Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy

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Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy. / Jensen, Rasmus K; Jabs, Frederic; Miehe, Michaela; Mølgaard, Brian; Pfützner, Wolfgang; Möbs, Christian; Spillner, Edzard; Andersen, Gregers R.

In: Allergy: European Journal of Allergy and Clinical Immunology, Vol. 75, No. 8, 08.2020.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Jensen, RK, Jabs, F, Miehe, M, Mølgaard, B, Pfützner, W, Möbs, C, Spillner, E & Andersen, GR 2020, 'Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy', Allergy: European Journal of Allergy and Clinical Immunology, vol. 75, no. 8. https://doi.org/10.1111/all.14222

APA

Jensen, R. K., Jabs, F., Miehe, M., Mølgaard, B., Pfützner, W., Möbs, C., Spillner, E., & Andersen, G. R. (2020). Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy. Allergy: European Journal of Allergy and Clinical Immunology, 75(8). https://doi.org/10.1111/all.14222

CBE

Jensen RK, Jabs F, Miehe M, Mølgaard B, Pfützner W, Möbs C, Spillner E, Andersen GR. 2020. Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy. Allergy: European Journal of Allergy and Clinical Immunology. 75(8). https://doi.org/10.1111/all.14222

MLA

Vancouver

Jensen RK, Jabs F, Miehe M, Mølgaard B, Pfützner W, Möbs C et al. Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy. Allergy: European Journal of Allergy and Clinical Immunology. 2020 Aug;75(8). https://doi.org/10.1111/all.14222

Author

Jensen, Rasmus K ; Jabs, Frederic ; Miehe, Michaela ; Mølgaard, Brian ; Pfützner, Wolfgang ; Möbs, Christian ; Spillner, Edzard ; Andersen, Gregers R. / Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy. In: Allergy: European Journal of Allergy and Clinical Immunology. 2020 ; Vol. 75, No. 8.

Bibtex

@article{c6193f014858429eac94951fd2dbd86c,
title = "Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy",
abstract = "Background: IgE is the central antibody isotype in TH2-biased immunity and allergic diseases. The structure of intact IgE and the impact of IgE-targeting molecules on IgE however remain elusive. In order to obtain insights into IgE biology and the clinical impact, we aimed for structure determination of IgE and the complex of IgE with the anti-IgE antibody ligelizumab. Methods: Structures of two distinct intact IgE with specificity for cross-reactive carbohydrate determinants and Der p 2 as well as complexes of ligelizumab-Fab with IgE and IgE Fc were assessed by negative stain electron microscopy and solution scattering. Inhibition of IgE binding and displacement of receptor-bound IgE were assessed using cellular assays, basophil activation testing and ELIFAB assays. Results: Our data reveal that the investigated IgE molecules share an overall rigid conformation. In contrast to the IgE Fc fragment, the IgE Fc in intact IgE is significantly less asymmetrically bent. The proximal and the distal Fabs are rigidly tethered to the Fc. Binding of ligelizumab to IgE in a 2:1 stoichiometry induces an extended and twofold symmetrical conformation of IgE, which retains a rigid Fab-Fc architecture. Analyses of effector cell activation revealed that ligelizumab inhibits IgE binding without displacing receptor-bound IgE. Together with an interference of CD23 binding, the data underline a functional activity similar to omalizumab. Conclusions: Our data reveal the first structures of intact IgE suggesting that the IgE Fab is fixed relative to the Fc. Furthermore, we provide a structural rationale for the inhibitory mechanism of ligelizumab.",
keywords = "IgE, antibody structure, electron microscopy, flexibility, therapeutic antibody",
author = "Jensen, {Rasmus K} and Frederic Jabs and Michaela Miehe and Brian M{\o}lgaard and Wolfgang Pf{\"u}tzner and Christian M{\"o}bs and Edzard Spillner and Andersen, {Gregers R}",
note = "This article is protected by copyright. All rights reserved.",
year = "2020",
month = aug,
doi = "10.1111/all.14222",
language = "English",
volume = "75",
journal = "Allergy: European Journal of Allergy and Clinical Immunology",
issn = "0105-4538",
publisher = "Wiley Online",
number = "8",

}

RIS

TY - JOUR

T1 - Structure of intact IgE and the mechanism of ligelizumab revealed by electron microscopy

AU - Jensen, Rasmus K

AU - Jabs, Frederic

AU - Miehe, Michaela

AU - Mølgaard, Brian

AU - Pfützner, Wolfgang

AU - Möbs, Christian

AU - Spillner, Edzard

AU - Andersen, Gregers R

N1 - This article is protected by copyright. All rights reserved.

