title: Comparison of zonal elution and nonlinear chromatography in determination of the interaction between seven drugs and immobilised β(2)-adrenoceptor.
abstract: Zonal elution and nonlinear chromatography are two mainstream models for the determination of drug-protein interaction in affinity chromatography. This work intended to compare the results by zonal elution with that by nonlinear chromatography when it comes to the analysis of the interaction between seven drugs and immobilised β2-adrenoceptor (β2-AR). The results of the zonal elution showed that clorprenaline, clenbuterol, methoxyphenamine, salbutamol, terbutaline, tulobuterol and bambuterol have only one type of binding site on immobilised β2-AR, while nonlinear chromatography confirmed the existence of at least two types of binding sites between β2-AR and clorprenaline, clenbuterol and bambuterol. On these sites, both zonal elution and nonlinear chromatography presented the same rank order for the association constants of the seven drugs. Compared with the data from zonal elution, the association constants calculated using nonlinear chromatography gave a good linear response to the corresponding values by radio-ligand binding assay. The sampling efficiencies of nonlinear chromatography were clearly higher than zonal elution. Nonlinear chromatography will probably become a powerful alternative for the high throughput determination of drug-protein interaction.
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Xi'an Innovation College of Yan'an University, Xi'an 710100, China.
authors.forename: Yuqing Yuan
authors.initials: YY
authors.lastname: Zheng
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Lingjian
authors.initials: L
authors.lastname: Yang
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Liujiao
authors.initials: L
authors.lastname: Bian
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Institute of Vascular Medicine, Peking University, Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing 100083, China.
authors.forename: Zijian
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an 710069, China. Electronic address: zhaoxf@nwu.edu.cn.
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Institute of Vascular Medicine, Peking University, Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing 100083, China. Electronic address: zhangyy@bjmu.edu.cn.
authors.forename: Youyi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000420:Albuterol / Q000737:chemistry; D000438:Alcohols / Q000737:chemistry; D001665:Binding Sites; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002846:Chromatography, Affinity; D002976:Clenbuterol / Q000737:chemistry; D004347:Drug Interactions; D015725:Fluconazole / Q000737:chemistry; D011869:Radioligand Assay; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry*; D013726:Terbutaline / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D000438:Alcohols; D018343:Receptors, Adrenergic, beta-2; C010809:tulobuterol; D015725:Fluconazole; D013726:Terbutaline; D000420:Albuterol; D002976:Clenbuterol; C047766:bambuterol
keywords: Affinity chromatography; Drug–protein interaction; Nonlinear chromatography; Zonal elution; β(2)-Adrenoceptor