Development of a screening system for detection of somatic mutations. I. Enzyme immunoassay for detection of antibodies against specific hemoglobin determinants.
A solid-phase enzyme immunoassay for the detection of antibodies, specific for hemoglobin (Hb) is described. The application of glutaraldehyde resulted in a sensitive assay and allowed the use of urea, which is an important advantage if polypeptides not soluble in aqueous buffers are to be used. Mutation-carrying Hb chains can be purified, solubilized in urea and used in the immunoassay to monitor the purification and selection of antibodies specific for these variants. Specific antibodies are the main tools for the development of a hemoglobin-locus mutation system for detection of potentially mutagenic environmental agents. With erythrocytes as target cells, this system permits in vivo monitoring of subjects under exposure. Conventional antibody production, however, frequently turns out to be unsuccessful. The production of monoclonal antibodies has several advantages over conventional antibody production, but a sensitive antibody screening system is essential. Because of the sensitivity and the ease with which a large panel of antibody fractions against a vast panel of Hb antigens can be examined, the described immunoassay has potential value for the screening of hybridoma cultures.
D000328:Adult; D000818:Animals; D001202:Ascitic Fluid / Q000032:analysis / Q000276:immunology; D001666:Binding Sites, Antibody; D000939:Epitopes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D005976:Glutaral / Q000494:pharmacology; D006454:Hemoglobins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007124:Immunoenzyme Techniques; D007518:Isoantibodies / Q000032:analysis*; D051379:Mice; D009154:Mutation; D011309:Preservation, Biological; D011817:Rabbits; D014508:Urea / Q000494:pharmacology
D000939:Epitopes; D006454:Hemoglobins; D007106:Immune Sera; D007518:Isoantibodies; D014508:Urea; D005976:Glutaral