An aqueous two-phase system as a strategy for serum albumin depletion.
An aqueous two-phase system as a strategy for serum albumin depletion.
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DOI：
10.1016/j.talanta.2008.07.055
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发表时间：
2009-01
期刊：
影响因子：
6.1
通讯作者：
M. A. O. da Silva;M. Arruda
中科院分区：
文献类型：
--
作者：
M. A. O. da Silva;M. Arruda
基于浊点现象，提出了一种用于血浆白蛋白去除后的凝胶电泳（SDS-PAGE）分析的双水相体系。使用Triton®X-114和SDS的混合物评估白蛋白去除。首先优化了影响相分离和分配系数（K）的变量，如pH（从5.0到7.6）和盐组成，以在室温下进行相分离并促进白蛋白去除（15 min，约10 min）。95%的效率），以减少12%的α-螺旋结构，通过圆二色性分析证实的全球过程中，表面活性剂丰富的阶段。进行凝胶电泳分析，以比较粗品人血浆组成与接受浊点程序的人血浆以及接受使用基于色谱亲和柱的市售系统分离白蛋白的相同血浆。与粗血浆（10条蛋白质条带）相比，浊点提取程序可鉴定表面活性剂贫乏相中存在的18条蛋白质条带，从而获得更详细的电泳图谱。
An aqueous two-phase system, based on the cloud point phenomenon, was proposed for blood plasma albumin depletion prior to gel electrophoresis (SDS-PAGE) analysis. Albumin removal was evaluated using a mixture of Triton®X-114 and SDS. Variables affecting phase separation and the partition coefficient (K), such as pH (from 5.0 to 7.6) and salt composition, were first optimized to carry out phase separation at room temperature and to promote albumin removal (15min with ca. 95% efficiency) to the surfactant-rich phase, in a global process that results in reduction of 12% α-helix structures, confirmed through circular dichroism analysis. Gel electrophoresis analyses were carried out to compare crude human plasma composition with human plasma submitted to the cloud point procedure as well as the same plasma submitted to the separation of albumin using a commercially available system based on a chromatographic affinity column. The cloud point extraction procedure allowed identification of 18 protein bands present in the surfactant-poor phase, resulting in a more detailed electrophoretic profile, when compared with crude plasma (10 protein bands).