A DNA tetrahedron-inspired magnetic hybrid with regular distribution of trypsin for ultra-fast digestion of proteins and glycoproteomics analysis

Abstract

In large-scale glycoproteomics studies, reducing the experimental duration and enhancing digestion efficiency are crucial. In this work, a magnetic nanomaterial featuring a surface rich in hydrophilic groups and uniformly distributed trypsin (Fe₃O₄@DOPA/PEI@Au@DNA TET-trypsin) was developed for efficient digestion of proteins and specific enrichment of glycopeptides. The material surface exhibits a regular distribution of trypsin coupled with abundant hydrophilic groups. Thus, the material can demonstrate excellent performance in experimental studies related to glycoproteomics. Fe₃O₄@DOPA/PEI@Au@DNA TET-trypsin showed an excellent limit of detection (0.001 amol·μL⁻¹) and high selectivity for glycopeptides (BSA : HRP = 5000 : 1) in a short reaction time, and it showed good performance in the processing of practical samples. Gene Ontology (GO) analysis suggests that complement activation, extracellular space, antioxidant activity and complement binding may be associated with tuberculosis. These analytical results provide new insights for determining and clarifying the pathogenesis and pathology related to tuberculosis.