"Small is beautiful"-Examining reliable determination of low-abundant therapeutic antibody glycovariantsOA
"Small is beautiful"-Examining reliable determination of low-abundant therapeutic antibody glycovariants
Glycans associated with biopharmaceutical drugs play crucial roles in drug safety and efficacy,and therefore,their reliable detection and quantification is essential.Our study introduces a multi-level quantification approach for glycosylation analysis in monoclonal antibodies(mAbs),focusing on minor abundant glycovariants.Mass spectrometric data is evaluated mainly employing open-source software tools.Released N-glycan and glycopeptide data form the basis for integrating information across different structural levels up to intact glycoproteins.Comprehensive comparison showed that indeed,variations across structural levels were observed especially for minor abundant species.Utilizing modification finder(MoFi),a tool for annotating mass spectra of intact proteins,we quantify isobaric glycosylation variants at the intact protein level.Our workflow's utility is demonstrated on NISTmAb,rituximab and adalimumab,profiling their minor abundant variants for the first time across diverse structural levels.This study enhances understanding and accessibility in glycosylation analysis,spotlighting minor abundant glycovariants in therapeutic antibodies.
Katharina Böttinger;Christof Regl;Veronika Schäpertöns;Erdmann Rapp;Therese Wohlschlager;Christian G.Huber
Department of Biosciences and Medical Biology,Bioanalytical Research Labs,University of Salzburg,Salzburg,5020,AustriaDepartment of Biosciences and Medical Biology,Bioanalytical Research Labs,University of Salzburg,Salzburg,5020,Austria||Center for Tumorbiology and Immunology(CTBI),University of Salzburg,Salzburg 5020,AustriaglyXera GmbH,Magdeburg,Sachsen-Anhalt,39014,Germany||Max Planck Institute for Dynamics of Complex Technical Systems,Magdeburg,39106,Germany
GlycosylationMass spectrometryMonoclonal antibodiesAbundance profilingMinor glycovariantsMulti-level analysisData integration
《药物分析学报(英文)》 2024 (010)
1435-1449 / 15
Acknowledgments go to Simon Obermaier for initial data treatment as part of his Bachelor thesis.The authors would furthermore like to acknowledge Wolfgang Esser-Skala for providing fragquaxi,MoFi,and CAFOG to the scientific community.This research was funded in whole or in part by the Austrian Sci-ence Fund(FWF)(FG 1201 N,"DigiTherapeutX"],.Figures were created in Inkscape(v1.3.1).Fig.S2 provides an explanation of the used symbol nomenclature and monosaccharide abbreviations.
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