High-Resolution IMS–MS to Assign Additional Disulfide Bridge Pairing in Complementarity-Determining Regions of an IgG4 Monoclonal Antibody
| Autor: | Elsa Wagner-Rousset, Olivier Colas, Kevin Giles, Sarah Cianférani, Thomas Botzanowski, Evolène Deslignière, Oscar Hernandez-Alba, Alain Beck, Dale Cooper-Shepherd, Hélène Diemer, Guillaume Béchade |
|---|---|
| Přispěvatelé: | Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Infrastructure Nationale de Protéomique, FR2048 ProFI, Waters Corporation, Centre d'Immunologie Pierre Fabre (CIPF), PIERRE FABRE |
| Rok vydání: | 2021 |
| Předmět: |
medicine.drug_class
Complementarity determining region Monoclonal antibody Immunoglobulin light chain 01 natural sciences Mass Spectrometry 03 medical and health sciences chemistry.chemical_compound Structural Biology Ion Mobility Spectrometry medicine Humans [CHIM]Chemical Sciences Disulfides Spectroscopy 030304 developmental biology 0303 health sciences Dipeptide biology Chemistry Immunogenicity 010401 analytical chemistry Antibodies Monoclonal Complementarity Determining Regions 0104 chemical sciences Biochemistry Immunoglobulin G biology.protein Antibody Critical quality attributes Cysteine |
| Zdroj: | Journal of The American Society for Mass Spectrometry Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), In press, ⟨10.1021/jasms.1c00151⟩ Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), In press |
| ISSN: | 1879-1123 1044-0305 |
| DOI: | 10.1021/jasms.1c00151 |
| Popis: | International audience; Monoclonal antibodies (mAbs) have taken on an increasing importance for the treatment of various diseases, including cancers and immunological disorders. Disulfide bonds play a pivotal role in therapeutic antibody structure and activity relationships. Disulfide connectivity and cysteine-related variants are considered as critical quality attributes (CQAs) that must be monitored during mAb manufacturing and storage, as non-native disulfide bridges and aggregates might be responsible for loss of biological function and immunogenicity. The presence of cysteine residues in the Complementarity-Determining Regions (CDRs) is rare in human antibodies but may be critical for the antigen-binding or deleterious for therapeutic antibody development. Consequently, in-depth characterization of their disulfide network is a prerequisite for mAb developability assessment. Mass spectrometry (MS) techniques represent powerful tools for accurate identification of disulfide connectivity. We report here on the MS-based characterization of an IgG4 comprising two additional cysteine residues in the CDR of its light chain. Classical bottom-up approaches after trypsin digestion first allowed identification of a dipeptide containing two disulfide bridges. To further investigate the conformational heterogeneity of the disulfide-bridged dipeptide, we performed ion mobility spectrometry-mass spectrometry (IMS-MS) experiments. Our results highlight benefits of high resolution IMS-MS to tackle the conformational landscape of disulfide peptides generated after trypsin digestion of a humanized IgG4 mAb under development. By comparing arrival time distributions of the mAb collected peptide and synthetic peptides, cyclic IMS afforded unambiguous assessment of disulfide bonds. In addition to classical peptide mapping, qualitative high-resolution IMS-MS can be of great interest to identify disulfide bonds within therapeutic mAbs. |
| Databáze: | OpenAIRE |
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