Abstract
Bonded-phase silica gels were optimized for peptide and protein separations. The optimization involved choice of alkyl chain length and composition, as well as pore sizes of the silica gels. The identities of the alkyl chains were examined by use of solid-state 13C and 29Si cross-polarization magic angle spinning nuclear magnetic resonance spectroscopy (CP/MAS NMR). Furthermore, 13C CP/MAS NMR spectroscopy was used together with elemental analysis for monitoring the synthesis. It was shown that the selectivity for peptides was affected by changing the ligand density. Optimal resolution of proteins was obtained with a low substitution of octyldimethylsilyl chains in combination with ca. 80% of trimethylsilyl chains. The influence of column length and flow-rates on the separation of proteins of similar hydrophobicity, as well as an on/off mechanism in short columns for proteins with a large difference in hydrophobicity, is discussed.
Published Version
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