fisher3,4

  • 1Department of Chemistry, University of Michigan, Ann Arbor, MI, USA.

Nature Communications
|June 29, 2026
PubMed

Human (h)IgG2 monoclonal antibodies (mAbs) are potent agonists due, in part, to their ability to undergo disulfide shuffling within their hinge regions. Herein, we describe a rapid, sensitive, collision-induced unfolding (CIU) assay that possesses a predictive relationship between gas-phase protein unfolding and agonism in hIgG2 variants. Furthermore, our results highlight the significance of hinge engineering in tuning mAb structure-function relationships for the development of future biotherapeutics.

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