Development of a somatic mutation screening system using Hb mutants. IV. Successful detection of red cells containing the human frameshift mutants Hb Wayne and Hb Cranston using monospecific fluorescent antibodies.
The production and purification of antibodies detecting Hb Wayne, an alpha-globin frameshift mutant, and Hb Cranston, a beta-globin frameshift mutant, are described. The antibodies are of a nonprecipitating nature, and they permit strong fluorescent labeling of erythrocytes containing Hb Wayne or Hb Cranston. Studies using artificial mixtures containing cells with either of the two mutants in frequencies ranging from 1 in 10(2) to 1 in 10(5) showed that fluorescent antibodies can detect rare mutant red cells in the presence of vast excesses of normal erythrocytes. On the basis of the structures and the molecular lesions underlying production of the two abnormal hemoglobins, we predict that the anti-Hb Wayne antibody will detect several frameshift mutants resulting from deletion of 3n + 1 nucleotides or insertion of 3n + 2 nucleotides at the 5' side of the codon normally specifying residue 139 of the alpha chain. The anti-Hb Cranston antibody should be capable of detecting beta chains, the corresponding genes of which have sustained insertions of 3n + 2 nucleotides or deletions of 3n + 1 nucleotides on the 5' side of the codon normally specifying residue 144. The two antibodies may, therefore, prove to be valuable in the development of a system aimed at detecting rare erythrocytes that express mutations which arise in the hemopoietic stem cells of normal individuals and subjects exposed to mutagens.
D000906:Antibodies; D004912:Erythrocytes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005820:Genetic Testing / Q000379:methods; D006455:Hemoglobins, Abnormal / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D006801:Humans; D009153:Mutagens; D009154:Mutation
D000906:Antibodies; D006455:Hemoglobins, Abnormal; D009153:Mutagens