High-pH anion-exchange chromatography

Oligosaccharides were isolated preparatively by high-pH anion exchange chromatography carried out on a LC-system (Dionex Corporation, Sunnyvale CA) equipped with a CarboPac PA-1 column (9 x 250 mm), coupled to a Spectra System AS 3500 auto sampler. The detection was carried out using Pulsed Amperometric Detection (PAD-II). 

Townsend, R. R., Atkinson, R H., and Trimble, R. B., Separation of high-mannose isomers from yeast and mammalian sources using high pH anion-exchange chromatography, Carb. Res., 215, 211. 1991. 

Kotani, N. and Takasaki, S., Analysis of 2-aminobenzamide-labeled oligosaccharides by high-pH anion-exchange chromatography with fluorometric detection, Anal. Biochem., 264, 66, 1998. 

High-pH anion-exchange chromatography (HPAEC) of monosaccharides and oligosaccharides has been more recently developed as a useful analytical tool. At sufficiently alkaline pH (12 to 14)... 

Hardy, M. R., and Townsend, R. R. (1994). High-pH Anion-Exchange Chromatography of Glycoprotein-Derived Carbohydrates. Methods Enzymol 230 208. 

After isolation and identification of the peptides containing these glycosylation sites, high pH anion exchange chromatography, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, and electrospray ionization mass spectrometry (ES-MS) were utilized to evaluate carbohydrate microheterogeneity. 

Table 1 summarizes the carbohydrate structural data deduced from the three detection methodologies, MALDI-TOF, ES-MS, and high pH anion exchange chromatography. The N-glycosylation site proved to be occupied with several structures. The glycopeptides were partially separated as five peptides, at retention times of 19.4, 20.3, 20.7, 20.9, and 21.3 minutes, and all of these glycopeptides contained the same amino acid sequence, N-terminal amino acids 1-17. 

Characterization of glycoproteins requires application of several chromatographic and mass spectrometric analytical techmques. High pH anion exchange chromatography of oligosaccharides and asialo oligosaccharides was... 

Townsend RR, Basa LJ, Spellman MW (1996) Identification and characterization of glycopeptides in tryptic maps by high-pH anion exchange chromatography. In Karger BL,Hancock (eds) Methods enzymol,vol 271. Academic Press, NY, p 135... 

J. Heinrich and P. Mischnick, A rapid method for the determination of the substitution pattern of O-mefliylated 1,4-glucans by high-pH anion-exchange chromatography wifli pulsed amperometric detection, /. Chromatogr. A, 749 (1996)41 5. 

Hermentin, P. et al. The mapping by high-pH anion-exchange chromatography with pulsed amperometric detection and capillary electrophoresis of the carbohydrate moieties of human plasma alpha 1-acid glycoprotein. Anal Biochem, 206, 419,1992. 

High pH anion exchange chromatography (HPAEC) has been applied to the separation of neutral V-linked oligosaccharides, oligosaccharides derivatized by p-nitrobenzylhydroxylamine and various D-mannose derivatives. In the last case the relationship between retention times and structure was examined. ... 

Postreleased N- or O-linked glycans may be cleaned up, and directly analyzed on a pellicular anion resin column in strong alkaline conditions (e.g., 0.1 M NaOH), a method commonly known as high pH anion-exchange chromatography... 

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