Cong Yongzheng, Zhang Lihua, Tao Dingyin, Liang Yu, Zhang Weibing, Zhang Yukui
National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, China.
J Sep Sci. 2008 Feb;31(3):588-94. doi: 10.1002/jssc.200700444.
A two-dimensional capillary electrophoresis platform, combining isoelectric focusing (IEF) and capillary zone electrophoresis (CZE), was established on a microchip with the channel width and depth as 100 mum and 40 mum, respectively. With polyacrylamide as permanent coating, EOF in the microchannel, which could impair the separation, was decreased to 3.4x10(-9)m(2).V(-1).s(-1), about 1/10 of that obtained in the uncoated set-up. During the separation, peptides were first focused by IEF in the first dimensional channel, and then directly driven into the perpendicular channel by controlling the applied voltages, and separated by CZE. Effects of various experimental parameters, including the electric field strength, channel length, and injection frequency from the first to the second dimensional separation channel, were studied. Under optimized condition, the digests of BSA and proteins extracted from E. coli were separated, and a peak capacity of 540 was obtained, which was far greater than that obtained by each single dimensional separation. All these results showed the promise of multidimensional separation on a microchip for the high-throughput and high-resolution analysis of complex samples.
在通道宽度和深度分别为100μm和40μm的微芯片上建立了一种二维毛细管电泳平台，该平台结合了等电聚焦（IEF）和毛细管区带电泳（CZE）。以聚丙烯酰胺作为永久性涂层，微通道中可能会影响分离的电渗流降低至3.4×10⁻⁹m²·V⁻¹·s⁻¹，约为未涂层装置中所得值的1/10。在分离过程中，肽首先在第一维通道中通过IEF聚焦，然后通过控制施加电压直接驱动到垂直通道中，并通过CZE进行分离。研究了各种实验参数的影响，包括电场强度、通道长度以及从第一维到第二维分离通道的进样频率。在优化条件下，对牛血清白蛋白（BSA）的酶解产物和从大肠杆菌中提取的蛋白质进行了分离，获得了540的峰容量，这远大于通过单维分离获得的峰容量。所有这些结果表明了微芯片上多维分离在复杂样品高通量和高分辨率分析方面的前景。
