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目的应用表面等离子共振技术(SPR)建立一种新的测定白蛋白与化合物相互作用的方法。方法将人血清白蛋白(HSA)固定在CM5芯片表面。不同化合物的溶液流经固定于芯片表面的HSA,SPR显示在芯片表面的折射系数变化。分析软件处理数据,用HSA流通池得到的数据减去参比池得到的数据进行拟合,确定结合常数。结果所有测试药物与固定的HSA可逆结合。紫草素1的平衡结合常数KA为3000L.mol-1。SPR样品需要量最小而且能够自动分析,还可以直接检测小分子(Mr308~585)结合,使之适用于评价药物与HSA相互作用。结论HSA主要的药物键合位点没有因为被固定到芯片表面而破坏。验证了SPR可以准确简易测定化合物的结合解离。
Objective To establish a new method for the determination of the interaction between albumin and compounds by surface plasmon resonance (SPR). Methods Human serum albumin (HSA) was immobilized on the surface of CM5 chip. Solutions of different compounds flow through the HSA immobilized on the surface of the chip, and the SPR shows the change of the refractive index at the surface of the chip. The analysis software processes the data, fits the data obtained from the HSA flow cell minus the reference cell, and determines the binding constant. Results All tested drugs reversibly bound to immobilized HSA. Shikonin 1 has an equilibrium binding constant KA of 3000 L · mol -1. SPR samples are of minimal need and can be analyzed automatically. They can also directly detect the binding of small molecules (Mr308 ~ 585) making them suitable for evaluating drug-HSA interactions. Conclusion The main HSA binding site was not damaged by being immobilized to the surface of the chip. It is verified that SPR can accurately and easily determine the compound dissociation.