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学科主题: 药学
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CCR4 expressing cells cultured adherently on a capillary wall and formaldehyde fixed as the stationary phase for ligand screening by ACE
作者: Yakufu, Pazilaiti1,2; Qi, Hui3,4; Li, Meina1,2; Ling, Xiaomei1,2; Chen, Wenjing1,2; Wang, Ying3,4
关键词: ACE ; CCR4 ; Cell immobilization ; Drug screening ; NLC
刊名: ELECTROPHORESIS
发表日期: 2013-02-01
DOI: 10.1002/elps.201200376
卷: 34, 期:4, 页:531-540
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]: CHEMOKINE RECEPTOR 4 ; PROTEIN-COUPLED RECEPTORS ; NONLINEAR CHROMATOGRAPHY ; AFFINITY-CHROMATOGRAPHY ; FRONTAL ANALYSIS ; T-CELLS ; ELECTROPHORESIS ; BINDING ; CZE
英文摘要:

We have developed an on-line screening method for CC chemokine receptor 4 (CCR4) ligands, in which the whole cells expressed with CCR4 were cultured adherently and immobilized on the inner wall of the capillary as the stationary phase for the first time. Moreover, in this method it is unnecessary to isolate and purify the target receptors from cell membranes. Therefore, it is possible to almost completely preserve the native conformation of the target receptors. The binding activities of the immobilized CCR4 did not change. A known antagonist of CCR4, compound A, was employed to validate the bioactivity of the cell layer and stability of this method. The intraday, interday, and batch-to-batch reproducibilities were investigated (RSD 13.9%). Nonlinear chromatography was used to calculate the binding constant between the compound A and CCR4 (6.4 x 104/M, RSD = 4.96%). Using this method, the qualitative and quantitative characterizations of 23 computer-aided drug design compounds were achieved and the kinetic parameters (K, ka, kd, and k) were obtained by nonlinear chromatography. Three active compounds were screened out, which also showed activity in chemotaxis inhibition assay. The experimental results show that this method is simple, sensitive, and efficient for drug screening. Moreover, it offered a novel way to detect the nonspecific interactions between ligands and cell membrane.

语种: 英语
所属项目编号: 30872292 ; 90813025 ; 81072612 ; 7102107 ; K20110109 ; 20110001110021
项目资助者: National Natural Science Foundation of China ; Natural Science Foundation of Beijing ; Open Foundation of State Key Laboratory of Natural and Biomimetic Drugs ; Specialized Research Fund for the Doctoral Program of Higher Education of China
WOS记录号: WOS:000315169800008
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内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/65520
Appears in Collections:北京大学药学院_期刊论文

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作者单位: 1.Peking Univ, State Key Lab Nat & Biomimet Drugs, Sch Pharmaceut Sci, Beijing 100871, Peoples R China
2.Peking Univ, Dept Pharmaceut Anal, Sch Pharmaceut Sci, Beijing 100871, Peoples R China
3.Peking Univ, Ctr Human Dis Genom, Beijing 100871, Peoples R China
4.Peking Univ, Sch Basic Med Sci, Dept Med Immunol, Beijing 100871, Peoples R China

Recommended Citation:
Yakufu, Pazilaiti,Qi, Hui,Li, Meina,et al. CCR4 expressing cells cultured adherently on a capillary wall and formaldehyde fixed as the stationary phase for ligand screening by ACE[J]. ELECTROPHORESIS,2013,34(4):531-540.
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