title: Analyzing a kinetic titration series using affinity biosensors.
abstract: The classical method of measuring binding constants with affinity-based biosensors involves testing several analyte concentrations over the same ligand surface and regenerating the surface between binding cycles. Here we describe an alternative approach to collecting kinetic binding data, which we call "kinetic titration." This method involves sequentially injecting an analyte concentration series without any regeneration steps. Through a combination of simulation and experimentation, we show that this method can be as robust as the classical method of analysis. In addition, kinetic titrations can be more efficient than the conventional data collection method and allow us to fully characterize analyte binding to ligand surfaces that are difficult to regenerate.
authors.affiliation: Biacore AB, Uppsala, Sweden.
authors.forename: Robert
authors.initials: R
authors.lastname: Karlsson
authors.forename: Phinikoula S
authors.initials: PS
authors.lastname: Katsamba
authors.forename: Helena
authors.initials: H
authors.lastname: Nordin
authors.forename: Ewa
authors.initials: E
authors.lastname: Pol
authors.forename: David G
authors.initials: DG
authors.lastname: Myszka
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism; D000915:Antibody Affinity; D015374:Biosensing Techniques; D007700:Kinetics; D010770:Phosphotransferases / Q000378:metabolism; D017430:Prostate-Specific Antigen / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding; D012984:Software; D019311:Staurosporine / Q000378:metabolism; D017186:Titrimetry
chemical_list: D000911:Antibodies, Monoclonal; D010770:Phosphotransferases; D017430:Prostate-Specific Antigen; D019311:Staurosporine