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使用过表达内皮素受体的固定化全细胞进行毛细管电泳以筛选特异性配体。

Capillary electrophoresis using immobilized whole cells with overexpressed endothelin receptor for specific ligand screening.

作者信息

Yu Hui-Ming, Tseng Min-Jen, Fang Jim-Min, Phutrakul Suree, Chen Shui-Tein

机构信息

Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.

出版信息

Electrophoresis. 2004 Apr;25(7-8):1034-41. doi: 10.1002/elps.200305804.

Abstract

A new capillary electrophoresis method using immobilized cells as the stationary phase has been developed. The power of this method is demonstrated by the separation and identification of endothelin antagonists on a capillary column coated by the transfected Chinese hamster ovary (CHO) cells with overexpressing endothelin receptors. The screening results are validated by functional assays suppressing the increase of intracellular calcium concentration induced by endothelin-1. Instead of making efforts in isolating protein receptors, the easily prepared whole-cell capillary column provides a superior tool on the basis of ligand/receptor affinity for a rapid screening of potent drug candidates from compound libraries.

摘要

一种以固定化细胞作为固定相的新型毛细管电泳方法已被开发出来。通过在涂有过表达内皮素受体的转染中国仓鼠卵巢(CHO)细胞的毛细管柱上分离和鉴定内皮素拮抗剂,证明了该方法的有效性。筛选结果通过抑制内皮素-1诱导的细胞内钙浓度升高的功能测定得到验证。无需费力分离蛋白质受体,易于制备的全细胞毛细管柱基于配体/受体亲和力提供了一种卓越的工具,可用于从化合物库中快速筛选有效的候选药物。

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