Silvertand L H H, Machtejevas E, Hendriks R, Unger K K, van Bennekom W P, de Jong G J
Department of Biomedical Analysis, Faculty of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Jul 24;839(1-2):68-73. doi: 10.1016/j.jchromb.2006.03.036. Epub 2006 Apr 4.
This paper describes the on-line sample pretreatment and analysis of proteins and peptides with a poly(methylmethacrylate) (PMMA) microfluidic device (IonChip). This chip consists of two hyphenated electrophoresis channels with integrated conductivity detectors. The first channel can be used for sample preconcentration and sample clean-up, while in the second channel the selected compounds are separated. Isotachophoresis (ITP) combined with zone electrophoresis (CZE) was used to preconcentrate a myoglobin sample by a factor of about 65 before injection into the second dimension and to desalt a mixture of six proteins with 100 mM NaCl. However, ITP-CZE could not be used for the removal of two proteins from a protein/peptide sample since the protein zone in the ITP step was too small to remove certain compounds. Therefore, we used CZE-CZE for the removal of proteins from a protein/peptide mixture, thereby injecting only the peptides into the second CZE separation channel.
本文描述了使用聚甲基丙烯酸甲酯（PMMA）微流控装置（离子芯片）对蛋白质和肽进行在线样品预处理及分析。该芯片由两个带有集成电导检测器的联用电泳通道组成。第一个通道可用于样品预浓缩和样品净化，而在第二个通道中对选定的化合物进行分离。等速电泳（ITP）与区带电泳（CZE）相结合，在将肌红蛋白样品注入第二维之前将其预浓缩约65倍，并对含有100 mM NaCl的六种蛋白质混合物进行脱盐。然而，ITP-CZE不能用于从蛋白质/肽样品中去除两种蛋白质，因为ITP步骤中的蛋白质区带太小，无法去除某些化合物。因此，我们使用CZE-CZE从蛋白质/肽混合物中去除蛋白质，从而仅将肽注入第二个CZE分离通道。
