A protein-coated magnetic beads as a tool for the rapid drug-protein binding study

摘要

A simple and fast method for the determination of the association constant (Ka) 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 Ka for L-tryptofan, fenoprofen, ketoprofen, tolbu-tamide and warfarin obtained from Schathard analysis are consistent with previously reported values. The different Ka values for ketoprofen after the acetylation of HSA-MB by preincubation with acetylosali-cylic 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.