title: Rapid quantitation of monoclonal antibody N-glyco-occupancy and afucosylation using mass spectrometry.
abstract: N-glyco-occupancy and afucoslyation level are two important quality attributes associated with N-glycosylation of therapeutic monoclonal antibodies (mAbs). We report here a fast mass spectrometry-based workflow for quantification of N-glycan site-occupancy and afucoslyation level of mAbs with improved throughput, precision, sensitivity and robustness. This method uses the deglycosylation after the first GlcNAc and inter-chain reduction of the mAbs, followed by liquid chromatography/mass spectrometry (LC-MS) analysis. The entire process can be completed within one hour, which provides a rapid quantitation of N-glyco-occupancy and afucosylation to support high-throughput cell line selection and process development for mAb biopharmaceuticals.
authors.affiliation: Analytical Development, Biogen, United states.
authors.forename: Suli
authors.initials: S
authors.lastname: Liu
authors.affiliation: Analytical Development, Biogen, United states. Electronic address: li.zang@biogen.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zang
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry*; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry*; D005643:Fucose / Q000737:chemistry*; D006031:Glycosylation; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D005643:Fucose; D000117:Acetylglucosamine
keywords: Afucosylation; Glyco-occupancy; High-throughput; Liquid chromatography/mass spectrometry; Monoclonal antibody