Backbone 1H, 13C, and 15N resonance assignments of the Fc fragment of human immunoglobulin G glycoprotein

The Fc portion of immunoglobulin G (IgG) recruits complements and its cognate receptors, thereby promoting defensive mechanisms in the humoral immune system. These effector functions critically depend on N-glycosylation at the Fc region, which is therefore regarded as a crucial factor in the design and production of therapeutic antibodies. NMR spectroscopy plays a unique role in the characterization of conformational dynamics and intermolecular interactions of IgG-Fc in solutions. Here, we report NMR assignments of the glycosylated Fc fragment (Mr 53 kDa), cleaved from a chimeric antibody with human IgG1 constant regions, which was produced in Chinese hamster ovary cells with uniform 13C- and 15N-labeling.

• Publication year

  2014

• Venue

  Biomolecular NMR Assignments

• Publication date

  2014-10-08

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1007/s12104-014-9586-7PMID 25291979PMCID 4568019
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Analytical tools for characterizing biopharmaceuticals and the implications for biosimilars

  2012cited by this paper
• Unique carbohydrate–carbohydrate interactions are required for high affinity binding between FcγRIII and antibodies lacking core fucose

  2011cited by this paper
• Elucidation of Acid-induced Unfolding and Aggregation of Human Immunoglobulin IgG1 and IgG2 Fc

  2011cited by this paper
• Advances in the assessment and control of the effector functions of therapeutic antibodies

  2011cited by this paper
• Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans

  2011cited by this paper
• Stable-isotope-assisted NMR approaches to glycoproteins using immunoglobulin G as a model system.

  2010cited by this paper
• Dynamics and interactions of glycoconjugates probed by stable-isotope-assisted NMR spectroscopy.

  2010influential reference
• Structural and molecular basis for hyperspecificity of RNA aptamer to human immunoglobulin G.

  2008cited by this paper
• Assignment of backbone 1H, 13C and 15N resonances of human IgG1 Fc (51.4 kDa)

  2007cited by this paper
• 3.32 – Antibody Structures

  2007cited by this paper
• Structural comparison of fucosylated and nonfucosylated Fc fragments of human immunoglobulin G1.

  2007cited by this paper
• Glycoform-dependent conformational alteration of the Fc region of human immunoglobulin G1 as revealed by NMR spectroscopy.

  2006influential reference
• The CCPN data model for NMR spectroscopy: Development of a software pipeline

  2005cited by this paper
• Structural analysis of human IgG-Fc glycoforms reveals a correlation between glycosylation and structural integrity.

  2003cited by this paper
• C NMR study of the mode of interaction in solution of the B fragment of staphylococcal protein A and the Fc fragments of mouse immunoglobulin

  2001cited by this paper
• Model for the complex between protein G and an antibody Fc fragment in solution.

  1995cited by this paper
• Proteolytic fragmentation with high specificity of mouse immunoglobulin G. Mapping of proteolytic cleavage sites in the hinge region.

  1995cited by this paper
• NMRPipe: A multidimensional spectral processing system based on UNIX pipes

  1995cited by this paper
• Dynamical structure of the hinge region of immunoglobulin G as studied by 13C nuclear magnetic resonance spectroscopy.

  1994cited by this paper
• 13C NMR study of the mode of interaction in solution of the B fragment of staphylococcal protein A and the Fc fragments of mouse immunoglobulin G

  1993cited by this paper
• Complete assignment of the methionyl carbonyl carbon resonances in switch variant anti-dansyl antibodies labeled with [1-13C]methionine.

  1991cited by this paper
• Carbon-13 NMR study of switch variant anti-dansyl antibodies: antigen binding and domain-domain interactions.

  1991cited by this paper

• Unlabeled NMR Approach with Site-Specific Methyl Assignments for Structural Evaluation of the IgG1 Fc Region.

  2026cites this paper
• Exploring glycoform-dependent dynamic modulations in human immunoglobulin G via computational and experimental approaches

  2025cites this paper
• Assessment of ADC Higher Order Structure Through 2D NMR Analysis

  2025influential citation
• Toward Site-Specific Characterization of Structural Perturbations on Glycosylated Fc Using NMR at Natural Abundance

  2025cites this paper
• A fast and efficient strategy for the NMR assignment of Fab methyl groups

  2025cites this paper
• NMR investigations of glycan conformation, dynamics, and interactions.

