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Updated: May 12, 2026
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A Quantitative Glycomics and Proteomics Combined Purification Strategy
Published on: March 8, 2016
Online Deglycosylation of Monomeric Intact Proteins Using the PNGase Rc Immobilized-Enzyme Reactor
Katarína Molnárová1, Petr Novák2, Jana Nováková1
1AffiPro s.r.o., Nad Safinou II 365, Vestec 252 50, Czech Republic.
ACS Omega
|May 11, 2026
Summary
This study introduces an immobilized enzyme reactor with peptide N-glycosidase (PNGase Rc) for rapid protein deglycosylation. This method efficiently removes complex glycans from glycoproteins and antibodies, aiding bioanalysis and biopharmaceutical development.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biotechnology
Background:
- Protein glycosylation critically impacts protein structure, activity, and therapeutic efficacy.
- Analyzing complex glycoproteins necessitates advanced techniques for structural elucidation.
- Glycan removal simplifies protein analysis, enabling characterization of glycosylation sites and polypeptide structures.
Purpose of the Study:
- To evaluate an immobilized enzyme reactor (IMER) with peptide N-glycosidase from Rudaea cellulosilytica (PNGase Rc).
- To assess the IMER's performance for online deglycosylation of monomeric intact glycoproteins.
- To demonstrate the utility of PNGase Rc IMER in bioanalytical workflows.
Main Methods:
- Development and testing of an immobilized enzyme reactor (IMER) utilizing peptide N-glycosidase (PNGase Rc).
- Utilized model substrates including human haptoglobin, human transferrin, anti-hHD6, bevacizumab, and trastuzumab.
- Performed online deglycosylation experiments with the PNGase Rc IMER.
Main Results:
- The PNGase Rc IMER demonstrated rapid and efficient online deglycosylation of various glycoproteins and antibodies.
- Successfully removed complex sialylated and core-fucosylated glycans.
- The reactor proved effective for diverse protein substrates.
Conclusions:
- The PNGase Rc IMER is a valuable tool for efficient online deglycosylation of glycoproteins and antibodies.
- This technology holds significant potential for bioanalytical workflows, quality control in biopharmaceutical development, and protein stability testing.
