Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
J Pharm Biomed Anal. 2010 Dec 15;53(5):1272-7. doi: 10.1016/j.jpba.2010.07.008. Epub 2010 Aug 1.
A simple method for a chemical surface modification of poly(methyl methacrylate) (PMMA) microchips with amino-poly(ethyleneglycol) (PEG-NH(2)) by nucleophilic addition-elimination reaction was developed to improve the separation efficiency and analytical reproducibility in a microchip electrophoresis (MCE) analysis of biomolecules such as proteins and enantiomers. In our procedure, the PEG chains were robustly immobilized only by introducing an aqueous solution of PEG-NH(2) into the PMMA microchannel. The electroosmotic mobilities on the modified chips remained almost constant during 35 days with 37 runs without any recoating. The PEG-NH(2) modified chip provided a fast, reproducible, efficient MCE separation of proteins with a wide variety of isoelectric points within 15s. Furthermore, the application of the modified chip to affinity electrophoresis using bovine serum albumin gave a good chiral separation of amino acids.
一种通过亲核加成-消除反应将氨基聚(乙二醇)(PEG-NH2)化学表面修饰到聚甲基丙烯酸甲酯(PMMA)微芯片上的简单方法,旨在提高生物分子(如蛋白质和对映体)在微芯片电泳(MCE)分析中的分离效率和分析重现性。在我们的方法中,仅通过将 PEG-NH2 的水溶液引入 PMMA 微通道,就可以牢固地固定 PEG 链。在 37 次运行且无需重新涂覆的情况下,经过 35 天,修饰后的芯片上的电渗流迁移率几乎保持不变。PEG-NH2 修饰的芯片可在 15s 内快速、重现、有效地分离具有各种等电点的蛋白质。此外,将修饰后的芯片应用于使用牛血清白蛋白的亲和电泳,可很好地分离氨基酸对映体。