Use of a linear gradient flow program for liquid chromatography-mass spectrometry protein-binding studies

A rapid screening method to measure drug-protein binding using an immobilized human serum albumin (HSA) column was developed. This method utilizes a linear gradient flow-rate to accelerate the elution of strong binders to the HSA column. Post-column addition of a pressure relief valve enables mass spectrometric detection at relatively high mobile phase flow-rates (i.e., 2 ml/min).