Capillary electrophoresis glycoproteins analysis

Landers, J. P., Oda, R. P., Madden, B. J., and Spelsberg, T. C., High-performance capillary electrophoresis of glycoproteins the use of modifiers of electro-osmotic flow for analysis of microheterogeneity, Anal. Biochem., 205, 115, 1992. 

Choudhary, G., Chakel, J., Hancock, W., Torres-Duarte, A., McMahon, G., and Wainer, I., Investigation of the potential of capillary electrophoresis with offline matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for clinical analysis examination of a glycoprotein factor associated with cancer cachexia, Anal. Chem. 71, 855, 1999. 

Kakehi, K., Kinoshita, M., and Nakano, M. 2002. Analysis of glycoproteins and the oligosaccharides thereof by high performance capillary electrophoresis - significance in regulatory studies on biopharmaceutical products. Biomedical Chromatography 16(2), 103-115. 

Kakehi, K., and Honda, S. (1996). Analysis of glycoproteins, glycopeptides and glycoprotein-derived oligosaccharides by high-performance capillary electrophoresis.. Chromatogr. A 720, 377-393. 

Kamoda, S., and Kakehi, K. (2006). Capillary electrophoresis for the analysis of glycoprotein pharmaceuticals. Electrophoresis 27, 2495—2504. 

Taverna, M., Baillet, A., Biou, D., Schluter, M., Werner, R., and Terrier, D. (1992). Analysis of carbohydrate-mediated heterogeneity and characterization of N-linked oligosaccharides of glycoproteins by high-performance capillary electrophoresis. Electrophoresis 13, 359—366. 

Yeung, B., Porter, T. J., and Vath, J. E. (1997). Direct isoform analysis of high-mannose containing glycoproteins by on-line capillary electrophoresis electrospray mass spectrometry. Anal. Chem. 69, 2510-2516. 

Chakel, J. A., Erno, J., Hancock, W. S., and Swedberg, S. A. (1997). Analysis of recombinant DNA-derived glycoproteins via high-performance capillary electrophoresis coupled with off-line matrix-assisted laser desorption ionization time-of-fiight mass spectrometry. /. Chromatogr. B 689, 215—220. 

Electrospray mass spectrometry has developed into a well-established method of wide scope and potential over the past 15 years. The softness of electrospray ionization has made this technique an indispensable tool for biochemical and biomedical research. Electrospray ionization has revolutionized the analysis of labile biopolymers, with applications ranging from the analysis of DNA, RNA, oligonucleotides, proteins as well as glycoproteins to carbohydrates, lipids, gly-colipids, and lipopolysaccharides, often in combination with state-of-the-art separation techniques like liquid chromatography or capillary electrophoresis [1,2]. Beyond mere analytical applications, electrospray ionization mass spectrometry (ESMS) has proven to be a powerful tool for collision-induced dissociation (CID) and multiple-stage mass spectrometric (MSn) analysis, and - beyond the elucidation of primary structures - even for the study of noncovalent macromolecular complexes [3]. 

While the discussion in this text focusses on the use of LC-MS, on-line capillary electrophoresis-MS is another important tool in glycopeptide and glycoprotein analysis, as shown by the data reviewed by Zamfir and Peter-Katahnic [89]. 

Capillary isoelectric focusing (CIEF) is a high-resolution technique for protein and peptide separation performed at academic sites and in the biotechnology and pharmaceutical industries for the analysis and characterization of, for example, recombinant antibodies and other recombinant proteins, isoforms of glycoproteins, point mutations in hemoglobin, and peptide mapping. Also, hyphenation to mass spectrometry and chip-based CIEF (microfabrication) have shown promise. CIEF kits and specific recipes/application notes are available from vendors of capillary electrophoresis (CE) equipment, as are a vast amount of publications and handbooks of CE published over recent years. 

Carbohydrates play an important role in many research and industrial domains. The huge number of stereoisomers, the immense combination possibilities of carbohydrate monomers in oligosaccharides, and the lack of chromophores are the major problems in the analysis of carbohydrates. Capillary electrophoresis (CE), in its various modes of operation, has been developed as a very useful tool in the analysis of carbohydrate species such as monosaccharides and oligosaccharides, glycoproteins, and glycopeptides. 

Balaguer, E. and Neusuess, C. (2006) Glycoprotein characterization combining intact protein and glycan analysis by capillary electrophoresis-electrospray ionization-mass spectrometry. Anal Chem, 78 (15), 5384-5393. 

Kinoshita M, Mnrakami E, Oda Y, et al. (2000). Comparative stndies on the analysis of glycosylation heterogeneity of sialic acid-containing glycoproteins nsing capillary electrophoresis. J. Chromatog. A. 866 261-271. 

Methods for Glycoprotein Analysis Using Liquid Chromatography and Capillary Electrophoresis/Electrospray Mass Spectrometry... 

---