Comparison of alpha1-adrenergic receptor cell-membrane stationary phases prepared from expressed cell line and from rabbit hepatocytes |
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Authors: | Yu Wang Bingxiang Yuan Xiuling Deng Langchong He Sicen Wang Youyi Zhang Qide Han |
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Institution: | (1) Department of Pharmacology, Xi′an Jiaotong University Medical School, 710061 Xi′an, People’s Republic of China;(2) College of Pharmacy, Xi′an Jiaotong University, 710061 Xi′an, People’s Republic of China;(3) Institute of Vascular Medicine, Peking University Third Hospital, 100083 Beijing, People’s Republic of China |
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Abstract: | A G-protein-coupled receptor-cell-membrane stationary phase (CMSP) has been prepared by immobilizing cell membranes on the
surface of silica, as carrier. The resulting HEK293
α
1A
adrenoceptor cell-membrane stationary phase can be used for rapid on-line chromatographic determination of potential subtype-selective
α
1
-adrenoceptor ligand-binding affinities for α
1
-adrenoceptor subtypes. The objective of the research was to study whether cell lines stably overexpressing subtype receptors
could improve the sensitivity and specificity of cell-membrane chromatography (CMC) compared with use of homogenized tissue
and cells in primary culture. Effects of mobile-phase ionic strength, sample concentration, and the presence of competitive
agents on ligand-receptor interaction in CMSP were also evaluated. We found that cell lines stably overexpressing subtype
receptors led to improved sensitivity and specificity in CMC. The technique leads to useful procedures-cell-membrane stationary
phases may, for example, facilitate exploration of ligand-receptor interaction and determination of ligand-receptor binding
affinity in initial screening and separation of lead compounds or active components in Chinese traditional natural medicine
and herbs. This might eventually be an important contribution and an addition to our collection of techniques. |
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Keywords: | G-protein-coupled receptor Cell-membrane chromatography Cell-membrane stationary phase α 1A adrenergic receptors Immobilization |
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