Article
Biochemistry & Molecular Biology
Kyle W. Anderson, Christina Bergonzo, Kerry Scott, Ioannis L. Karageorgos, Elyssia S. Gallagher, Venkata S. Tayi, Michael Butler, Jeffrey W. Hudgens
Summary: Previous studies have proposed different models for stabilizing the Fc-Fc gamma RI immune complex. This study used multiple experimental methods to investigate the role of glycoprotein interactions between IgG1 glycans and the FG-loop of Fc gamma RIa in complex stabilization. The results showed that both intermolecular and intramolecular glycoprotein bonds contribute to the stability of the complex.
JOURNAL OF MOLECULAR BIOLOGY
(2022)
Article
Chemistry, Analytical
Alexander B. Schwahn, Julia Baek, Shanhua Lin, Christopher A. Pohl, Ken Cook
Summary: Characterization and quality control of biotherapeutic proteins often require multiple orthogonal separation techniques, leading to tedious method development. This study demonstrates the use of a linear controlled pH gradient system and universal buffer solutions to improve efficiency and minimize manual intervention in method development and analysis.
ANALYTICAL CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Christina Bergonzo, J. Todd Hoopes, Zvi Kelman, D. Travis Gallagher
Summary: In this study, all-atom simulations were used to compare the dynamic behavior of four different structural variants of Fc antibody domain. The effects of hinge and glycans on stability were independently measured using umbrella sampling simulations. Principal component analysis was conducted to determine the free energy effects along the dominant mode of motion for Fc.
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS
(2023)
Article
Plant Sciences
Gernot Beihammer, Julia Koenig-Beihammer, Benjamin Kogelmann, Valentina Ruocco, Clemens Gruenwald-Gruber, Marc-Andre D'Aoust, Pierre-Olivier Lavoie, Pooja Saxena, Johannes S. S. Gach, Herta Steinkellner, Richard Strasser
Summary: By testing a single-subunit oligosaccharyltransferase from Leishmania donovani (LdOST), we found that it can significantly increase the Fc glycosylation level of IgG1 without changing the overall N-glycan composition. This finding is important for reducing the heterogeneity of plant-produced antibodies and improving their stability and effector functions.
FRONTIERS IN PLANT SCIENCE
(2023)
Article
Immunology
Somanath Kallolimath, Roman Palt, Esther Foederl-Hoebenreich, Lin Sun, Qiang Chen, Florian Pruckner, Lukas Eidenberger, Richard Strasser, Kurt Zatloukal, Herta Steinkellner
Summary: This study successfully produced pentameric IgM antibodies in glycoengineered plants that carry a highly reproducible human-type N-glycosylation profile. The pentameric IgMs showed increased antigen binding and virus neutralization potency, up to 390-fold, compared to the parental IgG1.
FRONTIERS IN IMMUNOLOGY
(2023)
Article
Biochemical Research Methods
Jun-Jie Yuan, Dong Gao, Feng Hu, Yang Shi, Zhen-Hua Wu, Chuan-Qin Hu, Xiao-Dong Huang, Wei-Jie Fang, Hai-Tao Zhang, Hai-Bin Wang
Summary: This research investigated and characterized charge variants in monoclonal antibodies, identifying the main causes and effects. Through extensive analysis, it was concluded that charge heterogeneity does not impact biological functions. The study suggests that charge variants should be considered effective components, unless there is a significant impact on biological functions.
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
(2021)
Review
Chemistry, Analytical
Harleen Kaur, Jeff Beckman, Yiting Zhang, Zheng Jian Li, Marton Szigeti, Andras Guttman
Summary: Capillary electrophoresis is a powerful technique for comprehensive physicochemical characterization of therapeutic biomolecules, such as monoclonal antibodies, providing high resolution separation and high sensitivity characterization. Techniques like SDS-CGE, CZE, and iCIEF are increasingly used to assess heterogeneity in mAbs and related therapeutic modalities, with advantages and disadvantages that are critically discussed.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Albert Bondt, Max Hoek, Sem Tamara, Bastiaan de Graaf, Weiwei Peng, Douwe Schulte, Danique M. H. van Rijswijck, Maurits A. den Boer, Jean-Francois Greisch, Meri R. J. Varkila, Joost Snijder, Olaf L. Cremer, Marc J. M. Bonten, Albert J. R. Heck
Summary: The study revealed that the circulating IgG1 repertoire in human plasma is dominated by a limited number of clones in healthy donors and septic patients. Each individual donor exhibits a unique serological IgG1 repertoire, which remains stable over time but can adapt rapidly to changes in physiology. Conducting de novo sequencing at the protein level is important for understanding the high mutation rates observed in IgG1 clones.
