title: Bivalent kinetic binding model to surface plasmon resonance studies of antigen-antibody displacement reactions.
abstract: Molecular and functional analysis of small molecule binding to protein can provoke insights into cellular signaling and regulatory systems as well as facilitate pharmaceutical drug discovery. In label free small molecule detection the displacement assay format can be applied. This is beneficial because displacement of high molecular weight receptors is detected instead of low molecular weight ligand as in classical binding analysis. Thus, detection limit is potentially lowered. Using the influenza haemagglutinin (HA) peptide binding to mono or bivalent anti-haemagglutinin peptide antibody displacement assay formats could be established. The exact time resolved analysis of binding and dissolution of ligand HA and anti-Haemagglutinin peptide antibody was achieved with surface plasmon resonance (SPR) spectroscopy. Mathematical models could be developed from kinetic equations of ligand binding to mono or bivalent antibodies. With this, an accurate simulation of the SPR results was reached. The simulation plot had to be exactly adjusted to the SPR results to determine all kinetic rate constants defining ligand and receptor binding kinetics. Large variations in receptor concentration gave almost identical rate constants in binding. It became obvious that rebinding is in any case not necessary to understand the binding kinetics of our model system HA/anti-HA. Maximum decline of SPR response could be used to determine ligand concentrations in analyte.
authors.affiliation: Institute for Materials Science and Max Bergmann Centre of Biomaterials, Technische Universität Dresden, 01062 Dresden, Germany.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Gelinsky-Wersing
authors.affiliation: Siemens AG, Corporate Technology, 81739 München, Germany. Electronic address: wolfram.wersing@t-online.de.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Wersing
authors.affiliation: Institute for Materials Science and Max Bergmann Centre of Biomaterials, Technische Universität Dresden, 01062 Dresden, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Pompe
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry* / Q000276:immunology; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry* / Q000276:immunology; D000914:Antibodies, Viral / Q000737:chemistry* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D008956:Models, Chemical; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D018033:Antibodies, Bispecific; D058846:Antibodies, Monoclonal, Murine-Derived; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C483468:hemagglutinin, human influenza A virus
keywords: Kinetic rate constants; Kinetic simulation; Ligand binding; Protein interaction; SPR detection; Small molecule sensor