Enzyme Immunoassay for the Identification of Hemoglobin Variants

Abstract

We have prepared monospecific antibodies to Hbs D-Los Angeles, J-Blaltimore, O-Arab and J-Paris-I and developed an enzyme immunoassay (ELISA) for their identification in hemoly-sates. Hbs in adult or cord blood hemolysates were coated to the wells of microtiter plates and reacted with the appropriate antisera followed by the detection system which contains anti-rabbit IgG/peroxidase conjugate and the substrate tetramethyl-benzidine. Sixty-nine samples were tentatively considered to contain the above hemoglobin variants by isoeleotrofocusing and the identity of 83% of them was confirmed by ELISA Some of the non-reacting hemolysates were shown by amino acid sequence analysis to contain Hbs Korle-Bu, D-Ibadan, G-Copenhagen and the new variant Chandigarh. This ELISA offers specificity and simplicity for the confirmatory identification of hemoglobin variants.