Alzheimer's disease (AD) is a common neurodegenerative disease. Abnormal glycosylation and phosphorylation modification in AD may be closely related to its pathology. It is of substantial practical significance to simultaneously investigate the roles of phosphorylation and glycosylation in AD. In this work, a bifunctional super-hydrophilic mesoporous nanocomposite (denoted mTiO2@AuCG) was prepared, which combined hydrophilic interaction chromatography (HILIC) and metal oxide affinity chromatography (MOAC) enrichment strategies to enrich phosphopeptides and glycopeptides, respectively or simultaneously. The mTiO2@AuCG exhibited excellent performance on the high-efficiency enrichment of glycopeptides (selectivity, 5000:1 molar ratios of BSA/HRP; sensitivity, 0.1 fmol HRP; satisfactory recovery rate; loading capacity, 200 mg/g) and phosphopeptides (selectivity, 1000:1 molar ratios of BSA/β-casein; sensitivity, 0.2 fmol β-casein; satisfactory recovery rate; loading capacity, 200 mg/g). Using these advantages, after single-step enrichment of mTiO2@AuCG, a total of 209 glycopeptides related to 93 glycoproteins, and 17 phosphopeptides related to 13 phosphoproteins were detected from normal human serum. By contrast, 167 glycopeptides related to 88 glycoproteins, and 14 phosphopeptides related to 12 phosphoproteins were found in AD serum.
Keywords: Alzheimer's disease; Glycosylation and phosphorylation analysis; Mesoporous nanocomposite; Nano-LC-MS/MS; mTiO(2)@AuCG.
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