Tsai Shuo-Wen, Loughran Michael, Suzuki Hiroaki, Karube Isao
Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
Electrophoresis. 2004 Feb;25(3):494-501. doi: 10.1002/elps.200305725.
Simultaneous electrophoresis of both native and Sodium dodecyl sulfate (SDS) proteins was observed on a single microchip within 20 min. The capillary array prevented lateral diffusion of SDS components and avoided cross contamination of native protein samples. The planar sputtered electrode format provided a more uniform distribution of separation voltage into each of the 36 parallel microchannel capillaries than platinum wire electrodes commonly used in conventional electrophoresis. The customized geometry of the stacking capillary machined into the cover plate of the microchip facilitated reproducible sample injection without the requirement for stacking gel. Polyimide served as a mask and facilitated insulation of the anode and cathode to prevent electrode lift off and deterioration during continuous electrophoresis, even at a constant current of 8 mA. Improved protein separation was observed during capillary electrophoresis at lower currents. Ferguson plot analysis confirmed the electrophoretic mobility of native globular proteins in accordance with their charge and size. Corresponding Ferguson plot analysis of SDS-associated proteins on the same chip confirmed separation of marker proteins according to their molecular weight.
在单个微芯片上20分钟内同时观察到天然蛋白质和十二烷基硫酸钠（SDS）蛋白质的电泳。毛细管阵列可防止SDS成分的横向扩散，并避免天然蛋白质样品的交叉污染。与传统电泳中常用的铂丝电极相比，平面溅射电极形式能为36个平行微通道毛细管中的每一个提供更均匀的分离电压分布。微芯片盖板上加工的堆积毛细管的定制几何形状有助于实现可重复的样品进样，而无需堆积凝胶。聚酰亚胺用作掩膜，有助于阳极和阴极的绝缘，以防止在连续电泳过程中电极脱落和劣化，即使在8 mA的恒定电流下也是如此。在较低电流下进行毛细管电泳时观察到蛋白质分离得到改善。弗格森图分析证实了天然球状蛋白质的电泳迁移率与其电荷和大小一致。在同一芯片上对SDS相关蛋白质进行相应的弗格森图分析，证实了标记蛋白质根据其分子量的分离情况。
