采用全柱荧光成像毛细管等速电泳研究蛋白质-蛋白质结合反应。
Study of protein-protein binding reaction by whole-column fluorescence-imaged CIEF.
作者信息
Wu Xing-Zheng, Asai Shinya, Yamaguchi Yoshie
机构信息
Department of Materials Science and Engineering, Graduate School of Engineering, University of Fukui, Bunkyo, Fukui-shi, Japan.
出版信息
Electrophoresis. 2009 May;30(9):1552-7. doi: 10.1002/elps.200800506.
Whole-column fluorescence-imaged CIEF was applied to study protein-protein binding reaction. A homemade whole-column fluorescence-imaged CIEF experimental setup was built, and its CIEF performance was evaluated with native fluorescent protein green fluorescence protein and fluorescently labeled proteins (bovine albumin, human albumin, and BSA). pIs, focusing time, detection limits, and linear quantitative range of the proteins were obtained. Furthermore, the method was used to study FITC-protein A-human IgG binding reaction. Experimental results showed that the apparent binding ratio of the FITC-protein A to human IgG was 1:2, and pI of the binding conjugates were about 6.3-6.5. No binding reaction was found between green fluorescence protein and the fluorescent-labeled proteins.
采用全柱荧光成像毛细管等速电泳（CIEF）研究蛋白质-蛋白质结合反应。搭建了自制的全柱荧光成像CIEF实验装置，并用天然荧光蛋白绿色荧光蛋白和荧光标记蛋白（牛血清白蛋白、人血清白蛋白和牛血清白蛋白）评估其CIEF性能。获得了蛋白质的等电点、聚焦时间、检测限和线性定量范围。此外，该方法还用于研究异硫氰酸荧光素-蛋白A-人免疫球蛋白结合反应。实验结果表明，异硫氰酸荧光素-蛋白A与人免疫球蛋白的表观结合比为1:2，结合缀合物的等电点约为6.3-6.5。未发现绿色荧光蛋白与荧光标记蛋白之间存在结合反应。
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