CHARACTERIZATION OF A MONOCLONAL-ANTIBODY SPECIFIC FOR H-TYPE-2 STRUCTURE OF ABO BLOOD-GROUP, AND ITS USE FOR MEASURING H-TYPE-2 ON HUMAN RED-BLOOD-CELLS

A monoclonal antibody 3A5 specific for the human red blood cells was p roduced by immunizing BALB/c mouse with human erythrocyte membranes of group O following the immunization protocol of selectively killing th e antigen-stimulated lymphocytes. The monoclonal antibody 3A5 we obtai ned agglutinated red blood cells regardless of individuals of blood gr oup A, B, or O types, but not those from a person with a rare para-Bom bay type which is the H-deficient phenotype. The hemagglutination reac tion of 3A5 was not inhibited by saliva from either secretor or nonsec retor individuals. The specificity of 3A5 was studied by adsorption wi th and elution from synthetic oligosaccharide immunoadsorbents includi ng H disaccharide, and H type 1, 2, 3, and 4 structures. Also, the rea ctivity of 3A5 with synthetic oligosaccharide-BSA complexes for H type 1, H type 2, Le(a), Le(b), A, and B was determined by an immunoassay. We found that 3A5 did not react with any of these synthetic oligosacc harides except H type 2. From these results, the antigenic epitope rec ognized by 3A5 was demonstrated to be the H type 2 structure. Addition ally, the H type 2 substance on erythrocytes was quantitatively analyz ed using 3A5. (C) 1996 Wiley-Liss, Inc.