Glycosylation is one of the key attributes to monitor during the development and production of a biotherapeutic protein. This is due to the wide range of physicochemical characteristics that are influenced by glycans. These include monoclonal antibody (mAb) stability, antibody dependent cell-mediated cytotoxicity (ADCC), the serum half-life of the drug, and some non-human glycan epitopes present on recombinant biotheraputics can elicit an immunogenic response. Because of the importance of glycosylation, regulatory agencies now require in-depth characterization using state-of-the-art analytical methods. However, glycans are challenging to analyze by LC-MS methods as they are not optically active nor do they possess a basic site needed for positive mode mass spectral ionization in their native state. Traditionally, sample preparation protocols have been laborious, oftentimes tedious, and time consuming. Further compounding the challenges of glycan analysis has been the lack of widely available bioinformatics tools to quickly and effectively process LC-MS data.
In this GEN webinar, William Alley, Ph.D., Senior Scientist for Waters, will describe a streamlined glycan analysis platform beginning with sample preparation and the ensuing data collection and processing, including a library search based on Glucose Units (GU) and accurate mass for structural identification.
Dr. Alley will discuss a high throughput, simplified sample preparation procedure using the GlycoWorks™ RapiFluor-MS™ N-Glycan Kit which facilitates sensitive glycan measurements for both FLR and MS. He will further describe the RFMS Glycan GU Scientific Library, jointly developed with the National Institute for Bioprocessing, Research and Training (NIBRT) in Ireland, for structural identification of the collected data to simplify this task and increase the overall throughput of data analysis. He will conclude with a comparative case study of the glycan profiles of innovator and biosimilar infliximab using this platform.