| The objective of the study was to prepare the monoclonal antibody (mAb) with universal reactivity against human erythrocytes and identify its properties for the construction of bispecific antibodies (BsAb) detected further antibodies or antigens in whole blood.The hybridoma was isolated with conventional technology using spleen cells from BALB/c mice immunized with the membrane antigens of human type O erythrocyte. The antibodies were screened by hexadimethrine bromide (polybrene) test and slide hem-agglutination assay. The titer, stability, affinity, biological activity, as well as other properties of the obtained hybridoma cells and mAb were also identified.The result showed that the hybridoma cell named 2E8 (mAb2E8) secreted non-agglutinating antibodies-IgG1. The titers of culture supernatant and ascites of hybridoma cells were 1 : 1024 and 1 : 6.4×107 respectively. mAb2E8 had no species or blood group cross-agglutination reaction. When the affinity of mAb2E8 was evaluated by agglutination reaction, the erythrocyte initiated to agglutinate within 7 seconds and the clot exceeded 1mm2 in 3 minutes. When red cells were sensitized with mAb2E8, the binding and phagocytosis of red cells by monocytes and the lysis of red cells by monocytes or lymphocytes were great, the hemolyses of red cells by alexin was great too. Moreover, in the process of screening antibodies, we employed hexadimethrine bromide (Polybrene) test, which was used for the first time to prepare the mAb in the world.The non-agglutinating mAb 2E8 with universal reactivity against RBC was prepared successfully. The results would be useful for preparation of BsAb for the detection of antibodies or antigens in human whole blood samples, as well as for the potential use of erythrocyte-based treatment.
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