Capillary sieving electrophoresis-micellar electrokinetic chromatography fully automated two-dimensional capillary electrophoresis analysis of Deinococcus radiodurans protein homogenate
Capillary sieving electrophoresis-micellar electrokinetic chromatography fully automated two-dimensional capillary electrophoresis analysis of Deinococcus radiodurans protein homogenate
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DOI：
10.1002/elps.200405939
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发表时间：
2004-10-01
期刊：
影响因子：
2.9
通讯作者：
Dovichi, NJ
中科院分区：
文献类型：
--
作者：
Michels, DA;Hu, S;Dovichi, NJ
我们报道了耐辐射球菌蛋白匀浆的一维和二维（一维和二维）毛细管电泳分离。蛋白质用荧光试剂3-（2-呋喃基）喹啉-2-甲醛（FQ）标记，该试剂与2个赖氨酸残基反应并产生高荧光产物。检测是通过激光诱导荧光。一维毛细管筛分电泳（CSE）生产超过15万个板，胶束电动毛细管色谱（MEKC）生产超过90万个板，用于检测D. radiodurans蛋白匀浆中的成分。在二维分离中，蛋白质首先通过CSE分离。分数重复转移到第二毛细管进一步分离基于MEKC。二维分离具有类似于550个点的容量。超过150种成分从匀浆中部分分离出来。在第一个维度上，由于长分离周期内蛋白质的扩散，分辨率受到限制；在第二个维度上，由于长组分转移时间和短分离周期的结合，分辨率受到限制。
We report the one- and two-dimensional (1-D and 2-D) capillary electrophoresis separation of Deinococcus radiodurans protein homogenate. Proteins are labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ), which reacts with 2 lysine residues and creates a highly fluorescent product. Detection is by laser-induced fluorescence. 1-D capillary sieving electrophoresis (CSE) produces over 150 000 plates and micellar electrokinetic capillary chromatography (MEKC) produces over 900000 plates for components in a D. radiodurans protein homogenate. In a 2-D separation, proteins are first separated by CSE. Fractions are repetitively transferred to a second capillary for further separation based on MEKC. The 2-D separation has a similar to550 spot capacity. Over 150 components are partially resolved from the homogenate. Resolution is limited in the first dimension by diffusion of proteins during the long separation period and in the second dimension by the combination of a long fraction-transfer time and short separation period.