Comparison of zonal elution and nonlinear chromatography in determination of the interaction between seven drugs and immobilised β2-adrenoceptor
Comparison of zonal elution and nonlinear chromatography in determination of the interaction between seven drugs and immobilised β2-adrenoceptor
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DOI：
10.1016/j.chroma.2015.05.012
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发表时间：
2015-07-03
影响因子：
4.1
通讯作者：
Zhang, Youyi
中科院分区：
文献类型：
--
作者：
Li, Qian;Wang, Jing;Zhang, Youyi
区带洗脱和非线性色谱是亲和色谱中测定药物-蛋白相互作用的两种主流模型。本工作旨在比较区带洗脱法和非线性色谱法在分析7种药物与固定化β（2）-肾上腺素受体（β（2）-AR）相互作用时的结果。区带洗脱结果表明，氯丙那林、克仑特罗、甲氧那明、沙丁胺醇、特布他林、妥洛特罗和班布特罗在固定化β（2）-AR上只有一种结合位点，而非线性色谱证实β（2）-AR与氯丙那林、克仑特罗和班布特罗之间至少存在两种结合位点。在这些位点上，无论是区带洗脱还是非线性色谱，7种药物的缔合常数呈现相同的秩次。与区带洗脱法相比，非线性色谱法计算的缔合常数与放射性配体结合法的相应值具有良好的线性关系。非线性色谱的进样效率明显高于区带洗脱。非线性色谱可能成为高通量测定药物-蛋白质相互作用的有力选择。(C)2015 Elsevier B. V.版权所有。
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 beta(2)-adrenoceptor (beta(2)-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 beta(2)-AR, while nonlinear chromatography confirmed the existence of at least two types of binding sites between beta(2)-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. (C) 2015 Elsevier B.V. All rights reserved.