Gas-phase unfolding assay rapidly predicts structure-function relationships in engineered antibodies with tuned flexibilities.

Villafuerte-Vega, Rosendo C; Armbruster, Michael R; Fisher, Hayden; Elliott, Isabel G; Chan, H T Claude; Inzhelevskaya, Tatyana; Mockridge, C Ian; Penfold, Christine A et al. · Nat Commun · 2026

basic_science · Level V

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Abstract

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.