Directed evolution of and structural insights into antibody-mediated disruption of a stable receptor-ligand complex

• Published in: Nature communications Vol. 12; no. 1; p. 7069
• Main Authors: Pennington, Luke F., Gasser, Pascal, Kleinboelting, Silke, Zhang, Chensong, Skiniotis, Georgios, Eggel, Alexander, Jardetzky, Theodore S.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 03.12.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 101/28; 13/1; 13/31; 13/95; 631/250/2520; 631/61/338/469; 82/103; 82/80; 82/83; Affinity; Allosteric properties; Animals; Antibodies; BASIC BIOLOGICAL SCIENCES; Cell Membrane; Clinical trials; Coordination compounds; Cryoelectron Microscopy; Crystallography, X-Ray; Directed evolution; Disruption; Effector cells; Engineers; Fc receptors; Healthy Volunteers; Humanities and Social Sciences; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin E - metabolism; Immunoglobulin E - ultrastructure; Immunosuppressive agents; Ligands; Macromolecules; Mice; Mice, Transgenic; Monoclonal antibodies; multidisciplinary; Omalizumab - genetics; Omalizumab - pharmacology; Omalizumab - therapeutic use; Primary Cell Culture; Protein Engineering; Receptors; Receptors, IgE - metabolism; Receptors, IgE - ultrastructure; Science; Science (multidisciplinary); Sf9 Cells; Spodoptera; Yeast; Yeasts
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-27397-z

Antibody drugs exert therapeutic effects via a range of mechanisms, including competitive inhibition, allosteric modulation, and immune effector mechanisms. Facilitated dissociation is an additional mechanism where antibody-mediated “disruption” of stable high-affinity macromolecular complexes can potentially enhance therapeutic efficacy. However, this mechanism is not well understood or utilized therapeutically. Here, we investigate and engineer the weak disruptive activity of an existing therapeutic antibody, omalizumab, which targets IgE antibodies to block the allergic response. We develop a yeast display approach to select for and engineer antibody disruptive efficiency and generate potent omalizumab variants that dissociate receptor-bound IgE. We determine a low resolution cryo-EM structure of a transient disruption intermediate containing the IgE-Fc, its partially dissociated receptor and an antibody inhibitor. Our results provide a conceptual framework for engineering disruptive inhibitors for other targets, insights into the failure in clinical trials of the previous high affinity omalizumab HAE variant and anti-IgE antibodies that safely and rapidly disarm allergic effector cells. Facilitated dissociation is a mechanism where antibody-mediated disruption of high-affinity complexes can enhance the therapeutic effects of a drug. Here the authors present a yeast display approach to select and engineer omalizumab variants that dissociate receptor-bound IgE to accelerate its inhibition of the allergic response.