Monolithic porous polymer layer for the separation of peptides and proteins using thin-layer chromatography coupled with MALDI-TOF-MS
Monolithic porous polymer layer for the separation of peptides and proteins using thin-layer chromatography coupled with MALDI-TOF-MS
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DOI：
10.1021/ac061527i
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发表时间：
2007-01-15
影响因子：
7.4
通讯作者：
Svec, Frantisek
中科院分区：
文献类型：
--
作者：
Bakry, Rania;Bonn, Gunther K.;Svec, Frantisek
制备了具有多孔聚合物整体层的薄层色谱（TLC）板，并用于分离小分子、肽和蛋白质。50-200亩。使用UV引发的聚合原位制备薄的聚（甲基丙烯酸丁酯-共-二甲基丙烯酸乙烯酯）层。精确控制反应条件可以制备具有明确多孔结构的整体层，从而决定色谱性能。与使用基于二氧化硅的预涂层的常规TLC和高性能TLC相比，预期小的层厚度和不存在任何粘合剂将改善基于聚合物的整体薄层色谱板的保留特性和分离效率。首先使用典型的UV成像检测分离的化合物的斑点。由于单片薄层也可以直接在不锈钢MALDI载体板上制备，TLC形式的分离可以与MALDI-TOF-MS结合。常规MALDI基质的应用促进了肽和蛋白质的解吸和电离，用于分离化合物的分子量测定。
Plates for thin-layer chromatography (TLC) with an attached layer of porous polymer monolith have been prepared and used for the separation of small molecules, peptides, and proteins. The 50-200-mu m. thin poly(butyl methacrylate-co-ethylene dimethacrylate) layers were prepared in situ using UV-initiated polymerization. Precise control of the reaction conditions enables the preparation of monolithic layers with a well-defined porous structure that determines the chromatographic performance. Compared to conventional TLC and high-performance TLC using precoated layers based on silica, the small layer thickness and absence of any binder is expected to improve both retention characteristics and separation efficiency of the polymer-based monolithic thin-layer chromatographic plates. Spots of the separated compounds were first detected using typical UV imaging. Since the monolithic thin layers can be also prepared directly on the stainless steel MALDI carrier plate, the separation in TLC format can be coupled with MALDI-TOF-MS. Application of a conventional MALDI matrix facilitated desorption and ionization of peptides and proteins for molecular weight determination of the separated compounds.