Gas-phase unfolding assay rapidly predicts structure-function relationships in engineered antibodies with tuned flexibilities.
Villafuerte-Vega RC., Armbruster MR., Fisher H., Elliott IG., Chan HTC., Inzhelevskaya T., Mockridge CI., Penfold CA., Duriez PJ., Cragg MS., Ruotolo BT.
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.