用于血清中磷蛋白组分析的钛功能化交联壳聚糖聚合物的简便制备。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2024-10-15 DOI:10.1016/j.jchromb.2024.124347

Xiuqin Sheng, Yimin Guo, Chuan-Fan Ding, Yinghua Yan

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引用次数: 0

摘要

高效富集磷酸肽对蛋白质组学研究极为重要。在这项工作中，壳聚糖（CTs）、2,3-二羟基对苯二甲醛（2,3-DHA）和碳酰肼（CHZ）聚合生成聚合物（DHA-CTs-CHZ），然后将聚合物（DHA-CTs-CHZ）与大量钛离子耦合以富集磷酸肽。该材料具有高选择性（BSA 与 β-酪蛋白的比例为 5000:1M）、高灵敏度（0.001 fmol/μL）、高负载量（83.3 μg/mg）、高回收率（98.6 ± 1.2 %），并能有效地进行磷酸肽富集的尺寸排除。此外，还从正常人血清中成功捕获了与27种磷酸化蛋白相关的46种磷酸肽和31个磷酸化位点，而从阿尔茨海默病（AD）患者血清中成功捕获了与30种磷酸化蛋白相关的47种磷酸肽和35个磷酸化位点。

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Facile preparation of titanium functionalized cross-linked chitosan polymer for phosphoproteome analysis in serum

Efficient phosphopeptide enrichment is extremely important for proteomics research. In this work, chitosan (CTs), 2,3-dihydroxyterephthalaldehyde (2,3-DHA), and carbohydrazide (CHZ) are polymerized to generate the polymer (DHA-CTs-CHZ), and then the polymer (DHA-CTs-CHZ) is coupled with a significant number of titanium ions to enrich phosphopeptides. The material exhibits high selectivity (5000:1 M ratio of BSA to β-casein), sensitivity (0.001 fmol/μL), loading (83.3 μg/mg), recovery (98.6 ± 1.2 %), and effective size exclusion for phosphopeptide enrichment. In addition, 46 phosphopeptides and 31 phosphorylated sites associated with 27 phosphorylated proteins were successfully captured from the serum of normal subjects, while 47 phosphopeptides and 35 phosphorylated sites associated with 30 phosphorylated proteins were successfully captured from Alzheimer’s disease (AD) patients’ serum.

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.