A Two-Stage Immunoradiometric Assay with125I-Staphylococcal Protein A for the Detection of Antibodies and Complement on Human Blood Cells

Abstract

A sensitive two‐stage immunoradiometric assay (IRMA) using unlabeled specific antiglobulin sera followed by binding of radiolabeled staphylococcal protein A has been developed for detection of human IgG, IgM, IgA and complement C3 on human blood cells. In an experimental model system, purified human immunoglobulins (Ig) were coupled onto red blood cells, platelets, and leukocytes. The Ig‐coated red blood cells were analyzed in parallel by three methods: the radioimmune antiglobulin test with radiolabeled anti‐Ig; the radiolabeled staphylococcal protein A test, and the IRMA. Of the three methods evaluated, the latter was found to be sensitive and the easiest to perform. The applicability of the IRMA was established by investigating a group of 14 selected patients with autoimmune warm type hemolytic anemia. We observed that the IRMA detected cell‐bound antibodies (IgG and/or IgM) in several cases where conventional assays yielded negative results.