Articles in the Category ”References”
FabULOUS™ in HIV Vaccine Research
The Fc N-glycosylation profile can be linked to non-neutralizing functions in a vaccine. In a recent study by Chen et al., FabULOUS digestion provided valuable insights in HIV vaccine research following N-glycosylation analysis of the Fc region in mouse antibodies.
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Localization of O-glycan Sites in CD24Fc by OpeRATOR® Digestion and LC-MS Analysis
Scientists from Merck & Co. and Glycotope describe the localization of 12 O-glycosylation sites in the heavily O-glycosylated fusion protein CD24Fc by digestion with the O-glycoprotease OpeRATOR in combination with LC-MS analysis. Two different workflows were developed, one based on fractionation in combination with LC-MS/MS CID and one based on LC-MS/MS EThcD. OpeRATOR digestion enabled the localization of O-glycan sites in both workflows and relative quantification of different glycoforms.
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FabRICATOR® in Correlative N-glycan Analysis of Cetuximab Produced in Different Expression Systems
A strategy for comparative characterization of cetuximab produced in a variety of expression cell lines is described in this paper from scientists at the National Institute for Bioprocessing Research and Training (NIBRT) in Dublin, Ireland. FabRICATOR (IdeS) is a critical tool in this study, allowing for site-specific glycan analysis to be performed following antibody digestion into F(ab’)2 and Fc/2 subunits.
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Unravelling Structural Conformers within a Therapeutic Multispecific mAb using FabALACTICA
Scientists at the University of Strasbourg and Sanofi describe the global structural characterisation of a trispecific antibody which exhibits two distinct structural isoforms when analyzed by size-exclusion chromatography (SEC). FabALACTICA proved to be a valuable tool in this study as above hinge digestion of this trispecific antibody allowed for middle-level characterisation of the isomeric Fab regions and facilitated cyclic-ion mobility (cIM) differentiation of the isomers, which was not possible in the intact molecule without prior SEC separation.
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FabRICATOR® in Middle-Up Quantitative N-Glycan Profiling of Therapeutic Monoclonal Antibodies
A middle-level approach to glycan characterisation is described in this paper by scientists at the University of Geneva and collaborators. FabRICATOR (IdeS), a critical tool in this study, is used to generate Fd’, LC and the glycan-containing Fc subunits, which can be chromatographically separated using HILIC and characterised using mass spectrometry.
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SialEXO® 23 in a Study of Osteoclast Fusion
In a recent paper from Johns Hopkins University School of Medicine in Baltimore, removal of sialic acids from toll-like receptor 2 using SialEXO 23 was found to inhibit osteoclast fusion. These new findings in osteoclast recognition signaling are important for understanding the pathogenesis of different skeletal disorders, also showcasing the versatility of Genovis SmartEnzymes!
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FabRICATOR® in an Inline Electrochemical Reduction Workflow
An inline electrochemical reduction workflow for antibodies coupled with LC-MS analysis is described in this paper from the National Institute for Bioprocessing Research and Training in Dublin. The FabRICATOR enzyme (IdeS) is a key component in the subunit analysis by digesting antibodies at a single site below the hinge, generating a homogenous pool of F(ab’)2 and Fc subunits.
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SialEXO® in a Glycoform Analysis of Intact Erythropoietin
In this study, scientists at Leiden University Medical Center present a MALDI FT-ICR MS method for the glycosylation profiling of intact erythropoietin (EPO). The sialidase mix SialEXO significantly reduced the mass spectrum complexity and facilitated the interpretation of the complex spectra of intact EPO.
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GlycINATOR® in Peptide Mapping of Fc Domain-Drug Conjugates
The field of antibody-drug conjugates (ADCs) is rapidly emerging as a new, important class of cancer therapeutics. ADCs consist of monoclonal antibodies (mAbs) covalently linked to cytotoxic drugs, and after selective surface receptor binding on cancer cells, the drug is delivered into the intracellular compartment and can exert its cytotoxic effect.
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FabALACTICA® in a Quantitative IgG1 Clone Profiling
Our immune system protects us from pathogens and various diseases, and the immune response is dependent on a person’s repertoire of immune cells, antibodies and circulating plasma proteins. A detailed characterization of these plasma components is important for the understanding of how they affect each individual’s immune response.
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