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

IF 3.6 3区化学 Q2 CHEMISTRY, ANALYTICAL

Analyst Pub Date : 2025-05-28 DOI:10.1039/d5an00357a

Jiahui Lai, Danni Wang, Xiuqin Sheng, Chuanfan Ding, Yinghua Yan

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Abstract

In large-scale glycoproteomics studies, reducing the experimental duration and enhancing the digestion efficiency are crucial for the experiments. 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.

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DNA四面体激发磁杂交与规则分布的胰蛋白酶超快速消化蛋白质和糖蛋白组学分析

在大规模糖蛋白组学研究中，缩短实验时间和提高消化效率是实验的关键。在这项工作中，开发了一种磁性纳米材料，其表面富含亲水性基团和均匀分布的胰蛋白酶（Fe3O4@DOPA/PEI@Au@DNA TET-Trypsin），用于有效消化蛋白质和特异性富集糖肽。材料表面有规则分布的胰蛋白酶和丰富的亲水性基团。因此，该材料在糖蛋白组学相关的实验研究中表现出优异的性能。Fe3O4@DOPA/PEI@Au@DNA TET-Trypsin在较短的反应时间内对糖肽类（BSA:HRP=5000:1）具有良好的检出限（0.001 amol·μL-1）和选择性，在实际样品的处理中表现出良好的性能。基因本体（GO）分析表明，补体活化、细胞外空间、抗氧化活性和补体结合可能与结核病有关。这些分析结果为确定和阐明结核病的发病机制和病理提供了新的见解。

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