Glycans play a critical role in the structure, function, and therapeutic efficacy of monoclonal antibodies (mAbs). These carbohydrate chains, which are covalently attached to the antibody’s protein backbone through post-translational modifications, influence a wide range of properties that are essential for both the development and clinical performance of mAb-based therapeutics.
Key Roles of Glycans in Monoclonal Antibodies:
- Stability and Solubility: Glycosylation affects the structural stability and solubility of mAbs.
- Effector Function and Immunogenicity: Glycans will significantly impact the antibody’s ability to interact with immune cells.
- Pharmacokinetics: Glycosylation affects the half-life of monoclonal antibodies.
- Activity and Binding Affinity: Glycans can influence the antibody’s binding affinity to its target antigen.
- Manufacturing Consistency: Consistent and controlled glycosylation patterns are essential to ensure batch-to-batch consistency in the production of monoclonal antibodies.
Aragen provides a highly reproducible and robust released N-linked glycan assay for comprehensive profiling of glycosylation molecule. In this method, glycans are enzymatically cleaved from the antibody, then labelled with a 2-AB/procaine-derived fluorescent dye. After cleaning the labelled glycans, they are separated using LC/LC-MS (see chromatogram graphic).
Our Fluorescence detector, coupled with online ESI mass spectrometry (MS), ensures accurate detection and quantification of various glycan structures. These structures are then categorized based on their structural classification and their monosaccharide composition like; fucose, galactose, high mannose and sialic acid variants.
The chromatogram illustrates the major Glycoforms identified in IgG1 molecule which was produced in the CHO cell line and SP2/O cell line.
