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Simple and rapid enzyme‐linked immunosorbent assay for the detection of hemoglobin C[α 2 β 2 6(A3)Glu → