Monoclonal antibody-based methods for quantitation of hemoglobins: application to evaluating patients with sickle cell anemia treated with hydroxyurea.
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.
Epstein
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Isolab Inc., Akron OH 44321, USA.
Epstein
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M
Boulet
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A
Fibach
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E
Rodgers
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GP
D000755:Anemia, Sickle Cell / Q000097:blood / Q000188:drug therapy*; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D005319:Fetal Hemoglobin / Q000032:analysis; D005434:Flow Cytometry; D006451:Hemoglobin, Sickle / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D006918:Hydroxyurea / Q000627:therapeutic use*; D007118:Immunoassay / Q000379:methods*; D008856:Microscopy, Fluorescence
D000911:Antibodies, Monoclonal; D006451:Hemoglobin, Sickle; D006454:Hemoglobins; D005319:Fetal Hemoglobin; D006918:Hydroxyurea