Enzymatic removal of N-glycans by PNGase F coated magnetic microparticles

Judit Bodnar, Akos Szekrenyes, Marton Szigeti, Gabor Jarvas, Jana Krenkova, Frantisek Foret, A. Guttman

Research output: Contribution to journalArticle

10 Citations (Scopus)


Investigation of protein glycosylation is an important area in biomarker discovery and biopharmaceutical research. Alterations in protein N-glycosylation can be an indication of changes in pathological conditions in the medical field or production parameters of biotherapeutics. Rapid development of these disciplines calls for fast, high-throughput, and reproducible methods to analyze protein N-glycosylation. Currently used methods require either long deglycosylation times or large excess of enzymes. In this paper, we report on the use of PNGase F immobilization onto the surface of magnetic microparticles and their use in rapid and efficient removal of N-glycans from glycoproteins. The use of immobilized PNGase F also allowed reusability of the enzyme-coated beads as the magnetic microparticles can be readily partitioned from the sample by a magnet after each deglycosylation reaction. The efficiency and activity of the PNGase F coated magnetic beads was compared with in-solution enzyme reactions using standard glycoproteins possessing the major N-glycan types of neutral, high mannose, and highly sialylated carbohydrates. The PNGase F coated magnetic beads offered comparable deglycosylation level to the conventional in-solution based method in 10-min reaction times for the model glycoproteins of immunoglobulin G (mostly neutral carbohydrates), ribonuclease B (high mannose type sugars), and fetuin (highly sialylated oligosaccharides) with the special features of easy removal of the enzyme from the reaction mixture and reusability.

Original languageEnglish
Publication statusAccepted/In press - 2016



  • Deglycosylation
  • Enzyme immobilization
  • Magnetic microparticles
  • PNGase F

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Bodnar, J., Szekrenyes, A., Szigeti, M., Jarvas, G., Krenkova, J., Foret, F., & Guttman, A. (Accepted/In press). Enzymatic removal of N-glycans by PNGase F coated magnetic microparticles. Electrophoresis. https://doi.org/10.1002/elps.201500575