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A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Published on: October 1, 2009
Bioinspired hydrophilic material with parallel ordered channels for simultaneously selective enrichment and rapid
Jie Li1, Junlong Wang2, Jiajun Cui1
1College of Chemistry and Materials Engineering, Zhejiang A & F University, Hangzhou, 311300, China.
Abstract:
Natural plants with fascinating structures give inspirations to fabricate advanced affinity chromatography materials for glycoproteome and phosphoproteome analyses. In this study, inspired by the long-range ordered structure and rapid water transportation capability of lotus stem, a biomimetic hydrophilic material (PE/HAVP-Ti4+) with parallel ordered channels is developed and applied to selectively enrich low-abundant glycopeptides and phosphopeptides in biological samples. The biomimetic PE/HAVP-Ti4+ has honeycomb porous structure, robust mechanical property, excellent hydrophilicity, and abundant affinity sites. It demonstrates the excellent selectivity (molar ratio human IgG: BSA = 1:1000, β-casein: BSA = 1:2500), high sensitivity (0.2 fmol μL-1 for glycopeptides, 0.01 fmol μL-1 for phosphopeptides), satisfactory recovery rate (90.7 % for glycopeptides, 91.1 % for phosphopeptides), and good recyclability (at least 10 cycles). Moreover, after enrichment treatment by PE/HAVP-Ti4+, a total of 411 N-glycopeptides assigned to 212 N-glycoproteins and 63 phosphopeptides associated with 41 phosphoproteins in human serum are identified by mass spectrometry. The PE/HAVP-Ti4+ has significant potential in applications of glycopeptide and phosphopeptide enrichment of real complex biological samples for the simultaneous study of glycoproteome and phosphoproteome.

