Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs

Nan Yang, Elisha Goonatilleke, Dayoung Park, Ting Song, Guorong Fan, Carlito B Lebrilla

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). Our results showed that irrespective of the IgG subclass expressed in the drugs, the N-glycopeptide profiles are nearly the same but differ in abundances. In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. Such multilevel structural characterization is important for the successful development of therapeutic antibodies.

Original languageEnglish (US)
Pages (from-to)7091-7100
Number of pages10
JournalAnalytical Chemistry
Volume88
Issue number14
DOIs
StatePublished - Jul 19 2016

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Glycosylation
Polysaccharides
Glycopeptides
Immunoglobulin G
Monoclonal Antibodies
Monitoring
Pharmaceutical Preparations
Antibodies
Liquid chromatography
Complex Mixtures
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. / Yang, Nan; Goonatilleke, Elisha; Park, Dayoung; Song, Ting; Fan, Guorong; Lebrilla, Carlito B.

In: Analytical Chemistry, Vol. 88, No. 14, 19.07.2016, p. 7091-7100.

Research output: Contribution to journalArticle

Yang, N, Goonatilleke, E, Park, D, Song, T, Fan, G & Lebrilla, CB 2016, 'Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs', Analytical Chemistry, vol. 88, no. 14, pp. 7091-7100. https://doi.org/10.1021/acs.analchem.6b00963
Yang, Nan ; Goonatilleke, Elisha ; Park, Dayoung ; Song, Ting ; Fan, Guorong ; Lebrilla, Carlito B. / Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. In: Analytical Chemistry. 2016 ; Vol. 88, No. 14. pp. 7091-7100.
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