Human polyreactive and monoreactive antibodies: effect of glycosylation on antigen binding.

The present experiments were initiated to determine whether the carbohydrate portions of antibody molecules contribute to polyreactivity. Cell lines making human monoclonal polyreactive or monoreactive antibodies of the immunoglobulin (Ig) M, IgG and IgA isotypes were treated with tunicamycin to block N-linked glycosylation of the proteins. Analysis of the secreted native and non-glycosylated proteins revealed a > 95% inhibition of [3H]mannose incorporation. Electrophoresis on sodium dodecyl sulphate-polyacrylamide gels of the proteins from tunicamycin-treated cells showed increased mobility and the absence of [3H]mannose incorporation of the immunoglobulin heavy chains, consistent with the lack of glycosylation. The native and non-glycosylated antibodies were then tested for their ability to bind different antigens. Despite the lack of glycosylation, both polyreactive and monoreactive antibodies bound to antigens with little if any loss of reactivity or specificity. It is concluded that the carbohydrate moieties do not contribute significantly to polyreactivity.