Interpolation method for accurate affinity ranking of arrayed ligand-analyte interactions

The values of the affinity constants (k(d), k(a), and K-D) that are determined by label-free interaction analysis methods are affected by the ligand density. This article outlines a surface plasmon resonance (SPR) imaging method that yields high-throughput globally fitted affinity ranking values using a 96-plex array. A kinetic titration experiment without a regeneration step has been applied for various coupled antibodies binding to a single antigen. Globally fitted rate (kd and ka) and dissociation equilibrium (KD) constants for various ligand densities and analyte concentrations are exponentially interpolated to the KD at R-max = 100 RU response level (K-D(R100)). (C) 2016 Elsevier Inc. All rights reserved.