Articles in the Category ”Applications”

TransGLYCIT™ in a Study on Antibody Characterization

Scientists from the Wistar Institute in Philadelphia and collaborators have studied the IgG N-glycan structures and inflammatory aging markers of people living with and without HIV to determine a potential structure-function relationship. In this study, TransGLYCIT was used to generate anti-HIV antibodies with defined N-glycan structures to study Fc-mediated immune activities in vitro.
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GlyCLICK® Enhances Development of Immuno-SPECT/Fluorescent Tracer for PD-L1 in Preclinical Models

Scientists at Université de Bourgogne and collaborators have described the development of a bimodal tracer targeting PD-L1 in human and murine models for preclinical studies. Using a [111In]-DOTA-aza-BODIPY bimodal probe following conjugation onto PD-L1 monoclonal antibodies (mAbs) through either site-specific or random methods, the ability of the bimodal antibodies to selectively identify PD-L1+ tumors were assessed using both fluorescence and SPECT imaging. There were significant benefits observed with the probe generated using the GlyCLICK site-specific conjugation method compared to a random lysine conjugation approach!
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Mechanistic Insights to Drive Rational Design of Masked Antibodies using FabALACTICA®

Scientists at the University of Cambridge, Astra Zeneca and collaborators undertook mechanistic studies to investigate antibody masking and better understand the parameters involved and their relative impact. FabALACTICA proved to be a valuable tool in this study, allowing the authors to generate antibody subunits. Using the Fab region, as opposed to the full-length antibody, allowed for both higher resolution SEC-MALS experiments to be performed, and supported structural determination of the complexes.
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Localization of O-glycan Sites in CD24Fc by OpeRATOR® Digestion and LC-MS Analysis

November 1, 2022 | Applications, References |

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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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

September 23, 2021 | Applications, References |

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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FabRICATOR® in a DAR Analysis of Antibody-Oligonucleotide Conjugates

September 16, 2021 | Applications, References |

In this paper, scientists at Spectroswiss and Orion Pharma present a novel strategy for DAR analysis of structurally complex antibody-oligonucleotide conjugates using SmartEnzymes™ and a proteoform integration approach combined with native SEC-Orbitrap FTMS.
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