Department of Biopharmacy, Faculty of Pharmacy, Collegium Medicum, Nicolaus Copernicus University, M. Skłodowskiej-Curie 9, 85-094 Bydgoszcz, Poland.
J Pharm Biomed Anal. 2010 Jul 8;52(3):420-4. doi: 10.1016/j.jpba.2009.06.023. Epub 2009 Jun 18.
A simple and fast method for the determination of the association constant (K(a)) of ligand to human serum albumin (HSA) has been developed by using human serum albumin-coated magnetic beads (HSA-MB). To date, magnetic beads (MB) have been increasingly used as a bioseparation tool, especially for DNA, RNA, protein, enzyme and cell isolation or purification. In this study, HSA-MB were used as a new tool to determine the affinity of known ligands to HSA. The K(a) for l-tryptofan, fenoprofen, ketoprofen, tolbutamide and warfarin obtained from Schathard analysis are consistent with previously reported values. The different K(a) values for ketoprofen after the acetylation of HSA-MB by preincubation with acetylosalicylic acid indicate that these beads can be successfully adapted in combined experiment. In addition, the HSA-MB experiment with phenytoin and valproic acid proved to be a simple method to examine drug displacement effect.
一种简单快速的测定配体与人血清白蛋白(HSA)结合常数(K(a))的方法已经被开发出来,该方法使用了人血清白蛋白包被的磁性珠(HSA-MB)。迄今为止,磁性珠(MB)已被越来越多地用作生物分离工具,特别是用于 DNA、RNA、蛋白质、酶和细胞的分离或纯化。在这项研究中,HSA-MB 被用作一种新的工具来测定已知配体与 HSA 的亲和力。通过 Schathard 分析得到的 l-色氨酸、芬布芬、酮洛芬、甲苯磺丁脲和华法林的 K(a)值与之前报道的值一致。HSA-MB 与乙酰水杨酸预孵育进行乙酰化后,酮洛芬的不同 K(a)值表明这些珠粒可以成功地应用于组合实验。此外,苯妥英和丙戊酸的 HSA-MB 实验被证明是一种简单的方法来检查药物置换效应。