Enzyme and DTT treatment of adherent RBCs for antibody identification by a solid phase immunoassay system

Abstract

Treatment of RBCs with protease enzymes or dithiothreitol (DTT) causes denaturation of several RBC antigens and is regularly used in antibody identification. In this study, we have standardized enzyme and DTT treatment of adherent RBCs in the magnetic-mixed passive hemagglutination assay (M-MPHA) for antibody identification. We have also tried drying these treated RBCs. The optimal enzyme and DTT treatment conditions for intact adherent RBCs were determined, in addition to the optimal condition for drying RBCs. All tested agents with the exception of chymotrypsin could be applied to RBC drying, but the optimal condition for drying was different from that for intact RBCs. Enzyme or DTT treatment of adherent RBCs and their application in M-MPHA provide a simple and convenient method for antibody identification. Furthermore, the technique of drying treated RBCs provides an even stronger antibody identification tool, since dried RBCs can be stored for a long period.