Enzyme immunoassay for the identification of hemoglobin variants

We have prepared monospecific antibodies to Hbs D-Los Angeles, J-Baltimore, O-Arab and J-Paris-I and developed an enzyme immunoassay (ELISA) for their identification in hemolysates. Hbs in adult or cord blood hemolysates were coated to the wells of microtiter plates and reacted with the appropriate...

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Bibliographic Details
Published inHemoglobin Vol. 14; no. 4; p. 389
Main Authors Moscoso, H, Kiefer, C R, Shyamala, M, Garver, F A
Format Journal Article
LanguageEnglish
Published England 1990
Subjects
Adult
Animals
Antibody Specificity
Enzyme-Linked Immunosorbent Assay
Hemoglobin J - analysis
Hemoglobin J - immunology
Hemoglobins, Abnormal - analysis
Hemoglobins, Abnormal - immunology
Humans
Immune Sera
Immunoenzyme Techniques
Infant, Newborn
Isoelectric Focusing
Rabbits
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Summary:We have prepared monospecific antibodies to Hbs D-Los Angeles, J-Baltimore, O-Arab and J-Paris-I and developed an enzyme immunoassay (ELISA) for their identification in hemolysates. 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 tetramethylbenzidine. Sixty-nine samples were tentatively considered to contain the above hemoglobin variants by isoelectrofocusing 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.
ISSN:0363-0269
DOI:10.3109/03630269009031999