8-Gluconolactone was covalently coupled to aminopropyl derivatized capillary, which created hydrophilic brushes on the inner wall of the capillary. The coated capillary was shown to generate a stable electroosmotic flow （EOF） in the investigated pH range of 2.0-9.0 and to suppress effectively the adsorption of proteins. And it enabled separation of some biopolymer mixtures including basic proteins, DNA and tryptic digested bovine serum albumin （BSA） within 15 rain with efficiencies up to 450,000 plates/m. The intra- and inter-day reproducibility of the coating referring to the retention times of proteins were satisfactory with mean relative standard deviations （R.S.D.） of 0.8 and 1.7%, respectively. 2009 Yin Mao Wei. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All fights reserved.
Lian Guo ShanXue YuYin Mao WeiXiao Hui ZhengJian Bin Zheng
β2-肾上腺素能受体是细胞表面受体的一种,它能通过偶联异源三聚体G蛋白将信号转导引入到细胞内部。本实验在成功克隆、表达β2-肾上腺素能受体的基础上,建立了一种两步柱色谱分离纯化目的蛋白质的方法。首先利用Ni2+螯合的高分辨纯化的预带电荷介质Sepharose High Performance与含有六聚组氨酸标签的蛋白质特异结合的性质,对目的蛋白质进行初步分离,接着运用快流速Q琼脂糖凝胶(Quaternary Sepharose FastFlow)对其进行进一步的分离纯化。采用该方法得到的β2-肾上腺素能受体蛋白质经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和高效凝胶排阻色谱检测其纯度约为95%。结果表明该方法可以对重组猪β2-肾上腺素能受体进行有效的分离纯化。
