Investigations using a novel monoclonal antibody to the glycosylphosphatidylinositol-anchored protein that carries Gregory, Holley, and Dombrock blood group antigens (original) (raw)
The high-frequency Hy and Gy" anti ens have been shown to reside monoclonal antibod 5g10, was produced with specificity related to the human Grego S t u d y k l g n and Methods: The a n t i b reacted in direct hemagglutination assays, and its specificity was investigated by radioimmunoassay, inhibition assay, and Western blotting. Results: The 5810 antibody failed to bind to abnormal paroxysmal nocturnal hemoglobinuria red cells and human erythroleukemia cell line K562, but it was weakly reactive with HEL cells. Red cells, but not other circulating hematopoietic cells, express the 581 0 antigen. The 581 0 antibody had a specificity similar but not identical to that of Gy. Gy(a-) Hy-negative red cells reacted extremely weakly with 5810 antibody, but Gy(a-) Hy-negative red cells treated with a variety of proteases bound 5810 antibody strongly. This suggests that these cells express a variant form of the protein recognized by 581 0. Concluslon: Identification of a monoclonal antibody to this glycosylphosphatidylinositol-linked protein opens a new avenue for investigation of the biochemistry, g e netics, and function of the lycosylphosphatidylinositol-linked protein that bears the Gy, Hy, and Do antigens.%RANSFUSIDN 1995;35:459464. on the same protein. G (a-) Hy-negative red cel B s are also Do(a-b-). A mouse Holley, andYbombrock blood roup antigens. Abbrevlatlons: DTT = dlthlothmltol; GPI = glycorylphorphatMyllnoritol; MAIM = monoclonal antlbodyapeclfk lmmoblllzatlon oferythrocyteantlgens; MoAb = mone clonal antibody; PBS = phosphate-buffered saline; RBC(8) = red mll(a); RIA = mdlolmmunoassey; TBS =Trls-buffered saline. SEVERAL HIGH-FREQUENCY human blood group antigens are known to reside on glycosylphosphatidylinositol (GP1)-anchored red cell (RBC) membrane proteins.' The
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