Influence of heavy metal ions on antibodies and immune complexes investigated by dynamic light scattering and enzyme-linked immunosorbent assay.

The effect of Cd2+, Pb2+, Hg2+ and Cu2+ on the aggregation behaviour of monoclonal rat-IgG1-anti-mouse antibodies (kappa-light chain specific) and their antibody-antigen complexes with monoclonal mouse-IgG1 is reported. Investigations were done using the dynamic light scattering method. Cd2+ ions affected the hydrodynamic properties of the antibodies and the immune complex formation very little. More than 4 Cu2+ ions per antibody molecule led to large insoluble aggregates. Pb2+ ions also interacted with antibodies and immune complexes. Instead of "monomeric' antibodies (Ab) or immune complexes (Ab1Ag1), large soluble aggregates were detectable in the solution. Hg2+ ions induced complex formation with 3-4 antibodies per aggregate. Possible kinds of interaction are discussed. Additionally, we tested the antigen binding activity of metal-treated antibodies in ELISA-tests. The Sandwich ELISA technique was used to investigate the serological activity of the metal-treated antibodies, i.e., the reaction with the specific antigen. For these experiments we used the same monoclonal antibodies, mouse-IgG1 and rat-IgG1-anti-mouse. The influence of the above mentioned heavy metal ions was investigated up to a 10-fold molar excess over the antibody concentration. Even at these "unphysiological' high metal ion concentrations an inhibition of the antibody-antigen binding activity was not detectable.