Characterization of a murine monoclonal antibody specific for the human P1 blood group antigen.
Four monoclonal antibodies (MAbs) directed against the P1 blood group antigen were produced by hybridomas obtained from mouse immunized with turtle-dove avomucoid. One of the MAb (154 IX B6) selected as a blood typing reagent agglutinated native P1 and Pk1 red cells with a high titer but was inactive against native P2, Pk2 and p erythrocytes. After papain treatment the reactivity towards P1 and Pk1 erythrocytes was enhanced whereas p erythrocytes remained unreactive. A weak cross-reactivity of the MAb with the Pk antigen was suspected since enzyme-treated Pk2 erythrocytes became significantly agglutinated. Further analysis of the antibody specificity was established by binding studies using neutral glycolipids prepared from P1 and P2 erythrocytes, affinity immunoabsorbents carrying known oligosaccharide structures and hapten inhibition with synthetic oligosaccharides. The MAb bound weakly to the Gal alpha 1-4Gal structure common to P1 and Pk antigens but had a marked preference for the P1 determinant (Gal alpha 1-4 Gal beta 1-4 GlcNAc) and the binding was abolished by prior treatment of oligosaccharide antigens by alpha(not beta)-galactosidase, which supports evidence that a terminal alpha-galactose residue is involved in the blood group P1 and Pk specificities. The MAb has a slightly broader specificity than the human anti-P1 counterpart but can be used safely for routine blood typing.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000502:physiology*; D000918:Antibody Specificity; D001789:Blood Group Antigens / Q000276:immunology*; D001788:Blood Grouping and Crossmatching / Q000379:methods; D002855:Chromatography, Thin Layer; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D007519:Isoantigens / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D010127:P Blood-Group System / Q000276:immunology*; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D001789:Blood Group Antigens; D007519:Isoantigens; D010127:P Blood-Group System