The long-term goal of the proposed research is to develop sensitive, specific and quantitative enzyme-linked immunosorbent assays (ELISA) for (1) measuring anti-RhD (anti-D) antibodies in patients at risk for Hemolytic Disease of the Newborn (HDN), (2) screening individuals who receive anti-D for prophylaxis of HDN, (3) qualitative assessment of anti-D preparations, and (4) use in pharmacokinetic studies. Most clinical laboratories and hospital transfusion services use manual test tube hemagglutination methods to perform anti-D detection and compatibility tests. However, traditional red cell agglutination titering methods have serious limitations. Agglutination reactions are generally detected visually, that is, in these tests, the technician subjectively evaluates the procedure's results and, consequently, antibody detection procedures are slow, labor intensive and error prone. Because of the subjective nature of sample evaluation, results from these tests are far from uniform. In recent years, alternatives to manual test tube methods have been developed. Some of these new technologies have been adapted for large-scale automated testing of blood samples. While automated methods based on agglutination reactions and red cell detection allow for easier manipulation of large numbers of samples, they require several washing and centrifugation steps, and involve very expensive and complex instrumentation. In this grant application, we propose to develop new ELISA assays for determination of anti-D antibody that will be free from the limitations of the conventional agglutination methods. Successful completion of these studies will make available all of the technology needed for a substantial business opportunity to license the technology and manufacture ELISA kits for quantitation of anti-D antibodies and to standardize RhD immune globulin (RhIG) manufacture. ? ?
