A protein-coated magnetic beads as a tool for the rapid drug-protein binding study.
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.
Marszałł
Michał Piotr
MP
Department of Biopharmacy, Faculty of Pharmacy, Collegium Medicum, Nicolaus Copernicus University, M. Skłodowskiej-Curie 9, 85-094 Bydgoszcz, Poland. mmars@cm.umk.pl
Buciński
Adam
A
D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008280:Magnetics; D008863:Microspheres; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*
D002352:Carrier Proteins; D008024:Ligands; D004364:Pharmaceutical Preparations; D012709:Serum Albumin