Simple and Robust N-Glycan Analysis Based on Improved 2-Aminobenzoic Acid Labeling for Recombinant Therapeutic Glycoproteins

Cited 1 time in webofscience Cited 0 time in scopus
  • Hit : 87
  • Download : 0
N-glycans of therapeutic glycoproteins are critical quality attributes that should be monitored throughout all stages of biopharmaceutical development. To reduce both the time for sample preparation and the variations in analytical results, we have developed an N-glycan analysis method that includes improved 2-aminobenzoic acid (2-AA) labeling to easily remove deglycosylated proteins. Using this analytical method, 15 major 2-AA-labeled N-glycans of Enbrel (R) were separated into single peaks in hydrophilic interaction chromatography mode and therefore could be quantitated. 2-AA-labeled N-glycans were also highly compatible with in-line quadrupole time-of-flight mass spectrometry (MS) for structural identification. The structures of 15 major and 18 minor N-glycans were identified from their mass values determined by quadrupole time-of-flight MS. Furthermore, the structures of 14 major N-glycans were confirmed by interpreting the MS/MS data of each N-glycan. This analytical method was also successfully applied to neutral N-glycans of Humira (R) and highly sialylated N-glycans of NESP (R). Furthermore, the analysis data of Enbrel (R) that were accumulated for 2.5 years demonstrated the high-level consistency of this analytical method. Taken together, the results show that a wide repertoire of N-glycans of therapeutic glycoproteins can be analyzed with high efficiency and consistency using the improved 2-AA labeling-based N-glycan analysis method. (C) 2018 American Pharmacists Association (R). Published by Elsevier Inc. All rights reserved.
Publisher
ELSEVIER SCIENCE INC
Issue Date
2018-07
Language
English
Article Type
Article
Keywords

ANION-EXCHANGE CHROMATOGRAPHY; GLYCOSYLATION PROFILES; ANTHRANILIC ACID; LIQUID-CHROMATOGRAPHY; COMPLEX GLYCOPROTEIN; HUMAN-ERYTHROPOIETIN; FLUORESCENCE; ACETYLGLUCOSAMINE; OLIGOSACCHARIDES; DERIVATIZATION

Citation

JOURNAL OF PHARMACEUTICAL SCIENCES, v.107, no.7, pp.1831 - 1841

ISSN
0022-3549
DOI
10.1016/j.xphs.2018.03.013
URI
http://hdl.handle.net/10203/245424
Appears in Collection
BS-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡ Click to see webofscience_button
⊙ Cited 1 items in WoS Click to see citing articles in records_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0