ELECTROPHORETIC SEPARATIONS OF PROTEINS IN CAPILLARIES WITH HYDROLYTICALLY STABLE SURFACE-STRUCTURES
ELECTROPHORETIC SEPARATIONS OF PROTEINS IN CAPILLARIES WITH HYDROLYTICALLY STABLE SURFACE-STRUCTURES
复制标题
DOI：
10.1021/ac00221a013
复制
发表时间：
1990-11-15
影响因子：
7.4
通讯作者：
NOVOTNY, M
中科院分区：
文献类型：
--
作者：
COBB, KA;DOLNIK, V;NOVOTNY, M
描述了一种获得用于毛细管电泳（CE）的高度稳定的涂覆毛细管的方法。表面氯化熔融二氧化硅毛细管与格氏试剂乙烯基溴化镁反应，随后乙烯基与丙烯酰胺反应，通过水解稳定的Si-C键形成聚丙烯酰胺固定层。这种方法是Hjerten先前描述的毛细管涂层过程的延伸，不同的是聚丙烯酰胺层与毛细管壁结合的方式。以本文描述的方式处理的毛细管可以在2-10.5的pH范围内使用，而不会引起涂层的明显分解。与未涂覆的毛细血管相比，由于蛋白质对毛细血管壁的吸附减少，使用这种涂覆的毛细血管的蛋白质分离得到了改善，尽管在某种程度上仍然存在相互作用，如无法获得理论预测的高板计数所证明。与未涂覆的毛细血管相比，涂覆后的毛细血管中几乎消除了电渗透流动，从而提高了蛋白质迁移时间的可重复性。此外，对模型蛋白的峰偏度进行了评估，并指出了涂层毛细血管的改进。结果提出了模型蛋白混合物的分离，比较了乙烯基结合的聚丙烯酰胺涂层毛细血管和未涂层毛细血管在高和低pH值极端下的性能。
A procedure for obtaining highly stable coated capillaries for use in capillary electrophoresis (CE) is described. Reaction of surface-chlorinated fused silica capillaries with the Grignard reagent, vinyl magnesium bromide, followed by reaction of the vinyl group with acrylamide, results in an immobolized layer of polyarylamide attached through hydrolytically stable Si-C bonds. This method is an extension of the capillary coating procedure described previously by Hjerten, differing in the means by which the polyacrylamide layer is bonded to the capillary walls. Capillaries treated in the manner described here can be used over a pH range of 2-10.5, without noticeable decomposition of the coating. In comparison to uncoated capillaries, separations of proteins using such coated capillaries are improved due to a reduction in protein adsorption to the capillary walls, although interaction is still present to some degree as evidenced by an inability to obtain plate counts as high as those predicted by theory. Electroosmotic flow is virtually eliminated in the coated capillaries, resulting in improved reproducibilities of protein migration times in comparison to uncoated capillaries. Additionally, peak skew is evaluated for model proteins and improvements are noted for the coated capillaries. Results are presented for separations of model protein mixtures, comparing the performance of the vinyl-bound polyacrylamide coated capillaries and uncoated capillaries at both high and low pH extremes.