基于牛血清白蛋白偶联微球的蛋白-药物结合常数快速测定方法研究

摘要

以氨基化多孔微粒为固相介质，牛血清白蛋白（ BSA）为模型蛋白，通过戊二醛交联反应，制备牛血清白蛋白偶联微球，与药物结合，通过离心沉降微球，高效液相色谱法测定上清液中药物浓度的变化，并代入Rosenthal模型，研发出一种新型方法用于药物蛋白结合常数（ Ka ）的快速测定。利用该方法测定了瑞替加滨与BSA的结合常数，并与传统荧光光谱法对比，两种方法结果一致。键合蛋白微球经PBS缓冲液充分洗脱后，可重复利用。%Amino Porous Particles as the solid medium and bovine serum albumin( BSA )as the Protein model,to PrePare the BSA couPling microsPheres using the couPling agent glutaraldehyde. Combined with drugs through centrifugal sedimentation microsPheres,HPLC method was develoPed for the determination of drug concentration in suPernatant fluid before and after balance,and through Rosenthal model,estab-lished a new and raPid method to determine the binding constant( Ka ). Retigabine and BSA binding con-stant was determined using this method,the result was consistent with the traditional fluorescence titration method. The used BSA couPling microsPheres were eluted with PBS and can be reused.