Li Jie, Wang Junlong, Cui Jiajun, Gao Fei, Guo Hongying, Yang Jiyou, Li Mengren, Ma Zhan, Wang Zijun, Zhang Liuping, Huan Weiwei
College of Chemistry and Materials Engineering, Zhejiang A & F University, Hangzhou, 311300, China.
College of Food and Health, Zhejiang A & F University, Hangzhou, 311300, China.
Talanta. 2025 Dec 1;295:128199. doi: 10.1016/j.talanta.2025.128199. Epub 2025 Apr 24.
Natural plants with fascinating structures give inspirations to fabricate advanced affinity chromatography materials for glycoproteome and phosphoproteome analyses. In this study, inspired by the long-range ordered structure and rapid water transportation capability of lotus stem, a biomimetic hydrophilic material (PE/HAVP-Ti) with parallel ordered channels is developed and applied to selectively enrich low-abundant glycopeptides and phosphopeptides in biological samples. The biomimetic PE/HAVP-Ti has honeycomb porous structure, robust mechanical property, excellent hydrophilicity, and abundant affinity sites. It demonstrates the excellent selectivity (molar ratio human IgG: BSA = 1:1000, β-casein: BSA = 1:2500), high sensitivity (0.2 fmol μL for glycopeptides, 0.01 fmol μL for phosphopeptides), satisfactory recovery rate (90.7 % for glycopeptides, 91.1 % for phosphopeptides), and good recyclability (at least 10 cycles). Moreover, after enrichment treatment by PE/HAVP-Ti, a total of 411 N-glycopeptides assigned to 212 N-glycoproteins and 63 phosphopeptides associated with 41 phosphoproteins in human serum are identified by mass spectrometry. The PE/HAVP-Ti has significant potential in applications of glycopeptide and phosphopeptide enrichment of real complex biological samples for the simultaneous study of glycoproteome and phosphoproteome.
具有迷人结构的天然植物为制备用于糖蛋白质组和磷酸化蛋白质组分析的先进亲和色谱材料提供了灵感。在本研究中,受莲藕茎的长程有序结构和快速输水能力的启发,开发了一种具有平行有序通道的仿生亲水材料(PE/HAVP-Ti),并将其应用于选择性富集生物样品中低丰度的糖肽和磷酸肽。仿生PE/HAVP-Ti具有蜂窝状多孔结构、强大的机械性能、优异的亲水性和丰富的亲和位点。它表现出优异的选择性(人IgG与BSA的摩尔比为1:1000,β-酪蛋白与BSA的摩尔比为1:2500)、高灵敏度(糖肽为0.2 fmol μL,磷酸肽为0.01 fmol μL)、令人满意的回收率(糖肽为90.7%,磷酸肽为91.1%)和良好的可重复使用性(至少10个循环)。此外,经PE/HAVP-Ti富集处理后,通过质谱鉴定出人血清中共有411个N-糖肽,归属212个N-糖蛋白,以及63个与41个磷酸化蛋白相关的磷酸肽。PE/HAVP-Ti在实际复杂生物样品的糖肽和磷酸肽富集应用中具有巨大潜力,可用于同时研究糖蛋白质组和磷酸化蛋白质组。