Rapid analysis of the interactions between drugs and human serum albumin (HSA) using high-performance affinity chromatography (HPAC)
Rapid analysis of the interactions between drugs and human serum albumin (HSA) using high-performance affinity chromatography (HPAC)
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DOI：
10.1016/j.jchromb.2008.05.029
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发表时间：
2008-07-01
影响因子：
3
通讯作者：
Wainer, Irving W.
中科院分区：
文献类型：
--
作者：
Kim, Hee Seung;Wainer, Irving W.
本研究在固定化人血清白蛋白(HSA)高效亲和层析(HPAC)柱上采用区带洗脱和正面亲和层析相结合的方法，考察了平衡透析和超滤研究的各种化合物的结合常数。用区带洗脱层析法测定参比化合物的保留因子，用前向亲和层析法测定参比化合物的结合常数。保留因子与参比化合物的缔合常数之间存在良好的线性关系(r(2)=0.9993)。这个标准图谱后来被用于快速测定各种药物的结合常数，这些药物对人血清白蛋白显示出低到中等的结合亲和力。将这些药物的结合常数与文献中更常用的方法(即平衡透析或超滤)进行比较，得到了相对较高的相关值(r(2)=0.945)。这种区带洗脱和前沿亲和层析相结合的测定结合常数的方法与传统方法相比显示出一些优点。根据感兴趣的药物的不同，药物与人血清白蛋白的结合常数可以快速测量到1.5分钟。其他显著的优点包括易于自动化，以及同时区分手性化合物的缔合常数的能力。同样的方法可以用于研究其他药物和蛋白质的相互作用，并应进一步改善整体药物筛选过程。爱思唯尔出版公司(Elsevier B.V.)
This study used a combination of zonal elution and frontal affinity chromatography on immobilized human serum albumin (HSA) high-performance affinity chromatography (HPAC) column to examine the association constants of various compounds that have been studied by equilibrium dialysis or ultra filtration. A standard plot was generated from retention factors of reference compounds using zonal elution chromatography against association constants of reference compounds using frontal affinity chromatography. The linear relationship was established (r(2) = 0.9993) between retention factors and association constants of reference compounds. This standard plot was later used for rapid determination of association constants of various drugs which show low to medium binding affinity to HSA. Association constants of those drugs from this study were compared to that of more generally used methods (i.e., equilibrium dialysis or ultra filtration) from literature and resulted in a relatively high correlation (r(2) = 0.945) value. This combination of zonal elution and frontal affinity chromatography method for determining association constants showed several advantages against traditional methods. Depending on drugs of interest, an association constant of drug to HSA can be measured as fast as 1.5 min. Other notable advantages include an ease of automation and its ability to distinguish association constants of chiral compounds at the same time. The same approach could be used for studying interaction of other drugs and proteins and should further improve overall drug screening process. Published by Elsevier B.V.