Ultrasensitive capillary electrophoretic analysis of potentially immunogenic carbohydrate residues in biologics

Galactose-α-1,3-galactose containing oligosaccharides

Zoltan Szabo, A. Guttman, Jonathan Bones, Randi L. Shand, David Meh, Barry L. Karger

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

With the recent growth of the global monoclonal antibody market, ultrasensitive techniques are required for rapid analysis of possible immunogenic residues, such as galactose-α-1,3-galactose (α-1,3-Gal) on therapeutic proteins expressed in murine or CHO cell lines. We report a capillary electrophoretic approach in conjunction with exoglycosidase digestion for structural elucidation of IgG N-glycans containing the above immunogenic epitope. The method uses commercially available reagents and instrumentation, thus making it readily available for implementation and validation within the biotechnology industry. The method was first evaluated using polyclonal mouse IgG N-glycans which are known to contain α-1,3-Gal containing epitopes. High reproducibility in migration time enabled determination of GU values for five α-1,3-Gal containing structures when the data from all samples analyzed was combined. The method was successfully applied to the analysis of a NCI reference standard monoclonal antibody and two development phase monoclonal antibodies. The limit of detection and limit of quantitation were 1 and 2 μg of intact protein IgG starting material, respectively, further indicating the high sensitivity of the described method.

Original languageEnglish
Pages (from-to)1612-1619
Number of pages8
JournalMolecular Pharmaceutics
Volume9
Issue number6
DOIs
Publication statusPublished - Jun 4 2012

Fingerprint

Biological Products
Oligosaccharides
Galactose
Carbohydrates
Immunoglobulin G
Monoclonal Antibodies
Polysaccharides
Epitopes
CHO Cells
Glycoside Hydrolases
Biotechnology
Limit of Detection
Digestion
Industry
Proteins
Cell Line
Growth
Therapeutics

Keywords

  • alpha galactose
  • capillary electrophoresis
  • exoglycosidase digestion
  • glycomics
  • immunogenic epitopes
  • monoclonal antibody
  • therapeutic antibody

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Molecular Medicine
  • Drug Discovery

Cite this

Ultrasensitive capillary electrophoretic analysis of potentially immunogenic carbohydrate residues in biologics : Galactose-α-1,3-galactose containing oligosaccharides. / Szabo, Zoltan; Guttman, A.; Bones, Jonathan; Shand, Randi L.; Meh, David; Karger, Barry L.

In: Molecular Pharmaceutics, Vol. 9, No. 6, 04.06.2012, p. 1612-1619.

Research output: Contribution to journalArticle

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