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学科主题: 临床医学
题名:
The preparation of HEK293 alpha(1A) or HEK293 alpha(1B) cell membrane stationary phase and the chromatographic affinity study of ligands of alpha(1) adrenoceptor
作者: Wang, Y; Yuan, BX; Deng, XL; He, LC; Zhang, YY; Han, QD
关键词: cell membrane chromatography ; cell membrane stationary phase. ; alpha(1A) adrenergic receptors ; alpha(1B) adrenergic receptors ; human embryonic kidney 293 cell lines ; immobilization ; liquid chromatography ; drug ; membrane receptor
刊名: ANALYTICAL BIOCHEMISTRY
发表日期: 2005-04-15
DOI: 10.1016/j.ab.2005.01.004
卷: 339, 期:2, 页:198-205
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]: SILICA SURFACE ; RAT AORTA ; SUBTYPES ; RECEPTOR
英文摘要:

As a novel bioaffinity chromatographic technique, cell mermbrane chromatography (CMC) originated in 1996. The cell membrane stationary phase (CMSP) consists of porous silica coated with active cell membranes. By immersing silica into a Suspension of cell membranes, the whole surface of silica was covered by the cell membranes. The present study repeatedly investigated the interaction between ligands and receptors by employing the system of CMC and especially evaltiated the accuracy and feasibility of the CMC model in the study of subtype receptors. The cDNA encoding alpha(1A) or alpha(1B) adrenergic receptors (ARs) was transfected into human embryonic kidney 293 (HEK293) cell lines; cell lines stably overexpressing subtype receptors were obtained. HEK293 alpha(1A) or HEK293 alpha(1B) ARs CMSP were prepared by immobilizing relevant cell membranes on silica. In the described chromatography-based CMSP. the retention times of nine alpha(1) adrenoceptor ligands and calculated capacity factors. as chromatographic parameters, were recorded carefully. These results showed a good correlation with the affinity of the same compounds for the corresponding cloned alpha(1) adrenoceptor subtype. The rank order of capacity factors was consistent with the affinity rank order obtained from radioligand binding assays. The immobilized subtype-selective CMSP was stable and reproducible. The study demonstrates that the HEK293 alpha(1A) and HEK293 alpha(1B) CMSP can be utilized for initial screening of drug candidates. (c) 2005 Elsevier Inc. All rights reserved.

语种: 英语
WOS记录号: WOS:000228192700002
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/54480
Appears in Collections:北京大学第三临床医学院_心血管内科_期刊论文

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作者单位: 1.Xian Jiaotong Univ, Sch Med, Dept Pharmacol, Xian 710061, Peoples R China
2.Xian Jiaotong Univ, Coll Pharm, Xian 710061, Peoples R China
3.Peking Univ, Hosp 3, Inst Vasc Med, Beijing 100083, Peoples R China

Recommended Citation:
Wang, Y,Yuan, BX,Deng, XL,et al. The preparation of HEK293 alpha(1A) or HEK293 alpha(1B) cell membrane stationary phase and the chromatographic affinity study of ligands of alpha(1) adrenoceptor[J]. ANALYTICAL BIOCHEMISTRY,2005,339(2):198-205.
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