PY - 2020/8

Y1 - 2020/8

N2 - Background: IgE is the central antibody isotype in TH2-biased immunity and allergic diseases. The structure of intact IgE and the impact of IgE-targeting molecules on IgE however remain elusive. In order to obtain insights into IgE biology and the clinical impact, we aimed for structure determination of IgE and the complex of IgE with the anti-IgE antibody ligelizumab. Methods: Structures of two distinct intact IgE with specificity for cross-reactive carbohydrate determinants and Der p 2 as well as complexes of ligelizumab-Fab with IgE and IgE Fc were assessed by negative stain electron microscopy and solution scattering. Inhibition of IgE binding and displacement of receptor-bound IgE were assessed using cellular assays, basophil activation testing and ELIFAB assays. Results: Our data reveal that the investigated IgE molecules share an overall rigid conformation. In contrast to the IgE Fc fragment, the IgE Fc in intact IgE is significantly less asymmetrically bent. The proximal and the distal Fabs are rigidly tethered to the Fc. Binding of ligelizumab to IgE in a 2:1 stoichiometry induces an extended and twofold symmetrical conformation of IgE, which retains a rigid Fab-Fc architecture. Analyses of effector cell activation revealed that ligelizumab inhibits IgE binding without displacing receptor-bound IgE. Together with an interference of CD23 binding, the data underline a functional activity similar to omalizumab. Conclusions: Our data reveal the first structures of intact IgE suggesting that the IgE Fab is fixed relative to the Fc. Furthermore, we provide a structural rationale for the inhibitory mechanism of ligelizumab.

AB - Background: IgE is the central antibody isotype in TH2-biased immunity and allergic diseases. The structure of intact IgE and the impact of IgE-targeting molecules on IgE however remain elusive. In order to obtain insights into IgE biology and the clinical impact, we aimed for structure determination of IgE and the complex of IgE with the anti-IgE antibody ligelizumab. Methods: Structures of two distinct intact IgE with specificity for cross-reactive carbohydrate determinants and Der p 2 as well as complexes of ligelizumab-Fab with IgE and IgE Fc were assessed by negative stain electron microscopy and solution scattering. Inhibition of IgE binding and displacement of receptor-bound IgE were assessed using cellular assays, basophil activation testing and ELIFAB assays. Results: Our data reveal that the investigated IgE molecules share an overall rigid conformation. In contrast to the IgE Fc fragment, the IgE Fc in intact IgE is significantly less asymmetrically bent. The proximal and the distal Fabs are rigidly tethered to the Fc. Binding of ligelizumab to IgE in a 2:1 stoichiometry induces an extended and twofold symmetrical conformation of IgE, which retains a rigid Fab-Fc architecture. Analyses of effector cell activation revealed that ligelizumab inhibits IgE binding without displacing receptor-bound IgE. Together with an interference of CD23 binding, the data underline a functional activity similar to omalizumab. Conclusions: Our data reveal the first structures of intact IgE suggesting that the IgE Fab is fixed relative to the Fc. Furthermore, we provide a structural rationale for the inhibitory mechanism of ligelizumab.

KW - IgE

KW - antibody structure

KW - electron microscopy

KW - flexibility

KW - therapeutic antibody

U2 - 10.1111/all.14222

DO - 10.1111/all.14222

M3 - Journal article

C2 - 32037590

VL - 75

JO - Allergy: European Journal of Allergy and Clinical Immunology

JF - Allergy: European Journal of Allergy and Clinical Immunology

SN - 0105-4538

IS - 8

ER -