Application of phenylboronic acid modified hydrogel affinity chips for high-throughput mass spectrometric analysis of glycated proteins.

Application of phenylboronic acid modified hydrogel affinity chips for high-throughput mass spectrometric analysis of glycated proteins.
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苯硼酸修饰水凝胶亲和芯片在糖化蛋白高通量质谱分析中的应用。

DOI:
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发表时间:
2007
影响因子:
2
通讯作者:
F. Schweigert
F. Schweigert
中科院分区:
化学3区
文献类型:
--
作者:
S. Gontarev;V. Shmanai;Simone K. Frey;M. Kvach;F. Schweigert

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翻译后修饰大大增加了人血浆蛋白质组的复杂性。除了生理变化,病理条件,如糖尿病是负责增加这种复杂性,通过产生晚期糖基化终产物(AGEs)。当搜索特定的生物标志物时,在分析之前降低这种复杂性是一个先决条件。为此,琼脂糖水凝胶用于在MALDI(基质辅助激光解吸/电离)靶的改性铝表面上产生高容量亲和层。3-氨基苯硼酸通过溴化氰活化固定作为配体的糖基化蛋白的亲和吸附,然后通过MALDI直接检测。实验结果表明,所制备的MALDI芯片蛋白容量高,分离效率高,非特异性蛋白结合率低。结果表明,苯硼酸改性的水凝胶是非常适合创建亲和表面的AGEs的高通量分析。
The complexity of the human plasma proteome is greatly increased by post-translational modifications. Besides physiological modifications, pathological conditions such as diabetes are responsible for adding to this complexity by producing advanced glycation endproducts (AGEs). When searching for specific biomarkers it is a prerequisite to reduce this complexity prior to analysis. To do this, agarose hydrogel was used to create a high-capacity affinity layer on the modified aluminum surface of MALDI (matrix-assisted laser desorption/ionization) targets. 3-Aminophenylboronic acid was immobilized via cyanogen bromide activation as a ligand for affinity sorption of glycated proteins, followed by their direct detection by MALDI. High protein capacity of prepared MALDI chips, efficient separation and low non-specific protein binding were demonstrated. The results show that phenylboronic acid modified hydrogels are very suitable for creating affinity surfaces for the high-throughput analysis of AGEs.
DOI: 10.1093/clinchem/45.5.593
发表时间: 1999-05
期刊: Clinical chemistry
影响因子: 9.3
作者:
D. Hage
通讯作者: D. Hage