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用于微生物和生物聚合物毛细管等电聚焦的溶胶-凝胶柱技术,采用紫外或荧光检测。

Sol-gel column technology for capillary isoelectric focusing of microorganisms and biopolymers with UV or fluorometric detection.

作者信息

Horká Marie, Planeta Josef, Růzicka Filip, Slais Karel

机构信息

Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno.

出版信息

Electrophoresis. 2003 May;24(9):1383-90. doi: 10.1002/elps.200390177.

Abstract

The sol-gel surface modification is used for capillary isoelectric focusing of microorganisms and biopolymers. The coating strongly decreases the electroosmotic flow so that it enables the use of the short capillaries down to 100 mm in the separation length. The examples of capillary isoelectric focusing of the low-molecular-mass pI markers and mixed cultures of microbial populations of Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Enteroccocus faecalis with UV detection are shown. It is possible to quantify bacterial cells according to their peak areas; the minimum detectable number of microbial cells was 5 x 10(2)-1 x 10(3). The compatibility of sol-gel capillaries with sensitive fluorometric detection of fluorescent pI markers together with fluorescein labeled proteins is demonstrated.

摘要

溶胶-凝胶表面改性用于微生物和生物聚合物的毛细管等电聚焦。该涂层能显著降低电渗流,从而可使用分离长度低至100毫米的短毛细管。展示了低分子量等电点标记物以及大肠杆菌、白色念珠菌、表皮葡萄球菌和粪肠球菌微生物群体混合培养物的毛细管等电聚焦实例,并采用紫外检测。根据细菌细胞的峰面积可以对其进行定量;微生物细胞的最低可检测数量为5×10² - 1×10³ 。证明了溶胶-凝胶毛细管与荧光等电点标记物以及荧光素标记蛋白的灵敏荧光检测的兼容性。

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