使用多空石墨色谱柱和LTQ Orbitrap XL ETD组合质谱分析糖基化肽段

简介：

In this study, several glycoproteins: bovine α1-acid glycoprotein, fetuin and human α1-acid glycoprotein, were analyzed using nano LC-MS/MS. The performance of C8, C18, and porous graphite columns were systematically evaluated and optimized for glycopeptide separation prior to mass spectrometry analysis by an LTQ Orbitrap XL ETD™.

仪器：

LTQ Orbitrap XL ETD hybrid mass spectrometer

结论：

• The Hypercarb porous graphite column demonstrated excellent separation for glycopeptide analysis, especially for short, hydrophilic peptides containing bi- or triantennary glycan chains. It allowed for their sensitive detection without any prior enrichment.

• Formation of metal adducts promoted evolution of higher-charge species, aiding ETD fragmentation of glycopeptides.

• ETD preserved labile glycans, facilitating the identification of both the peptide of interest and the site of modification.

• Combining peptide structural information obtained by ETD and the glycan composition information obtained by CID enabled confident identification and characterization of glycopeptides within a single LC-MS analysis using an LTQ Orbitrap XL ETD mass spectrometer.