title: State-of-the-art technologies for rapid and high-throughput sample preparation and analysis of N-glycans from antibodies.
abstract: Glycosylation is a PTM that occurs during production of many protein-based biologic drugs and can have a profound impact on their biological, clinical, and pharmacological properties. Quality by design, process optimization, and advance in manufacturing technology create a demand for robust, sensitive, and accurate profiling and quantification of antibody glycosylation. Potential drawbacks in antibody glycosylation profiling include the high hands-on time required for sample preparation and several hours for data acquisition and analysis. Rapid and high-throughput (HTP) N-glycan profiling and characterization along with automation for sample preparation and analysis are essential for extensive antibody glycosylation analysis due to the substantial improvement of turnaround time. The first part of this review article will focus on the recent progress in rapid and HTP sample preparation and analysis of antibody glycosylation. Subsequently, the article will cover a brief overview of various separation and mass spectrometric methods for the rapid and HTP analysis of N-glycans in antibodies. Finally, we will discuss the recent developments in process analytical technologies for the screening and quantification of N-glycans in antibodies.
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, PA, USA.
authors.forename: Udayanath
authors.initials: U
authors.lastname: Aich
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, PA, USA.
authors.forename: Jude
authors.initials: J
authors.lastname: Lakbub
authors.affiliation: Biopharmaceutical Analytical Sciences, Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, PA, USA.
authors.forename: Aston
authors.initials: A
authors.lastname: Liu
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D006031:Glycosylation; D057166:High-Throughput Screening Assays / Q000639:trends*; D006801:Humans; D011134:Polysaccharides / Q000032:analysis*
chemical_list: D000911:Antibodies, Monoclonal; D011134:Polysaccharides
keywords: Antibody N-glycosylation; High throughput; MS; Process analytical technologies; Separation