title: Biointeraction analysis of immobilized antibodies and related agents by high-performance immunoaffinity chromatography.
abstract: A method is described based on high-performance immunoaffinity chromatography for examining the interactions of immobilized antibodies or related binding agents with their targets. It is shown how this method can be used to obtain information on the binding, elution and regeneration kinetics of immobilized binding agents, such as those used with immunoaffinity supports. The theory behind this approach is briefly described and it is demonstrated how both the kinetic and thermodynamic properties of a biointeraction can be determined experimentally through this method. Several applications are used to illustrate this technique, including antibody-antigen interactions and the binding of aptamers with their targets in the presence of silica-based supports. The same approach can be adapted for use with other types of targets, binding agents and support materials.
authors: Chemistry Department, University of Nebraska-Lincoln, Lincoln, NE 68588 0304, USA.
authors: Erika
authors: E
authors: Pfaunmiller
authors: Annette C
authors: AC
authors: Moser
authors: David S
authors: DS
authors: Hage
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000737:chemistry; D000327:Adsorption; D055910:Antibodies, Immobilized / Q000032:analysis* / Q000737:chemistry; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000737:chemistry; D000941:Antigens / Q000032:analysis / Q000737:chemistry*; D001280:Atrazine / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D013816:Thermodynamics
chemical_list: D055910:Antibodies, Immobilized; D000936:Antigen-Antibody Complex; D000941:Antigens; D008024:Ligands; D015084:2,4-Dichlorophenoxyacetic Acid; D001280:Atrazine