Development of affinity selection mass spectrometry methods

National Center For Advancing Translational Sciences

Investigators

Abstract

This project is to develop an ASMS protocol using either newly developed beads from Sciex or with the commercially available paramagnetic beads. For both beads, the workflow is the same. We first attach the protein to the bead and then isolate the small molecules that bind to the protein. We use mass spectrometry to determine the identity of those small molecules that bound to the protein and were isolated. Upcoming work involves: Perform a saturation assay where the protein target is incubated with increasing concentration of ligand and the MS response is related to the concentration of L-P complex. Kd = [P][L]/[LP]. This is the Binding Fraction Analysis (BFA). The ligand concentration at which the MS response reaches half of the maximum signal is referred to as the apparent Kd.