title: Current approaches to glycoprotein analysis.
abstract: Glycoproteins are becoming increasingly relevant in therapeutics, including tissue plasminogen activator for the treatment of myocardial infarction and strokes, erythropoietin for anemia and various monoclonal antibody-based treatments for cancer. Protein N- and O-glycosylation is perhaps the most crucial and immensely complex posttranslational modification that proteins undergo, and its characterization presents a major challenge. This review will discuss current techniques for the characterization of glycoproteins, with a focus on therapeutic glycoproteins where available. The crucial analytical techniques, such as high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), and mass spectrometry (MS) will be described, alongside the necessary chemical labeling methods for sensitive detection. The well-established chemical and enzymatic methods for oligosaccharide release from proteins will be discussed, as will more modern methods based on exhaustive protein hydrolysis with non-specific proteases.
authors: Andrew K
authors: AK
authors: Hagan
authors: Maomao
authors: M
authors: Wang
authors: Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, P.R. China. lichen.liu@njau.edu.cn.
authors: Li
authors: L
authors: Liu
mesh_terms: D000970:Antineoplastic Agents / Q000032:analysis* / Q000737:chemistry; D002240:Carbohydrate Sequence; D002317:Cardiovascular Agents / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D006397:Hematinics / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000970:Antineoplastic Agents; D002317:Cardiovascular Agents; D005456:Fluorescent Dyes; D006023:Glycoproteins; D006397:Hematinics; D011134:Polysaccharides