Light-/pH-Regulated Spiropyran Smart-Responsive Hydrophilic Separation Platform for the Identification of Serum Glycopeptides from Hepatocellular Carcinoma Patients

Smart-responsive materials have attracted much attention in the enrichment of post-translational modifications of proteins. In this work, for the first time, we developed a smart enrichment strategy (MNPs-l-DOPA/PEI-SP) based on the change in hydrophilic properties of spiropyran under the regulation of light and pH to realize the controllable enrichment and release of intact glycopeptides. The enrichment mechanism and possible binding mechanism were verified by theoretical calculations. The smart enrichment platform based on MNPs-l-DOPA/PEI-SP was used to screen glycoprotein biomarkers for hepatocellular carcinoma (HCC) to evaluate its cancer diagnostic and monitoring performance. A total of 3,864 intact N-glycopeptides containing 166 N-glycoproteins were successfully identified in serum samples of early-stage HCC patients, while 3,266 intact N-glycopeptides containing 193 glycoproteins were identified in normal control (NC) serum samples. This work not only provides new ideas for the efficient enrichment of intact glycopeptides with smart-responsive material, but also broadens the research possibilities for biomarker discovery in HCC serum liquid biopsies.