Ultrasensitive Characterization of Site-Specific Glycosylation of Affinity-Purified Haptoglobin from Lung Cancer Patient Plasma Using 10 μm i.d. Porous Layer Open Tubular Liquid Chromatography−Linear Ion Trap Collision-Induced Dissociation/Electron Transfer Dissociation Mass Spectrometry
| Autor: | Tomas Rejtar, Dongdong Wang, Barry L. Karger, Marina Hincapie |
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| Rok vydání: | 2011 |
| Předmět: | |
| Zdroj: | Analytical Chemistry. 83:2029-2037 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac102825g |
| Popis: | Site-specific analysis of protein glycosylation is important for biochemical and clinical research efforts. Glycopeptide analysis using liquid chromatography−collision-induced dissociation/electron transfer dissociation mass spectrometry (LC−CID/ETD-MS) allows simultaneous characterization of the glycan structure and attached peptide site. However, due to the low ionization efficiency of glycopeptides during electrospray ionization, 200−500 fmol of sample per injection is needed for a single LC−MS run, which makes it challenging for the analysis of limited amounts of glycoprotein purified from biological matrixes. To improve the sensitivity of LC−MS analysis for glycopeptides, an ultranarrow porous layer open tubular (PLOT) LC column (2.5 m × 10 μm i.d.) was coupled to a linear ion trap (LTQ) collision-induced dissociation/electron transfer dissociation mass spectrometer to provide sensitive analysis of N-linked protein glycosylation heterogeneity. The potential of the developed method is demonstrated by ... |
| Databáze: | OpenAIRE |
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