A monoclonal antibody-linked immunoassay for hemoglobin H disease

Summary A murine monoclonal antibody (mAb) was generated that recognizes hemoglobin (Hb) H, the tetrameric form (₄) of human -globin chains. The antibody ₄-1 (1, ) does not react with Hbs A, F, Bart's, or isolated chains, indicating that the antibody recognizes an epitope comprised of multiple chains. A simple, rapid, and sensitive enzyme immunoassay was established to detect and quantitate Hb H in hemolysates from subjects with Hb H disease. The globin level in these patients was also measured using the monoclonal antibody -1, which is specific for chains of Hb A₂. With these assays, 20 hemolysates from subjects with Hb H disease' ten from normal adults and ten from newborn babies were analyzed. The percent of Hb H ranged from 1.5% to 25% in Hb H patients. There was a significant average reduction (32%) in chains in these samples as compared with the normal average adult value. The decreased expresion of chains thus results in a reduction of the levels of normal Hbs A and A₂ and accumulation of ₄, causing Hb H disease.Work supported by NIH grant HLB-41544