In this study, several glycoproteins: bovine α1-acid glycoprotein, fetuin, and human α1-acid glycoprotein, were analyzed using nanoLC-MS/MS. The performance of three different stationary phases – C8, C18, and porous graphite – was systematically evaluated and optimized for glycopeptide separation. A porous graphite column was chosen as the optimum stationary phase for glycoprotein analysis. An LTQ Orbitrap XL hybrid mass spectrometer equipped with electron transfer dissociation (ETD) was used to acquire, within a single analysis, high-resolution, high-mass-accuracy full MS spectra, collision-induced dissociation (CID) MS/MS spectra for glycan composition analysis, and ETD MS/MS spectra for peptide structure information. As a result, characterization of glycopeptides was achieved within a single LC-MS/MS run.
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We thank David Fisher at Thermo Fisher Scientific for editing and proof reading of this manuscript.
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