Protein glycosylation is essential, as variations in glycan structures can influence the structural stability, biological activity, and therapeutic performance of proteins. The complexity of glycoproteins typically requires advanced analytical approaches for structural determination. Removal of glycans from glycoproteins simplifies the analysis of proteins and enables characterization of glycosylation sites and polypeptide structures. In this study, we evaluated the performance of an immobilized enzyme reactor (IMER) containing peptide N-glycosidase from Rudaea cellulosilytica (PNGase Rc) for online deglycosylation of monomeric intact glycoproteins. We selected human haptoglobin, human transferrin, anti-hHD6, bevacizumab, and trastuzumab as model substrates. The results demonstrate that the PNGase Rc IMER enables rapid and efficient online deglycosylation of glycoproteins and antibodies bearing complex sialylated and core-fucosylated glycans, highlighting its potential in bioanalytical workflows, quality control during biopharmaceutical development, and stability tests.
Molnárová et al. (Mon,) studied this question.