Individuals who do not possess the D antigen in their red blood cells generate Anti-D antibodies against an antigenic challenge. Prophylaxis with Anti-D immunoglobulin prevents sensitization. The determination of adequate doses of Anti-D in plasma and pharmaceutical products is carried out by radio immuno assay (RIA) and enzyme immuno assay (EIA) or hemagglutination. An in house technique was developed for the quantitation of Anti-D antibodies, as an alternative test to the reference method. It was specific and with good recovery and did not present false positives or autoagglutination. The dose-response curve (mean fluorescence intensity (MFI) versus logarithm of concentration (log C)) was linear (correlation coefficient of 0.99). The method was validated following the criteria of the NIBSC (National Institute of Biological Standards and Control) and the European Pharmacopoeia. Flow cytometry allowed obtaining accurate, precise, sensitive and specific determinations at different concentrations in different biological matrices. The method can be used in highly diluted samples, has a strong fluorescence signal, is simple, fast, reliable and of relatively low cost. Flow cytometry is more efficient than hemagglutination and easier than RIA. With similar security and efficiency standards, it is cheaper than EIA an RIA. This method as a more suitable choice.
Part of the book: Innovations in Cell Research and Therapy