N-linked glycosylation is a common post-translational modification that imparts structural heterogeneity to recombinant monoclonal antibody therapeutics. The various oligosaccharides attached to the CH2 domains of IgG can impact the efficacy, safety and pharmacokinetics of the product. Depending on the mechanism of action of the biotherapeutic, specific glycan moieties may be deemed critical quality attributes (CQAs) and require appropriate process control and analytical strategies to ensure product quality consistency. Glycan analysis presents a significant challenge because the N-glycosylation biosynthetic pathway results in a heterogeneous population of complex branched structures at each glycosylation site. Consequently, extensive sample handling is required for analysis. At various stages of developing and manufacturing a mAb several levels of glycan analysis are required: (i) full detailed analysis including monosaccharide sequence and linkage of the glycans which usually requires orthogonal technologies (ii) characterisation of the glycans by type to provide particular CQAs (iii) profiling, which can provide a quick comparative overview of the glycosylation.. Glycoanalytical standards are of critical importance in evaluating the suitability of methods intended to monitor glycosylation through the lifecycle of the product. The intact NIST mAb reference material affords an additional supplement to in-house reference material for system suitability, operator training, and analytical method evaluation. For recombinant monoclonal antibody biotherapeutics expressed in mammalian cell culture, the collection of nested N-linked glycan species is relatively restricted, albeit the relative proportions of the glycan species may differ significantly. This consistent population of glycan species justifies the development of a single platform glycan analysis strategy to support a mAb biotherapeutic pipeline. The NIST mAb glycan population was analysed using s
State-of-the-Art and Emerging Technologies for Therapeutic Monoclonal Antibody Characterization
and Mouchahoir, C.
Glycan Analysis of NIST mAb Reference Material, State-of-the-Art and Emerging Technologies for Therapeutic Monoclonal Antibody Characterization, ACS, Washington DC, DC, [online], https://doi.org/10.1021/bk-2015-1201.ch004
(Accessed July 7, 2022)