Free-flow electrophoresis in a microfabricated chamber with a micromodule fraction separator - Continuous separation of proteins
Free-flow electrophoresis in a microfabricated chamber with a micromodule fraction separator - Continuous separation of proteins
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DOI：
10.1016/s0021-9673(02)01964-7
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发表时间：
2003-03-21
影响因子：
4.1
通讯作者：
Shinohara, E
中科院分区：
文献类型：
--
作者：
Kobayashi, H;Shimamura, K;Shinohara, E
蛋白质的连续自由流动电泳法是在微型自由流动电泳仪(MFFE)中进行的，分离物有30微米厚的狭缝。新研制的微模分数分离器(MFS)安装在mFFE分离室的下游端部。通过使用MFS，将来自分离室的电解液引入到蠕动泵中，而不干扰分离室底部的电解液流动。蛋白质混合物样品的分离是通过分离室的羟丙甲基纤维素前处理涂层来实现的。样品室的前处理有效地抑制了电渗流。所有分离样品经连续电泳法收集后，用反相高效液相色谱法进行分析。从高效液相分析的结果来看，其他端口的细胞色素c组分在相邻端口都没有发现串扰现象。肌红蛋白也有类似的结果。这意味着这些蛋白质被连续的mFFE完全分离，并且MFS在连续电泳过程中有效地发挥作用。(C)2002 Elsevier Science B.V.保留所有权利。
Continuous free flow electrophoresis of proteins was carried out in a microfabricated free flow electrophoresis (mFFE) module with the 30-mum thick slit of the separation. The newly developed micromodule fraction separator (MFS) was attached to the down-stream end site of the separation chamber of mFFE. By using the MFS, electrolyte solution from the separation chamber was introduced to the peristaltic pump without disturbing the electrolyte solution flow at the bottom side of the chamber. The separation of protein mixture samples was achieved by a hydroxypropylmethylcellulose pretreatment coating of the separation chamber. The pretreatment of the sample chamber effectively suppressed electroosmotic flow. All fractionated samples were collected using the MFS after continuous elecrophoresis and analyzed by reversed-phase HPLC. From the results of HPLC analyses none of the cytochrome c fractions at the other ports revealed cross talk phenomena at adjacent ports. A similar result occurred for the myoglobin. This means that these proteins were completely separated from each other by continuous mFFE, and the MFS functioned efficiently during continuous electrophoresis. (C) 2002 Elsevier Science B.V. All rights reserved.