Dual-functionalized two-dimensional metal–organic framework composite with highly hydrophilicity for effective enrichment of glycopeptides.

• Highly hydrophilic 2D MOF composite materials (Zr-Fc MOF@Au@GC) were successfully synthesized. • The 2-D morphology of Zr-Fc MOF@Au@GC was fully utilized, which significantly improved the enrichment efficiency. • The prepared material showed excellent sensitivity, selectivity, and binding capacity towards glycopeptides. • Zr-Fc MOF@Au@GC exhibited unprecedented enrichment performance in mode glycoproteins and human serum. Protein glycosylation research is currently focused on the development of various functionalized materials that can effectively enrich the levels of glycopeptides in samples. However, most of these materials possess limited glycopeptide-specific recognition sites because of large steric hindrance, unsuitable mass transfer kinetics, and relatively low surface areas. Herein, a highly hydrophilic two-dimensional (2-D) metal–organic framework (MOF) nanosheet modified with glutathione (GSH) and l -cysteine (l -Cys) (denoted as Zr-Fc MOF@Au@GC) has been synthesized for efficient glycopeptide enrichment. Using this composite material, 39 and 44 glycopeptides from horseradish peroxidase (HRP) and human serum immunoglobulin G (IgG) digests were detected, respectively, which represents a higher efficiency for glycopeptide enrichment from model glycoprotein digests than has been previously reported. The material Zr-Fc MOF@Au@GC exhibited ultra-high sensitivity (0.1 fmol/µL), excellent selectivity (weight ratio of HRP tryptic digest to bovine serum albumin (BSA) tryptic digest = 1:2000), good binding capacity (200 mg/g), satisfactory reusability, and long-term storage capacity. In addition, 655 glycopeptides corresponding to 366 glycoproteins were identified from human serum samples. To the best of our knowledge, this is the largest number of glycoproteins detected in human serum samples to date. These results indicated that Zr-Fc MOF@Au@GC has the potential to be used for the enrichment of glycopeptides in biological samples and the analysis of protein glycosylation. [ABSTRACT FROM AUTHOR]