Abstract
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.
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