Molecular Biotechnology

, Volume 14, Issue 3, pp 241–249 | Cite as

Analysis of glycoprotein heterogeneity by capillary electrophoresis and mass spectrometry

  • Andrew D. Hooker
  • David C. James

Abstract

Glycosylation is a complex posttranslational modification that can result in extensive heterogeneity for recombinant glycoproteins produced by eukaryotic systems. The carbohydrate moiety of a recombinant glycoprotein may affect the immunogenicity, half-life, bioactivity, and stability of a potential therapeutic product. Regulatory authorities such as the US Food and Drug Administration demand increasingly sophisticated carbohydrate analysis to ensure product characterization, batch-to-batch consistency, and stability.

The advent of new technologies for analysis of biopolymers by capillary electrophoresis and mass spectrometry has revolutionized strategies for recombinant protein characterization. In particular, recent advances in matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry now permit relatively rapid and detaned assessment of glycoprotein and oligosaccharide structure. In this article, we describe some applications of capillary electrophoresis and mass spectrometry to monitor the glycosylation associated with a model recombinant glycoprotein, human interferon-γ.

Index Entries

Glycosylation microheterogeneity recombinant glycoproteins oligosaccharides capillary electrophoresis mass spectrometry 

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Copyright information

© Humana Press Inc 2000

Authors and Affiliations

  • Andrew D. Hooker
    • 1
  • David C. James
    • 2
  1. 1.Oxford GlycoSciences (UK) LtdAbingdonUK
  2. 2.Research School of BiosciencesUniversity of KentCanterburyUK

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