title: Automated, high-throughput IgG-antibody glycoprofiling platform.
abstract: One of today's key challenges is the ability to decode the functions of complex carbohydrates in various biological contexts. To generate high-quality glycomics data in a high-throughput fashion, we developed a robotized and low-cost N-glycan analysis platform for glycoprofiling of immunoglobulin G antibodies (IgG), which are central players of the immune system and of vital importance in the biopharmaceutical industry. The key features include (a) rapid IgG affinity purification and sample concentration, (b) protein denaturation and glycan release on a multiwell filtration device, (c) glycan purification on solid-supported hydrazide, and (d) glycan quantification by ultra performance liquid chromatography. The sample preparation workflow was automated using a robotic liquid-handling workstation, allowing the preparation of 96 samples (or multiples thereof) in 22 h with excellent reproducibility and, thus, should greatly facilitate biomarker discovery and glycosylation monitoring of therapeutic IgGs.
authors.affiliation: NIBRT GlycoScience Group, NIBRT- The National Institute for Bioprocessing Research and Training , Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.
authors.forename: Henning
authors.initials: H
authors.lastname: Stöckmann
authors.forename: Barbara
authors.initials: B
authors.lastname: Adamczyk
authors.forename: Jerrard
authors.initials: J
authors.lastname: Hayes
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000032:analysis*; D057205:Automation, Laboratory / Q000379:methods*; D006031:Glycosylation; D057166:High-Throughput Screening Assays / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G