用聚乙烯亚胺和植酸官能化的亲水性 Ti4+-immobilized 树枝状聚(甲基丙烯酸缩水甘油酯)微粒一步富集和逐步洗脱糖蛋白和磷蛋白。

IF 2.8 3区 工程技术 Q2 CHEMISTRY, ANALYTICAL Journal of separation science Pub Date : 2024-07-01 DOI:10.1002/jssc.202400154
Hao Wang, Simin Li, Xinyan Wang, Xinyue Li, Minzhi Jiang, Shiying Chen, Zaiqi Hu, Haitian Li, Yuzhen Xu, Lei Jin
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引用次数: 0

摘要

糖基化和磷酸化是最重要的翻译后修饰,对它们的分析在很大程度上依赖于富集技术。在这项工作中,我们开发了一种简便的方法,可一步同时富集和逐步洗脱糖蛋白和磷酸化蛋白。该方法的核心是应用新型钛(IV)离子固定化聚(甲基丙烯酸缩水甘油酯)微粒,该微粒具有树枝状聚乙烯亚胺和植酸功能。由于微颗粒表面含有丰富的钛离子和羟基,因此具有双重富集能力。它们显示出快速的吸附平衡(30 分钟内)以及对 β-酪蛋白(1107.7 毫克/克)和辣根过氧化物酶(438.6 毫克/克)的超强吸附能力,超过了牛血清白蛋白(91.7 毫克/克)。此外,还进行了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳来验证富集能力。对各种生物样本(包括标准蛋白质混合物、脱脂牛奶和人血清)的实验结果表明,这些微粒具有从生物样本中富集低丰度糖蛋白和磷蛋白的显著能力。
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One-step enrichment and stepwise elution of glycoproteins and phosphoproteins by hydrophilic Ti4+-immobilized dendrimer poly(glycidyl methacrylate) microparticles functionalized with polyethylenimine and phytic acid

Glycosylation and phosphorylation rank as paramount post-translational modifications, and their analysis heavily relies on enrichment techniques. In this work, a facile approach was developed for the one-step simultaneous enrichment and stepwise elution of glycoproteins and phosphoproteins. The core of this approach was the application of the novel titanium (IV) ion immobilized poly(glycidyl methacrylate) microparticles functionalized with dendrimer polyethylenimine and phytic acid. The microparticles possessed dual enrichment capabilities due to their abundant titanium ions and hydroxyl groups on the surface. They demonstrate rapid adsorption equilibrium (within 30 min) and exceptional adsorption capacity for β-casein (1107.7 mg/g) and horseradish peroxidase (438.6 mg/g), surpassing that of bovine serum albumin (91.7 mg/g). Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis was conducted to validate the enrichment capability. Experimental results across various biological samples, including standard protein mixtures, non-fat milk, and human serum, demonstrated the remarkable ability of these microparticles to enrich low-abundance glycoproteins and phosphoproteins from biological samples.

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来源期刊
Journal of separation science
Journal of separation science 化学-分析化学
CiteScore
6.30
自引率
16.10%
发文量
408
审稿时长
1.8 months
期刊介绍: The Journal of Separation Science (JSS) is the most comprehensive source in separation science, since it covers all areas of chromatographic and electrophoretic separation methods in theory and practice, both in the analytical and in the preparative mode, solid phase extraction, sample preparation, and related techniques. Manuscripts on methodological or instrumental developments, including detection aspects, in particular mass spectrometry, as well as on innovative applications will also be published. Manuscripts on hyphenation, automation, and miniaturization are particularly welcome. Pre- and post-separation facets of a total analysis may be covered as well as the underlying logic of the development or application of a method.
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