Capillary electrophoresis in the N-glycosylation analysis of biopharmaceuticals

•Comprehensive N-glycosylation analysis techniques for biotherapeutics are described.•Identification of potentially immunogenic epitopes is very important.•Sample-preparation issues are discussed, including glycan release fluorophore labeling, sample cleanup and capillary electrophoresis analysis.•Carbohydrate sequencing reveals linkage and positional information.•Glycan analysis needs are different in clone selection, product development and lot release.

In N-glycosylation analysis of biopharmaceuticals, analytical need depends on the phase of the manufacturing process. All important glycoanalysis steps are thoroughly discussed. Carbohydrate sequencing by exoglycosidase arrays is described in conjunction with capillary electrophoresis (CE) to identify linkage and positional isomers. A possible automated workflow for N-glycosylation based on CE is outlined.