title: Immunoaffinity capillary electrophoresis: determination of binding constant and stoichiometry for antibody-antigen interaction.
abstract: Affinity capillary electrophoresis (ACE) can provide both qualitative and quantitative information on molecular interactions and affords the advantages of very low sample consumption, high mass sensitivity, short analysis time, and the use of automated instrumentation. It has been applied clinically and biochemically to the determination of the binding constant and to the measurement of the binding stoichiometry for interactions between antibodies (Ab's) and antigens (Ag's) in free solution. In many situations, the Ag molecule has two or multiple binding sites, each of which has a similar or different intrinsic affinity for binding independently to the combining site(s) on an Ab molecule. The multivalent binding reactions between Ab and Ag molecules often occur. The objective of this review is to describe the uses of ACE in the determination of binding constants and stoichiometry of Ab-Ag interactions (immunoaffnity capillary electrophoresis), focusing especially on multivalent Ab-Ag interaction modes. Five model binding systems developed recently using ACE techniques are described with principles and examples: (i) divalent mAb-monovalent Ag interaction, (ii) divalent mAb-(homo)polyvalent Ag interaction, (iii) cooperativity of two binding sites of mAb-monovalent Ag interaction, (iv) monovalent Fab-divalent Ag interaction, and (v) polyclonal Ab-monovalent Ag interaction. Finally, the determination of binding stoichiometry of Ab-Ag interactions by ACE is described.
authors.affiliation: Department of Chemistry, National Taiwan University/College of Medicine, 1-1 Jen-ai Road, Taipei, Taiwan, ROC.
authors.forename: Wei-Lung
authors.initials: WL
authors.lastname: Tseng
authors.forename: Huan-Tsung
authors.initials: HT
authors.lastname: Chang
authors.forename: Su-Ming
authors.initials: SM
authors.lastname: Hsu
authors.forename: Ruey-Jien
authors.initials: RJ
authors.lastname: Chen
authors.forename: Shiming
authors.initials: S
authors.lastname: Lin
mesh_terms: D000937:Antigen-Antibody Reactions; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007700:Kinetics