Interference of immunoglobulins in the radioimmunoassay of human beta-endorphin.

Abstract

The interference of immunoglobulins in the radioimmunoassay (RIA) of human beta-endorphin was investigated. Human IgM showed no cross-reactivity. Human IgA showed a weak cross-reaction, but the dilution curve of IgA did not show parallelism with the standard curve of beta-endorphin, thus indicating its antigenic difference. The dilution curves of human IgG showed 0.18% displacement with respect to the human beta-endorphin standard curve, with good parallelism. Moreover, five patients with multiple myeloma of the IgG type showed falsely elevated beta-endorphin levels. We investigated the possibility that certain IgGs may be responsible for the displacement of [125I]beta-endorphin in the beta-endorphin kit. The apparent beta-endorphin level of plasma from multiple myeloma patients was markedly decreased after affinity chromatography of the serum on protein A-Sepharose. In another 3 patients with multiple myeloma, we examined IgG interference by measuring the beta-endorphin levels in their lyophilized IgG diluted with saline. The results demonstrated high values of 20.2, 25.5 and 21.2 pmol/l respectively, also showing good parallelism. These immunological parallels to human beta-endorphin verify that a part of the amino acid sequence of human IgG is similar to that of human beta-endorphin. Consequently, in the measurement of beta-endorphin with polyclonal antibody, the results may sometimes be spuriously high due to cross-reaction with IgG, e.g., in patients with IgG myeloma. To avoid IgG interference, a specific monoclonal antibody to synthetic beta-endorphin should be used rather than polyclonal antibodies.