Probing Conformational Diversity of Fc Domains in Aggregation-Prone Monoclonal Antibodies
| Autor: | Michael T. Jones, Alphonse Ignatius A, Kimmel M, Subhabrata Majumder |
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| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular 0301 basic medicine Glycosylation Protein Conformation medicine.drug_class Protein domain Pharmaceutical Science Protein aggregation 010402 general chemistry Monoclonal antibody 01 natural sciences Protein Aggregates 03 medical and health sciences chemistry.chemical_compound Protein structure Protein Domains medicine Pharmacology (medical) Nuclear Magnetic Resonance Biomolecular Pharmacology biology Protein Stability Chemistry Organic Chemistry Antibodies Monoclonal Nuclear magnetic resonance spectroscopy Fusion protein Immunoglobulin Fc Fragments 0104 chemical sciences 030104 developmental biology Immunoglobulin G Biophysics biology.protein Molecular Medicine Antibody Biotechnology |
| Zdroj: | Pharmaceutical Research. 35 |
| ISSN: | 1573-904X 0724-8741 |
| DOI: | 10.1007/s11095-018-2500-8 |
| Popis: | Fc domains are an integral component of monoclonal antibodies (mAbs) and Fc-based fusion proteins. Engineering mutations in the Fc domain is a common approach to achieve desired effector function and clinical efficacy of therapeutic mAbs. It remains debatable, however, whether molecular engineering either by changing glycosylation patterns or by amino acid mutation in Fc domain could impact the higher order structure of Fc domain potentially leading to increased aggregation propensities in mAbs. Here, we use NMR fingerprinting analysis of Fc domains, generated from selected Pfizer mAbs with similar glycosylation patterns, to address this question. Specifically, we use high resolution 2D [13C-1H] NMR spectra of Fc fragments, which fingerprints methyl sidechain bearing residues, to probe the correlation of higher order structure with the storage stability of mAbs. Thermal calorimetric studies were also performed to assess the stability of mAb fragments. Unlike NMR fingerprinting, thermal melting temperature as obtained from calorimetric studies for the intact mAbs and fragments (Fc and Fab), did not reveal any correlation with the aggregation propensities of mAbs. Despite >97% sequence homology, NMR data suggests that higher order structure of Fc domains could be dynamic and may result in unique conformation(s) in solution. The overall glycosylation pattern of these mAbs being similar, these conformation(s) could be linked to the inherent plasticity of the Fc domain, and may act as early transients to the overall aggregation of mAbs. |
| Databáze: | OpenAIRE |
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