Development and applications of sensitive enzyme immunoassay for antibodies: a review

The sensitivity of the conventional enzyme immunoassay methods is seriously limited by the presence of nonspecific immunoglobulins at high concentrations in test serum. In order to overcome this difficulty, various methods have been developed, and the detection limit of serum antibodies has been lowered to 50-100 ng/l, which is 300,000-fold lower than that by the conventional enzyme immunoassay method using antigen-coated solid phases and anti-immunoglobulin antibody-enzyme conjugates. As a result, anti-insulin antibodies have been demonstrated in most of patients treated with insulin for 0.5-10 months, and anti-thyroglobulin IgG has been demonstrated not only in all patients with Graves' disease and chronic thyroiditis, but also in a large proportion of healthy subjects.