Fast profiling N-glycans in biotherapeutic antibodies by UHPLC-FLD with MS confirmation
Hear Dr. Stefan Mittermayr of the National Institute of Bioprocessing Research and Training (NIBRT) shares the development and optimization of a widely applicable UHPLC approach to fast, comprehensive profiling of 2-AA and 2-AB labeled glycans in IgG antibodies. the approach was validated with a human serum IgG and a commercial chimeric IgG1 mAb (infliximab). Glycan profiles were confirmed by exoglycosidase enzyme digestion and high-resolution, accurate-mass mass spectrometry, allowing increased sample throughput.
An ultrafast, batch-to-batch comparison of monoclonal antibody glycosylation
To develop a high-throughput screening method using HILIC UHPLC separation of 2AA-labelled glycans in a model mAb as a proof of concept for a more general approach to mAb glycoprofiling capable of identifying differences in high abundance gylcoforms. The separation must be rapid but requires sufficient resolution to allow batch-to-batch differences in the glycan profile to be identified and subsequently characterized by high-resolution, accurate-mass mass spectrometry.
Comprehensive protein glycosylation comparison of an innovator monoclonal antibody to a candidate biosimilar by HILIC UHPLC analysis
The goals of this application are to apply a fast, hydrophilic interaction UHPLC approach to the comprehensive glycan profiling of 2-AA labelled candidate biosimilar and an innovator mAb, to determine variations in glycan profile are observable between samples, and to confirm the glycan profiles by exoglycosidase enzyme digestion and high-resolution, accurate-mass (HRAM) mass spectrometry.