title: Synergistic monoclonal antibodies' interactions and their use for determination of antibody specificities.
abstract: Three monoclonal antibodies (MAb 3C11, F11 and 10D6) to human growth hormone (hGH) recognize independent epitopes and show mutually enhancing properties. Thus, 125I-hGH binding to each of these MAb augmented significantly in the presence of each one of the other two MAb. Moreover, preincubation of the hormone with paired MAb gave rise to ternary complexes (Ag:Ab1:Ab2) which bound better than the free tracer to the third MAb previously captured on a solid-phase. Highly stable quaternary complexes (Ag:Ab1:Ab2:Ab3) were thus formed. Since Fab fragments from the three MAb displayed the same behavior as the whole Ab molecule, neither the formation of multimolecular cyclic complexes nor the occurrence of interactions through Fc fragments could explain the reciprocal MAb binding enhancement. Therefore, the results obtained suggest that MAb 3C11, F11 and 10D6 produce some modification in the Ag, each one improving the binding of the two other MAb. Additionally, the inhibition of the formation of quaternary complexes between the MAb and hGH was used to evaluate specific Ab populations in polyclonal antisera, avoiding the masking effect of enhancing Ab. The results obtained indicate that Ab directed to the hGH antigenic domains defined by MAb 3C11, F11 and 10D6 could be detected in spite of the presence of enhancing Ab to all three MAb.
authors.affiliation: Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
authors.forename: K A
authors.initials: KA
authors.lastname: Gómez
authors.forename: L A
authors.initials: LA
authors.lastname: Retegui
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D013006:Growth Hormone / Q000276:immunology*; D006801:Humans; D011487:Protein Conformation
chemical_list: D000911:Antibodies, Monoclonal; D013006:Growth Hormone