Kinetic measurements of molecular interactions by spectrofluorometry.
The kinetics of antibody-antigen interactions are reviewed in terms of general trends observed in both polyclonal and monoclonal antibody populations. Anti-fluorescein antibodies are featured in the review as model proteins to explore fluorescence-based kinetic measurements. Since the fluorescence of the fluorescein ligand is significantly quenched upon interaction with both polyclonal and monoclonal anti-fluorescein antibodies, the quenching parameter can be advantageously employed in measuring the rates of association (k1) and dissociation (k2). The near diffusion-limited k1 rates and the prolonged k2 rates are discussed in terms of antibody affinity and mechanisms involved in ligand binding. Specific prolongation effects of reagents, such as anti-metatype antibodies, on the dissociation rate are discussed in terms of antibody dynamics and conformational substates.
Voss
E W
EW
Department of Microbiology, University of Illinois, Urbana 61801.
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000378:metabolism*; D005452:Fluoresceins; D006241:Haptens; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D013053:Spectrophotometry / Q000379:methods
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D005452:Fluoresceins; D006241:Haptens