  2024cites this paper
• One N-glycan regulates natural killer cell antibody-dependent cell-mediated cytotoxicity and modulates Fc γ receptor IIIa/CD16a structure

  2024cites this paper
• One N-glycan regulates natural killer cell antibody-dependent cell-mediated cytotoxicity and modulates Fc γ receptor IIIa / CD16a structure

  2024cites this paper
• Enabling site-specific NMR investigations of therapeutic Fab using a cell-free based isotopic labeling approach: application to anti-LAMP1 Fab

  2024cites this paper
• Negative interference with antibody-dependent cellular cytotoxicity mediated by rituximab from its interactions with human serum proteins

  2023cites this paper
• Backbone NMR assignment of the yeast expressed Fab fragment of the NISTmAb reference antibody

  2023cites this paper
• Comprehensive characterization of higher order structure changes in methionine oxidized monoclonal antibodies via NMR chemometric analysis and biophysical approaches

  2023cites this paper
• Integration of NMR Spectroscopy in an Analytical Workflow to Evaluate the Effects of Oxidative Stress on Abituzumab: Beyond the Fingerprint of mAbs

  2023cites this paper
• Expression of 2H, 13C, 15N-labeled NIST-Fab fragment in the methylotrophic yeast Komagataella phaffii for nuclear magnetic resonance studies

  2023cites this paper
• Metabolic15N labeling of the N-glycosylated immunoglobulin G1 Fc with an engineered Saccharomyces cerevisiae strain

  2022cites this paper
• Specific location of galactosylation in an afucosylated antiviral monoclonal antibody affects its FcγRIIIA binding affinity

  2022cites this paper
• Glutamine-free mammalian expression of recombinant glycoproteins with uniform isotope labeling: an application for NMR analysis of pharmaceutically relevant Fc glycoforms of human immunoglobulin G1

  2022cites this paper
• Structural Fingerprints of an Intact Monoclonal Antibody Acquired under Formulated Storage Conditions via 15N Direct Detection Nuclear Magnetic Resonance.

  2020cites this paper
• Silkworm Pupae Function as Efficient Producers of Recombinant Glycoproteins with Stable-Isotope Labeling

  2020cites this paper
• Role of NMR in High Ordered Structure Characterization of Monoclonal Antibodies

  2020cites this paper
• Utility of High Resolution NMR Methods to Probe the Impact of Chemical Modifications on Higher Order Structure of Monoclonal Antibodies in Relation to Antigen Binding

  2019cites this paper
• Assessment of the higher order structure of Humira®, Remicade®, Avastin®, Rituxan®, Herceptin®, and Enbrel® by 2D‐NMR fingerprinting

  2019cites this paper
• The Preparation and Solution NMR Spectroscopy of Human Glycoproteins Is Accessible and Rewarding.

  2019cites this paper
• A synopsis of recent developments defining how N-glycosylation impacts immunoglobulin G structure and function.

  2019cites this paper
• Stable isotope labeling approaches for NMR characterization of glycoproteins using eukaryotic expression systems

  2018cites this paper
• Stable isotope labeling approaches for NMR characterization of glycoproteins using eukaryotic expression systems

  2018cites this paper
• Technical Basis for Nuclear Magnetic Resonance Approach for Glycoproteins

  2018cites this paper
• [Structural Biological Approach to Biopharmaceuticals].

  2018cites this paper
• Structure and Dynamics of Immunoglobulin G Glycoproteins.

  2018cites this paper
• Probing Conformational Diversity of Fc Domains in Aggregation-Prone Monoclonal Antibodies

  2018cites this paper
• The recognition of glycans by protein receptors. Insights from NMR spectroscopy.

  2018cites this paper
• SAR News No.35

  2018cites this paper
• NMR Detection of Semi-Specific Antibody Interactions in Serum Environments

  2017cites this paper
• Elucidation of potential sites for antibody engineering by fluctuation editing

  2017cites this paper
• Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy.

  2017cites this paper
• NMR Characterization of the Dynamic Conformations of Oligosaccharides

  2017cites this paper
• Biosimilar structural comparability assessment by NMR: from small proteins to monoclonal antibodies

  2016cites this paper
• Structural basis of the molecular mechanisms underlying intracellular quality control of glycoproteins mediated by their glucosylation

  2016cites this paper
• The Structural Role of Ant ibody N-Glycosylation in Receptor Interactions Graphical Abstract Highlights

  2015cites this paper
• Stable isotope labeling of glycoprotein expressed in silkworms using immunoglobulin G as a test molecule

  2015cites this paper
• The Structural Role of Antibody N-Glycosylation in Receptor Interactions.

  2015cites this paper
• A Hybrid Strategy for the Preparation of 13C-labeled High-mannose-type Oligosaccharides with Terminal Glucosylation for NMR Study

  2015cites this paper
• High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension.

  2015cites this paper
• NMR-based structural validation of therapeutic antibody produced in Nicotiana benthamiana

  2015cites this paper