Current Approaches to Glycoprotein Analysis
Andrew K. Hagan, Maomao Wang and Li Liu
Affiliation: Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, P.R. China.
Keywords: Analytical techniques, chemical derivatization, glycosylation, HPLC, mass spectrometry, proteases, therapeutic
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.
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