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Specific radioimmunochemical identification and quantitation of hemoglobins a2 and f
Author(s) -
Garver Fred A.,
Jones C. Sidney,
Baker Marilyn M.,
Altay Gultekin,
Barton Betty P.,
Gravely Marsha,
Huisman Titus H. J.
Publication year - 1976
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/ajh.2830010411
Subject(s) - immunogen , hemoglobin , cyanogen bromide , chromatography , chemistry , antiserum , radioimmunoassay , hemoglobin s , thalassemia , agarose , affinity chromatography , microbiology and biotechnology , biochemistry , sickle cell anemia , antibody , biology , immunology , cell , monoclonal antibody , enzyme , peptide sequence , gene , genetics
Hyperimmune antisera to chromatographically purified hemoglobins F and A 2 , 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 A 2 , 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‐A 2 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.