Interference in immunoenzymometric assays caused by IgM anti-mouse IgG antibodies.

ABSTRACT

Serum samples from a female patient gave falsely elevated results in nine of ten immunoenzymometric assays (IEMAs) that used mouse monoclonal antibodies specific for human chorionic gonadotropin (n = 8), thyrotropin, or creatine kinase-MB isozyme. In contrast, normal results were obtained in five radioimmunoassays that used either mouse monoclonal antibodies or antisera specific for human chorionic gonadotropin (n = 3) or thyrotropin (n = 2). Incubation of her serum with IgG from different species or with F(ab)'2 from mouse IgG prior to IEMA showed that the interference was markedly inhibited by mouse IgG, indicating an antibody specific for the Fc portion of mouse IgG. The interfering activity was bound to and eluted from a column containing Protein A-Sepharose CL-4B. Fractionation of the eluted protein over another column containing Sephacryl S300 showed the activity was enriched in the first protein peak, which contained predominantly IgM. A model is proposed to explain how IgM anti-mouse IgG antibody selectively interferes in IEMAs that use mouse monoclonal antibodies.