Chromatography glycoprotein analysis

The use of ConA affinity chromatography in SDS has proved to be very useful for glycoprotein analysis because it combines complete dissociation of the glycoproteins from associated components with a rapid, specific, and high yield method of isolation. Most importantly, the glycoproteins reform the complex after displacement of SDS by nonionic detergent (Bando et al., 1996 Li et al., 1999). 

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

Lampio, A. and Finne, J., Sugar analysis of glycoproteins and glycolipids after methanolysis by high-performance liquid chromatography with pulsed amperometric detection, Anal. Biochem., 197, 132, 1991. 

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. 

Lochnit, G. and Geyer, R., An optimized protocol for nano-LC-M ALDI-TOE-MS coupling for the analysis of proteolytic digests of glycoproteins. Biomedical Chromatography 18(10), 841-848, 2004. 

Imre, T., Schlosser, G., Pocsfalvi, G., Sicihano, R., Mobiar-SzoUosi, E., Kremmer, T., Malorni, A., and Vekey, K., Glycosylation site analysis of human alpha-l-acid glycoprotein (AGP) by capillary liquid chromatography-electrospray mass spectrometry. Journal of Mass Spectrometry 40(11), 1472-1483, 2005. 

K. A. Thomsson, N. G. Karlsson, and G. C. Hansson, Liquid chromatography-electrospray mass spectrometry as a tool for the analysis of sulfated oligosaccharides from mucin glycoproteins, J. Chromatogr. A, 854 (1999) 131-139. 

N. Kawasaki, M. Ohta, S. Hyuga, O. Hashimoto, and T. Hayakawa, Analysis of carbohydrate heterogeneity in a glycoprotein using liquid chromatography/mass spectrometry and liquid chromatography with tandem mass spectrometry, Anal. Biochem., 269 (1999) 297-303. 

M. Chaplin, A rapid and sensitive method for the analysis of carbohydrate compounds in glycoproteins using gas-liquid chromatography, Anal. Biochem., 723(2) 336-341 (1985). 

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]. 

A water-soluble, 50%-methanol-soluble, acidic glycoprotein was isolated from rat brain by affinity chromatography on con A-Sepharose.855 The glycoprotein, pure by poly(acrylamide) gel-electrophoresis at pH 8.8, had an apparent molecular weight of 14,500 1,400 by dodecyl sodium sulfate gel-electrophoresis. No analysis for carbohydrate was reported.855... 

It is well known that drugs bind to plasma proteins, particularly to serum albumin and a-acid glycoprotein, and that only the unbound, or free, fraction is responsible for any pharmacological effect. For protein-drug binding studies size-exclusion chromatography in one of three variants—namely, the Hum-mel-Dreyer method (1962), the vacancy peak method (Sebille, et al., 1979), and frontal analysis (Cooper and Wood, 1968)—is the traditional method of... 

---