title: Interpolation method for accurate affinity ranking of arrayed ligand-analyte interactions.
abstract: The values of the affinity constants (kd, ka, and KD) 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 Rmax = 100 RU response level (KD(R100)).
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute, University of Twente, 7500 AB Enschede, The Netherlands; IBIS Technologies, 7521 PR Enschede, The Netherlands. Electronic address: r.b.m.schasfoort@utwente.nl.
authors.forename: Richard B M
authors.initials: RB
authors.lastname: Schasfoort
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute, University of Twente, 7500 AB Enschede, The Netherlands.
authors.forename: Kiki C
authors.initials: KC
authors.lastname: Andree
authors.affiliation: IBIS Technologies, 7521 PR Enschede, The Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: van der Velde
authors.affiliation: IBIS Technologies, 7521 PR Enschede, The Netherlands.
authors.forename: Alex
authors.initials: A
authors.lastname: van der Kooi
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute, University of Twente, 7500 AB Enschede, The Netherlands.
authors.forename: Ivan
authors.initials: I
authors.lastname: Stojanović
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute, University of Twente, 7500 AB Enschede, The Netherlands.
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
mesh_terms: D015374:Biosensing Techniques; D007700:Kinetics; D008024:Ligands; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D008024:Ligands
keywords: Affinity; Biosensor; Kinetics; Label free; SPR imaging