Development of a high performance anion exchange chromatography analysis for mapping of oligosaccharides

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jul 1;877(20-21):1827-32. doi: 10.1016/j.jchromb.2009.05.003. Epub 2009 May 13.

Abstract

In the present study a HPAEC-PAD method is described that was developed for monitoring the consistency of N-glycosylation during the production and purification of recombinant proteins and monoclonal antibodies. The method successfully separated 18 neutral and sialylated oligosaccharides. Results obtained were compared with MALDI-TOF MS and it was shown that both methods gave similar results. In addition, a method validation was performed showing that the HPAEC-PAD analysis was well suited for the mapping and characterization of oligosaccharides. The method was found to be robust and additionally the precision was significantly better compared to the MALDI-TOF MS method.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry*
  • CHO Cells
  • Chromatography, Ion Exchange / methods*
  • Cricetinae
  • Cricetulus
  • Glycosylation
  • Oligosaccharides / analysis*
  • Oligosaccharides / chemistry
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Antibodies, Monoclonal
  • Oligosaccharides
  • Recombinant Proteins