Use of specific fluorescent antibodies for the identification of hemoglobin C in erythrocytes

Abstract

Antibodies against hemoglobulin C (alpha2beta2 6Glu leads to Lys) were produced by immunizing horses and were purified by affinity chromatography. As expected from the bivalency of both the antibody and the antigen, the purified antibodies failed to produce immunoprecipitates upon reaction with the corresponding antigens. Identification of hemoglobin C in individual erythrocytes was achieved by reacting the fluorescein isothiocyanate-conjugated antibodies with the hemoglobin antigen in fixed smears of peripheral blood. Red cells from persons having a hemoglobin C gene were labeled strongly upon reaction with anti-Hb C-FITC; there was no labeling of red cells containing normal hemoglobins or Hb S, suggesting that the anti-Hb C antibodies recognize only the amino-terminal segment of the beta chains that contain lysine in position beta6.