Article
Pharmacology & Pharmacy
Alexie Mayor, Beatrice Thibert, Sylvain Huille, Fethi Bensaid, Renaud Respaud, Heloise Audat, Nathalie Heuze-Vourc'h
Summary: In the past decade, oral inhalation has been a popular research topic for direct treatment of respiratory diseases. The formulation of inhaled antibodies plays a crucial role in their stability and lung safety. This study reveals the significant impact of the buffering system on the stability of monoclonal antibodies during nebulization.
EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS
(2022)
Article
Biology
Simona Saporiti, Tommaso Laurenzi, Uliano Guerrini, Crescenzo Coppa, Wolf Palinsky, Giulia Benigno, Luca Palazzolo, Omar Ben Mariem, Linda Montavoci, Mara Rossi, Fabio Centola, Ivano Eberini
Summary: N-glycosylation and light chain isotype play important roles in modulating the conformational behavior of monoclonal antibodies. Through accelerated molecular dynamics simulations, this study reveals how fucosylation and light chain isotype influence the hinge behavior, Fc conformation, and glycan chain position, factors that potentially affect the binding to Fc gamma receptors. Moreover, this work demonstrates the suitability of accelerated molecular dynamics as an approach to elucidate experimental results on antibody conformation.
COMMUNICATIONS BIOLOGY
(2023)
Article
Pharmacology & Pharmacy
Yue Liu, Hao Li, Zhen Yan, Lianshan Zhang, Piaoyang Sun
Summary: This study explores the mechanism of CDR fragmentation in therapeutic antibodies and identifies excipients that can control fragmentation. Trp100 oxidation is found to be correlated with fragment formation, and adding Trp can reduce the level of fragmentation. Moreover, substituting polysorbate 80 (PS80) with other surfactants, using a combination of antioxidants, or combining EDTA with PS80 can maintain the stability of the antibody.
EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS
(2022)
Article
Multidisciplinary Sciences
Sofia B. Carvalho, Ricardo A. Gomes, Anja Pfenninger, Martina Fischer, Michaela Strotbek, Ines A. Isidro, Nihal Tugcu, Patricia Gomes-Alves
Summary: As the requirements of regulatory agencies for the quality of biopharmaceuticals increase, the use of mass spectrometry-based methods such as the multi-attribute method (MAM) has become a valuable tool for quality control. MAM allows for the direct and simultaneous monitoring of critical product quality attributes using high-quality mass spectrometry data, replacing traditional orthogonal methods and reducing time and costs. This article describes the implementation of MAM using a QTOF high-resolution platform and discusses sample preparation and LC-MS parameters troubleshooting for batch analysis and high-throughput applications.
Article
Biotechnology & Applied Microbiology
Yuanli Song, Hui Cai, Zhijun Tan, Nesredin Mussa, Zheng-Jian Li
Summary: This study demonstrates that therapeutic IgG1 and IgG4 antibodies display different sensitivity to the reducing agent TCEP and follow different pathways to form low molecular weight (LMW) species. The findings provide mechanistic insights into LMW formation of IgG1 and IgG4, which have implications for the selection of IgG1 or IgG4 in bispecific antibodies and cysteine-based antibody-drug conjugates.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Medicine, Research & Experimental
Yilue Zhang, Christian Schoeneich
Summary: In this study, a novel visible light-induced heavy chain fragmentation method of IgG1 mediated by an Fe(III)-containing histidine buffer was reported. The fragmentation involves oxidative cleavage and intrachain radical reactions. The presence of His is required for the cleavage, and it can be prevented by EDTA.
MOLECULAR PHARMACEUTICS
(2022)
Article
Medicine, Research & Experimental
Zhou Tian, Feng Qian
Summary: The nature of ATP as amphiphilic and hydrotrope plays a role in maintaining solubility and preventing liquid-liquid phase separation of proteins, with ATP-MA1 static interaction driving phase separation. The attractive protein-protein interaction exclusively increased in the presence of ATP, altering protein surface properties and inducing LLPS of MA1. This ATP-induced LLPS was effectively eliminated by Mg2+ chelation with ATP to negate the static interaction.
MOLECULAR PHARMACEUTICS
(2021)