scholarly journals High-Resolution IMS–MS to Assign Additional Disulfide Bridge Pairing in Complementarity-Determining Regions of an IgG4 Monoclonal Antibody

2021 ◽  
Author(s):  
Evolène Deslignière ◽  
Thomas Botzanowski ◽  
Hélène Diemer ◽  
Dale A. Cooper-Shepherd ◽  
Elsa Wagner-Rousset ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Resolution ◽  
Disulfide Bridge ◽  
Complementarity Determining Regions

scholarly journals A 2.8-Angstrom-Resolution Cryo-Electron Microscopy Structure of Human Parechovirus 3 in Complex with Fab from a Neutralizing Antibody

2018 ◽  
Vol 93 (4) ◽  
Author(s):  
Aušra Domanska ◽  
Justin W. Flatt ◽  
Joonas J. J. Jukonen ◽  
James A. Geraets ◽  
Sarah J. Butcher
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
High Resolution ◽  
Cultured Cells ◽  
Human Monoclonal Antibody ◽  
Neutralizing Antibody ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Human Parechovirus ◽  
Cryo Electron Microscopy

ABSTRACTHuman parechovirus 3 (HPeV3) infection is associated with sepsis characterized by significant immune activation and subsequent tissue damage in neonates. Strategies to limit infection have been unsuccessful due to inadequate molecular diagnostic tools for early detection and the lack of a vaccine or specific antiviral therapy. Toward the latter, we present a 2.8-Å-resolution structure of HPeV3 in complex with fragments from a neutralizing human monoclonal antibody, AT12-015, using cryo-electron microscopy (cryo-EM) and image reconstruction. Modeling revealed that the epitope extends across neighboring asymmetric units with contributions from capsid proteins VP0, VP1, and VP3. Antibody decoration was found to block binding of HPeV3 to cultured cells. Additionally, at high resolution, it was possible to model a stretch of RNA inside the virion and, from this, identify the key features that drive and stabilize protein-RNA association during assembly.IMPORTANCEHuman parechovirus 3 (HPeV3) is receiving increasing attention as a prevalent cause of sepsis-like symptoms in neonates, for which, despite the severity of disease, there are no effective treatments available. Structural and molecular insights into virus neutralization are urgently needed, especially as clinical cases are on the rise. Toward this goal, we present the first structure of HPeV3 in complex with fragments from a neutralizing monoclonal antibody. At high resolution, it was possible to precisely define the epitope that, when targeted, prevents virions from binding to cells. Such an atomic-level description is useful for understanding host-pathogen interactions and viral pathogenesis mechanisms and for finding potential cures for infection and disease.

An improved capillary isoelectric focusing-mass spectrometry method for high-resolution characterization of monoclonal antibody charge variants

2022 ◽  
Author(s):  
Tian Xu ◽  
Linjie Han ◽  
Alayna M. George Thompson ◽  
Liangliang Sun
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Capillary Isoelectric Focusing ◽  
Spectrometry Method ◽  
Mass Spectrometry Method ◽  
Charge Variants

Routine and high-resolution characterization of monoclonal antibody (mAb) charge variants is vital for controlling mAb quality as therapeutics. Capillary isoelectric focusing-mass spectrometry (cIEF-MS) has emerged as a powerful tool for...

Application of high-resolution MS in the quantification of a therapeutic monoclonal antibody in human plasma

Bioanalysis ◽  
2014 ◽  
Vol 6 (13) ◽  
pp. 1767-1779 ◽  
Author(s):  
Kevork Mekhssian ◽  
Jean-Nicholas Mess ◽  
Fabio Garofolo
Keyword(s):  
Monoclonal Antibody ◽  
High Resolution ◽  
Human Plasma ◽  
Therapeutic Monoclonal Antibody

Structure of P46, an immunodominant surface protein from Mycoplasma hyopneumoniae: interaction with a monoclonal antibody

2020 ◽  
Vol 76 (5) ◽  
pp. 418-427
Author(s):  
Alicia Guasch ◽  
Jordi Montané ◽  
Alexandra Moros ◽  
Jaume Piñol ◽  
Marta Sitjà ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Structural Information ◽  
Surface Protein ◽  
Mycoplasma Hyopneumoniae ◽  
Economic Losses ◽  
Fab Fragment ◽  
New Therapeutic Approaches ◽  
Complementarity Determining Regions ◽  
Extracellular Side

Mycoplasma hyopneumoniae is a prokaryotic pathogen that colonizes the respiratory ciliated epithelial cells in swine. Infected animals suffer respiratory lesions, causing major economic losses in the porcine industry. Characterization of the immunodominant membrane-associated proteins from M. hyopneumoniae may be instrumental in the development of new therapeutic approaches. Here, the crystal structure of P46, one of the main surface-antigen proteins, from M. hyopneumoniae is presented and shows N- and C-terminal α/β domains connected by a hinge. The structures solved in this work include a ligand-free open form of P46 (3.1 Å resolution) and two ligand-bound structures of P46 with maltose (2.5 Å resolution) and xylose (3.5 Å resolution) in open and closed conformations, respectively. The ligand-binding site is buried in the cleft between the domains at the hinge region. The two domains of P46 can rotate with respect to each other, giving open or closed alternative conformations. In agreement with this structural information, sequence analyses show similarities to substrate-binding members of the ABC transporter superfamily, with P46 facing the extracellular side as a functional subunit. In the structure with xylose, P46 was also bound to a high-affinity (K d = 29 nM) Fab fragment from a monoclonal antibody, allowing the characterization of a structural epitope in P46 that exclusively involves residues from the C-terminal domain. The Fab structure in the complex with P46 shows only small conformational rearrangements in the six complementarity-determining regions (CDRs) with respect to the unbound Fab (the structure of which is also determined in this work at 1.95 Å resolution). The structural information that is now available should contribute to a better understanding of sugar nutrient intake by M. hyopneumoniae. This information will also allow the design of protocols and strategies for the generation of new vaccines against this important swine pathogen.

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