title: Reevaluation of stoichiometry and affinity/avidity in interactions between anti-hapten antibodies and mono- or multi-valent antigens.
abstract: In order to obtain further information on the interaction between antigens (Ags) and B cell Ag receptors (BCR) for a better understanding of the relationship between signals resulting from Ag binding and B cell activation, effects of Ag valence and size on the apparent association constant, i.e. the avidity as well as the molecular stoichiometry of immune complexes in Ag-antibody (Ab) interactions were studied. Hapten conjugates using proteins of various molecular weights, such as hen egg lysozyme (HEL), ovalbumin (OVA), bovine serum albumin (BSA), and chicken gammaglobulin (CGG), were prepared for this purpose. Different ratios of the hapten, (4-hydroxy-3-nitrophenyl)acetyl (NP), to the protein were used for conjugation, and interactions between anti-NP monoclonal Abs (mAbs) and the NP conjugates were evaluated by surface plasmon resonance. It was founded that the two binding sites of an Ab were able to simultaneously accommodate two NP(1)-HEL, resulting in a tri-molecular complex, Ag(2)Ab(1). However, NP conjugates of the higher-molecular-weight proteins, OVA and BSA, formed only Ag(1)Ab(1), irrespective of hapten valence. This was thought to be due to steric hindrance caused by the binding of the first Ag. These results suggested that the stoichiometry depended largely on the size of the Ag involved and that mAbs with a low affinity are more efficient at raising the binding strength through divalent interaction since the avidity of two mAbs in interactions with highly haptenated BSA was not significantly different in spite of a 10-fold difference in affinity to the monovalent NP(1)-HEL.
authors.affiliation: Research Institute for Biological Sciences (RIBS), Science University of Tokyo, 2669 Noda, Chiba 278-0022, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Oda
authors.forename: T
authors.initials: T
authors.lastname: Azuma
mesh_terms: D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology*; D006241:Haptens / Q000276:immunology*; D018448:Models, Immunological; D009113:Muramidase / Q000276:immunology; D009596:Nitrophenols / Q000276:immunology*; D010047:Ovalbumin / Q000276:immunology; D010648:Phenylacetates; D012710:Serum Albumin, Bovine / Q000276:immunology; D020349:Surface Plasmon Resonance; D013816:Thermodynamics; D005719:gamma-Globulins / Q000276:immunology
chemical_list: D000941:Antigens; D006241:Haptens; D009596:Nitrophenols; D010648:Phenylacetates; D005719:gamma-Globulins; C026242:4-hydroxy-5-nitrophenyl acetic acid; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D009113:Muramidase