Is bivalent binding of monoclonal antibodies to different antigenic areas on the hemagglutinin of influenza virus required for neutralization of viral infectivity?

Biological activities of Fab fragments of monoclonal IgG antibodies to each of four nonoverlapping antigenic areas on the hemagglutinin molecule of A/seal/Massachusetts/1/80 (H7N7) influenza virus were examined. Fab fragments of the antibodies belonging to groups I and II neutralized viral infectivity. These Fab fragments inhibited hemagglutination of the virus and virus-induced hemolysis at pH 5.9. On the other hand, Fab fragments of groups III and IV antibodies showed neither neutralization nor hemolysis-inhibition activities, while intact IgG molecules of groups III and IV effectively neutralized viral infectivity and inhibited virus-induced hemolysis, as previously found. These IgG molecules scarcely or did not inhibit hemagglutination of the virus. Neutralization of viral infectivity, however, was observed when the virus was coated with Fab fragments of groups III and IV antibodies and then incubated with anti-Fab fragment antibodies. These findings suggest that bivalent binding of the IgG antibodies of groups III and IV is required for neutralization of viral infectivity through a proposed mechanism by which these antibodies interfere with a low pH-induced conformational change resulting in inhibition of the fusion step of the viral replication process.