title: Kinetic titration series with biolayer interferometry.
abstract: Biolayer interferometry is a method to analyze protein interactions in real-time. In this study, we illustrate the usefulness to quantitatively analyze high affinity protein ligand interactions employing a kinetic titration series for characterizing the interactions between two pairs of interaction patterns, in particular immunoglobulin G and protein G B1 as well as scFv IC16 and amyloid beta (1-42). Kinetic titration series are commonly used in surface plasmon resonance and involve sequential injections of analyte over a desired concentration range on a single ligand coated sensor chip without waiting for complete dissociation between the injections. We show that applying this method to biolayer interferometry is straightforward and i) circumvents problems in data evaluation caused by unavoidable sensor differences, ii) saves resources and iii) increases throughput if screening a multitude of different analyte/ligand combinations.
authors.affiliation: Forschungszentrum Jülich, ICS-6 Structural Biochemistry, Jülich, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Frenzel
authors.affiliation: Forschungszentrum Jülich, ICS-6 Structural Biochemistry, Jülich, Germany; Heinrich-Heine-Universität Düsseldorf, Institut für Physikalische Biologie, Düsseldorf, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Willbold
mesh_terms: D007074:Immunoglobulin G; D007368:Interferometry / Q000379:methods*; D007700:Kinetics; D011485:Protein Binding; D020349:Surface Plasmon Resonance
chemical_list: D007074:Immunoglobulin G