title: Evaluation of oligosaccharide methods for carbohydrate analysis in a fully human monoclonal antibody and comparison of the results to the monosaccharide composition determination by a novel calculation.
abstract: Carbohydrates can change a drug's properties including solubility, affinity towards antigen, pharmacokinetics and pharmacodynamics. Due to this importance, carbohydrate composition is utilized as a parameter to evaluate a drug candidate's quality. In this study, the compositional monosaccharides of a drug candidate are measured by HPAEC-PAD, while the oligosaccharides are studied by HPAEC-PAD, CE-LIF and LC-MS. The advantages and limitations of these various approaches for oligosaccharide analysis are reviewed in this work. While the methods used for oligosaccharide analysis are well established we have devised a new and novel calculation for determining monosaccharide content using the relative percentages of the N-glycans. This calculation was used to evaluate the accuracy of the oligosaccharide determination methods by comparison of the N-glycan data to the experimental monosaccharide data. The results obtained from this novel calculation demonstrate that the relative abundance of carbohydrates as determined from these various approaches are consistent.
authors.affiliation: Department: Analytical Development, Medarex, Inc., Bloomsbury, NJ 08804, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Adamo
authors.forename: Difei
authors.initials: D
authors.lastname: Qiu
authors.forename: Lawrence W
authors.initials: LW
authors.lastname: Dick
authors.forename: Ming
authors.initials: M
authors.lastname: Zeng
authors.forename: An-Horng
authors.initials: AH
authors.lastname: Lee
authors.forename: Kuang-Chuan
authors.initials: KC
authors.lastname: Cheng
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000737:chemistry; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis* / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D055641:Mathematical Concepts; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis* / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D009005:Monosaccharides; D009844:Oligosaccharides