高磁场响应性蛋白偶联磁性微球的制备及其在药物-蛋白结合常数测定中的应用  

Protein-coated magnetic beads for the measurement of drug-protein association constant

作　　者：潘晓霞[1] 

机构地区：[1]天津大学药物科学与技术学院,天津300072

出　　处：《应用化工》2014年第8期1531-1535,共5页Applied Chemical Industry

摘　　要：以蛋白偶联磁性微球(MB)为固相基质,通过磁场下快速分离蛋白和上清,高效液相色谱法测定上清中的药物浓度,建立了测定药物与蛋白结合常数(Ka)的新方法。选择对药物非特异性吸附较少和磁响应性较高的四氧化三铁磁性微球为载体,分别以1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐/N-羟基琥珀酰亚胺(EDC/NHS)及戊二醛作为偶联剂,制备牛血清白蛋白(BSA)包覆的磁性微球(MB-BSA),两种偶联方法所得磁性微球的蛋白最大键合量均为50μg/mg。以布洛芬为模型药物,研究药物在蛋白偶联磁性微球上的吸附行为及药物-蛋白的相互作用模式,优化药物吸附和解吸过程。利用高效液相色谱法测定经磁性分离的上清液中的游离药物浓度,通过罗森塔尔方程计算药物与BSA结合常数。最后将所建立的方法应用到色氨酸、安立生坦、瑞替加滨、罗氟司特等4种不同药物与蛋白的结合常数的测定,获得满意结果。A simple and fast method for the determination of the association constant（ Ka ）of drugs to ser-um albumin has been developed by using bovine serum albumin-coated magnetic beads（ MB-BSA）. Opti-mum type of MB which has least nonspecific adsorption effect was screened,the maximum amount of the two protein bond coupling method resulting magnetic microsphere are all 50 μg/mg. The crosslinking method was also optimized to get the appropriate bound capacity of BSA. Then the adsorption equilibrium time and drug-protein interaction mode using ibuprofen as the model drug was studied. Finally,the Ka for ibuprofen,tryptophan,ambrisentan,retigabine and roflumilast were calculated via Rosenthal equation by detecting the concentration of drugs before and after incubation with MB-BSA. Though above research,we improved and perfected the MB-BSA method to calculate the drug-protein association constant.

关 键 词：磁性微球 牛血清白蛋白 药物-蛋白作用 结合常数 

分 类 号：TQ021.9[化学工程] O642[理学—物理化学]