Bioinspired Hydrophilic Material with Parallel Ordered Channels for Simultaneously Selective Enrichment and Rapid Separation of Glycopeptides and Phosphopeptides

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 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 using PE/HAVP-Ti4+ treatment, 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 the application of glycopeptide and phosphopeptide enrichment in real complex biological samples for the simultaneous study of glycoproteome and phosphoproteome researches.

Keywords: Glycosylation, Phosphorylation, Biomimetic material, Ordered structure, Enrichment, Mass Spectrometry