Effects of the redox state of porous graphitic carbon on the retention of oligosaccharides.
Retention of hydrophilic compounds on porous graphitic carbon (PGC) is afforded by polar interactions with induced dipoles within this polarizable stationary phase. These interactions depend on the redox state of PGC, which can be influenced by application of an electrical field or by chemical means. We explored the impact of oxidizing and reducing agents on the retention of fluorescence labeled neutral oligosaccharides. Malto-oligosaccharides were employed as simple model system. Subsequently, the effects on the retention of glycans typical for immunoglobulin G (IgG) antibodies were investigated. Chemical oxidation of the PGC surface increased the retention of all analytes tested. Selectivities were significantly altered by the redox treatment, emphasizing the need for controlling the redox state of PGC to achieve reproducible conditions. Furthermore a column pre-conditioning protocol is presented, which allowed for reproducible chromatography of neutral IgG glycans.
Melmer
Michael
M
Sandoz GmbH, Biochemistrasse 10, 6250 Kundl, Austria.
Stangler
Thomas
T
Premstaller
Andreas
A
Lindner
Wolfgang
W
D002853:Chromatography, Liquid / Q000379:methods*; D006108:Graphite / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D008320:Maltose / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D016062:Porosity; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
D007074:Immunoglobulin G; D009844:Oligosaccharides; D008320:Maltose; D006108:Graphite