Surface engineering on mesoporous silica chips for enriching low molecular weight phosphorylated proteins.
Surface engineering on mesoporous silica chips for enriching low molecular weight phosphorylated proteins.
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DOI：
10.1039/c0nr00720j
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发表时间：
2011-02
期刊：
影响因子：
6.7
通讯作者：
Ferrari M
中科院分区：
文献类型：
--
作者：
Hu Y;Peng Y;Lin K;Shen H;Brousseau LC 3rd;Sakamoto J;Sun T;Ferrari M
磷酸化肽和蛋白质在正常细胞活动中发挥重要作用，例如基因表达、有丝分裂、分化、增殖和凋亡，以及肿瘤的发生、进展和转移。然而，技术障碍阻碍了使用常见的分馏方法从复杂的生物液体（例如人血清）中捕获磷酸肽。在此，我们开发了一种双重策略材料，可增强对低分子量磷蛋白的捕获：具有精确设计孔径的介孔二氧化硅薄膜，可对分子尺寸进行空间选择，并结合针对磷蛋白的化学选择性表面修饰（即 Ga3+、Ti4+ 和 Zr4+）。这些材料具有高重现性（CV = 18%），并通过排除胰蛋白酶等降解酶来提高捕获蛋白质的稳定性。通过X射线衍射、透射电子显微镜、X射线光电子能谱、能量色散X射线光谱和椭圆光度法对复合介孔薄膜的化学和物理性质进行了表征。利用质谱和生物统计学分析，研究了介孔二氧化硅芯片上固定不同金属离子的富集效率。报道的新技术提供了一个能够有效分析血清蛋白质组的平台，用于生物标志物发现、法医采样和常规诊断应用。
Phosphorylated peptides and proteins play an important role in normal cellular activities, e.g., gene expression, mitosis, differentiation, proliferation, and apoptosis, as well as tumor initiation, progression and metastasis. However, technical hurdles hinder the use of common fractionation methods to capture phosphopeptides from complex biological fluids such as human sera. Herein, we present the development of a dual strategy material that offers enhanced capture of low molecular weight phosphoproteins: mesoporous silica thin films with precisely engineered pore sizes that sterically select for molecular size combined with chemically selective surface modifications (i.e. Ga3+, Ti4+ and Zr4+) that target phosphoroproteins. These materials provide high reproducibility (CV = 18%) and increase the stability of the captured proteins by excluding degrading enzymes, such as trypsin. The chemical and physical properties of the composite mesoporous thin films were characterized by X-ray diffraction, transmission electron microscopy, X-ray photoelectron spectroscopy, energy dispersive X-ray spectroscopy and ellipsometry. Using mass spectroscopy and biostatistics analysis, the enrichment efficiency of different metal ions immobilized on mesoporous silica chips was investigated. The novel technology reported provides a platform capable of efficiently profiling the serum proteome for biomarker discovery, forensic sampling, and routine diagnostic applications.
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影响因子：
46.9
作者：
Ficarro, SB;McCleland, ML;White, FM
通讯作者：
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影响因子：
3.4
作者：
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通讯作者：
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