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Monoclonal antibody‐based methods for quantitation of hemoglobins: application to evaluating patients with sickle cell anemia treated with hydroxyurea
Author(s) -
Epstein Nava,
Epstein Michael,
Boulet Arielle,
Fibach Eitan,
Rodgers Griffin P.
Publication year - 1996
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.1996.tb00484.x
Subject(s) - monoclonal antibody , horseradish peroxidase , hemoglobin , microbiology and biotechnology , antibody , sickle cell anemia , flow cytometry , immunolabeling , chemistry , monoclonal , red blood cell , anemia , enzyme , cell , biology , biochemistry , immunology , medicine , immunohistochemistry
High‐titer monoclonal antibodies (mAb) were raised against chromatographically purified human hemoglobin (Hb) species. These mAb were specific for either Hb A, Hb F, Hb S or Hb C. Based on these antibodies, which were directly conjugated with either fluorochromes or an enzyme (horseradish peroxidase), we developed immunoassays for determining the Hb profile in the peripheral blood; an enzyme‐linked immunosorbent assay (ELISA) for determining the absolute and relative quantities of various Hb species and one‐step immunolabeling for fluorescence microscopic and flow cytometric analyses of the distribution of RBC with respect to their Hb types. We utilized these methods for monitoring the Hb F level and the percentage of Hb F‐containing cells in patients with sickle cell anemia undergoing treatment with hydroxyurea.