mAb-Drug Potency Determination: Profiling of mAb Glycosylations using FcR–IIIA Affinity Chromatography

Register Free: https://www.chromatographyonline.com/lcgc_w/glycosylations

Event Overview:

Due to the complex nature of monoclonal antibodies, particularly N-linked glycan heterogeneity at the conserved Asn-297 in the Fc region, characterization of mAb-drugs’ critical quality attributes is necessary to maintain the product’s consistency and efficacy. The glycosylation of mAbs can have a positive or negative effect on antibody effector functions. Improved Fc glycoengineering strategies have become necessary to develop and facilitate the next generation of therapeutic mAbs with improved therapeutic potency. The TSKgel® FcR-IIIA analytical and semi-preparative columns separate mAbs into multiple subsets based on their affinity to the FcγR-IIIA ligand which also correlates with their glycan distribution. In this webcast, several novel workflows will be presented that harness the profiling power of these columns to identify N-linked glycan compositions and contents, as well as to determine changes in glycosylation status.

• The analytical TSKgel FcR-IIIA-NPR column provides a quick assessment of mAb glycosylation distribution, either through UV profiling or in combination with high-resolution mass spectrometry detection
• The semi-preparative TSKgel FcR-IIIA-5PW column enables the collection of peaks for orthogonal studies. N-linked glycans from the samples will be analyzed using HILIC-MS and peptide mapping technique on RP-UV-MS
• A novel FcR-based chromatography technique is suitable for mAb-drug potency analysis: to monitor mAb-drug potency from a bioreactor and to monitor N-linked glycosylation remodeling for ADC development

Key Learning Objectives:

• Determine mAb-drug potency using a fast and effective HPLC FcR-IIIA affinity chromatography technique
• Comprehensive profiling of mAb glycoforms using the TSKgel FcR-IIIA affinity columns: from online native mass spectrometry to offline N-linked glycan analysis
• Understand the effects of different glycan distributions on mAbs and their impact on mAb-drug potency

Who Should Attend:

• Analytical scientists, method developers and quality control personnel working in the biopharmaceutical industry

Speakers

Heidi Vitrac, Ph.D.
Applications Scientist
Tosoh Bioscience LLC

Heidi’s academic career in France and in the US has centered around membrane biology and lipid-protein interactions. This work has been in four main productive phases: Her PhD work on the mechanisms of oxidation of biomolecules upon exposure to oxidative stress (2001-2005) was at the University of Paris Descartes. This was followed by post-doctoral research projects on the mechanisms of protein folding at the membrane interface (2006-2007) in Grenoble, France, and continued with the role of lipid-protein interactions in the structure and function of integral membrane proteins at the University of Texas Health Science Center, Houston (2008-2013). As an Assistant Professor at the University of Texas (2013-2020), her research focused on the crosstalk between membrane phospholipid metabolism, bacterial physiology and stress adaptation. Throughout her research career, she adopted multidisciplinary analytical, biophysical, and biochemical approaches, including multiple modes of chromatography and mass spectrometry (proteomics, lipidomics and metabolomics) to understand the mechanisms of lipid-dependent defects in the development and progression of diseases.

Heidi joined Tosoh Bioscience LLC in December 2020 as an Applications Scientist, where she is developing analytical chromatography applications on TSKgel columns for the online MS characterization of monoclonal antibodies and their glycosylation profile.

Register Free: https://www.chromatographyonline.com/lcgc_w/glycosylations