title: Transition towards antigen-binding promiscuity of a monospecific antibody.
abstract: Polyspecificity is defined as the ability of a given antibody molecule to bind a large panel of structurally diverse antigens. A fraction of circulating IgG in all healthy individuals acquires promiscuous antigen-binding activity only after a transient exposure to certain protein destabilizing factors. The molecular mechanisms of this phenomenon are not well understood. Exposures to protein destabilizing agents are common steps in immunoglobulin isolation and purification processes. We performed kinetic and thermodynamic analyses using surface plasmon resonance-based technique in order to characterize the interactions of a single mouse monoclonal antibody to its cognate antigen before and after induction of promiscuous antigen-binding activity. The obtained results, suggest that enhanced antigen binding activity induced by exposure to mild denaturing condition resulted from an increase in the structural flexibility of the antigen-binding site. Further pH and ionic strength-dependence analyses of the antibody/antigen interactions demonstrated that the transition to promiscuous antigen-binding was accompanied by a change in the type of non-covalent forces involved in the complex formation. Moreover, from this study, it is evident that an antibody molecule could use two distinct thermodynamic pathways for binding to the same antigen while retaining the same value of the binding affinity. The obtained results may contribute to the understanding of the molecular mechanisms that lay behind natural antibody polyspecificity.
authors.affiliation: Department of Immunology, Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G.Bonchev St., Block 26, 1113 Sofia, Bulgaria.
authors.forename: Jordan D
authors.initials: JD
authors.lastname: Dimitrov
authors.forename: Sébastien
authors.initials: S
authors.lastname: Lacroix-Desmazes
authors.forename: Srinivas V
authors.initials: SV
authors.lastname: Kaveri
authors.forename: Tchavdar L
authors.initials: TL
authors.lastname: Vassilev
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry; D001666:Binding Sites, Antibody; D051379:Mice; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens