Work

Online Deglycosylation of Monomeric Intact Proteins Using the PNGase Rc Immobilized-Enzyme Reactor

Year: 2026

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide 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.

Source: ACS Omega

DOI: 10.1021/acsomega.6c00577

Authors: Katarína Molnárová, Petr Novák, Jana Nováková, Petr Pompach

Institution: Czech Academy of Sciences, Institute of Microbiology

Language: English

Topic: Glycosylation and Glycoproteins Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,935

Funders: European Commission, Ministerstvo Školství, Mládeže a Tělovýchovy, Akademie Věd České Republiky, Mikrobiologický Ústav, Akademie Věd České Republiky