Measurements of drug-protein binding by using immobilized human serum albumin liquid chromatography-mass spectrometry.

An HPLC/MS based method was used for fast and convenient determination of drug plasma-protein interactions in early drug discovery screening by employing a human serum albumin affinity column. Results from this methodology were compared with data from ultrafiltration or dialysis methods, and good agreement was observed. A compound not suitable for ultrafiltration due to the very high non-specific binding to artificial membrane of ultrafiltration device was also successfully analyzed by this method, and the protein binding determined by this chromatography method was very similar to data obtained by dialysis technique employing biological membranes. The immobilized HSA column LC/MS method also proved to be more reproducible and precise compared to ultrafiltration method in drug protein binding measurements.