Antibody Glycans Characterization

Glycosylation is one of the main IgG post-translational modifications and has essential roles in antibody effectors functions, immunogenicity and plasmatic clearance. In this chapter we discuss and provide detailed protocols for IgG homogeneity and determination of level of glycosylation by Capillary Electrophoresis-Sodium Dodecyl Sulfate (CE-SDS), for orthogonal glyco-profiling of IgG-released glycans by Capillary Electrophoresis with a Laser-Induced Fluorescent Detector (CE-LIF) and by Normal-Phase High Performance Liquid Chromatography (NP-HPLC), as well as glycans fine structure assessment by Matrix-Assisted Laser Desorption/ Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) and by nano-Liquid Chromatography tandem mass spectrometry fragmentation (nano-LC-MS/MS).