Specific radioimmunochemical identification and quantitation of hemoglobins a2 and f

Hyperimmune antisera to chromatographically purified hemoglobins F and A2, were produced in rabbits and made specific for the immunogen by adsorption with normal human hemoglobin A conjugated to cyanogen bromide‐activated agarose. A radioimmunoassay was established that permitted identification and quantitation of each of these two minor hemoglobins in hemolysates containing other hemoglobin components. The quantities of hemoglobins A2, and/or F present in hemolysates of individuals with β‐thalassemia, sickle cell anemia, Hb‐C disease, and other hematological disorders were determined immunochemically, and the results were compared to values obtained by microcolumn chromatography for the measurement of Hb‐A2 or with the alkali denaturation technique in quantitating Hb‐F. The immunoassay procedure has a greater sensitivity than other commonly employed techniques and can detect as little as 0.05 μg of these hemoglobins.