One-step preparation of magnetic zwitterionic-hydrophilic dual functional nanospheres for in-depth glycopeptides analysis in Alzheimer's disease patients' serum.

A novel ultra-hydrophilic zwitterionic-HILIC (ZIC-HILIC) nanosphere (Fe3O4-CG) was synthesized via a one-step hydrothermal strategy, which significantly simplified the conventional multi-step procedures for the preparation of ZIC-HILIC materials. The dual-functional Fe3O4-CG nanosphere exhibited excellent selectivity (molar ratio BSA:HRP = 5000:1), low detection limit (0.05 fmol/µL), satisfactory reusability (at least 5 times) and recovery rate (93.7 ± 2.1%). The binding constant of Fe3O4-CG for HRP is 2.45 ± 0.32 × 10-6 M and the theoretical binding capacity is 330 mg g-1. In addition, the Fe3O4-CG microsphere showed excellent performance in the detection of glycopeptides from real biological samples. Furthermore, 131 glycopeptides related to 71 glycoproteins were selectively enriched from healthy human serum and 180 glycopeptides related to 82 glycoproteins were captured from Alzheimer's disease patients' serum analyzed by Nano-LC-MS/MS. Gene ontology analysis of the biological process and molecular function showed that 21 primitive glycoproteins in glycopeptides captured from Alzheimer's disease patients' serum were meaningfully involved in a variety of neurodegenerative disease-related events, including serine-type endopeptidase inhibitor activity, receptor binding, positive regulation of B cell activation, and platelet activation.