葡萄糖-6-磷酸功能化亲水磁探针:有效分离富集糖肽/磷酸肽的双用途亲和材料  

Glucose-6-phosphate Functionalized Hydrophilic Magnetic Probe:a Dual-purpose Affinity Material for Effective Separation and Enrichment of Glycopeptides/Phosphopeptides

作　　者：王柏纯 袁雨欣 闫迎华 丁传凡 唐科奇 WANG Baichun;YUAN Yuxin;YAN Yinghua;DING Chuanfan;TANG Keqi(Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis of Zhejiang Province,Institute of Mass Spectrometry,School of Material Science and Chemical Engineering,Ningbo University,Ningbo 315211,China)

机构地区：[1]浙江省先进质谱技术与分子检测重点实验室,宁波大学材料科学与化学工程学院质谱技术与应用研究院,宁波315211

出　　处：《高等学校化学学报》2021年第10期3062-3070,共9页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:21927805);国家重点研发计划项目(批准号:2016YFF0102900);宁波市自然科学基金(批准号:2018A610180);宁波大学王宽诚基金资助.

摘　　要：在Fe_(3)O_(4)磁芯上通过逐层修饰构建了葡萄糖-6-磷酸(G6P)功能化亲水磁探针Fe_(3)O_(4)@PDA@TiO_(2)@G6P.聚多巴胺(PDA)可以作为偶联连接剂进一步接枝二氧化钛(TiO_(2));接枝的TiO_(2)除作为G6P的锚定位点外,还可通过金属氧化物亲和层析技术有效富集磷酸肽;G6P的官能化赋予了纳米球高亲水性的表面,并利用亲水作用液相色谱法实现了糖肽的捕捉.实验结果表明,探针对糖肽具有低的检出限(0.1 fmol/μL)、高的选择性[m(HRP):m(BSA)=1:1000]、良好的重复性(10次循环)和高的负载量(300 mg/g);对磷酸肽具有低的检出限(0.02 fmol/μL)和高的选择性[n(β-casein):n(BSA)=1:1000).此外,这种双用途亲和材料具有同时富集糖肽和磷酸肽的能力,可从人唾液中鉴定出34个糖肽和36个磷酸肽,表明其在多种翻译后修饰的蛋白质组学分析中具有巨大的应用前景.Glucose-6-phosphate(G6P)functionalized hydrophilic magnetic probe Fe_(3)O_(4)@PDA@TiO_(2)@G6P was constructed by the layer-by-layer modification on the surface of the magnetic core Fe_(3)O_(4).Polydopamine(PDA)not only enhanced the hydrophilicity of the probe,but also acted as the coupling linker for the further grafting of titanium dioxide(TiO_(2)).In addition to serving as the anchor point of G6P,the grafted TiO_(2) could al⁃so effectively enrich phosphopeptides through the metal oxide affinity chromatography technology.The func⁃tionalization of G6P gave the nanospheres a highly hydrophilic surface and used the hydrophilic interaction liquid chromatography technology to achieve the goal of capturing glycopeptides.The results showed that this dual-purpose affinity probe had a low detection limit(0.1 fmol/μL),a high selectivity[m(HRP):m(BSA)=1:1000],a good repeatability(ten cycles)and a high loading(300 mg/g)for glycopeptides.Besides,as for phos⁃phopeptides,the obtained G6P functionalized hydrophilic magnetic probe exhibited a low detection limit(0.02 fmol/μL)and a high selectivity[n(β-casein):n(BSA)=1:1000].At the same time,this dual-purpose affinity material had the ability to simultaneously enrich glycopeptides and phosphopeptides,and 34 glycopeptides and 36 phosphopeptides were identified from human saliva,indicating that it will show great application pros⁃pects in proteomics analysis of a variety of post-translational modifications.

关 键 词：双用途亲水磁探针 葡萄糖-6-磷酸 糖肽/磷酸肽 质谱检测 

分 类 号：O657[理学—分析化学]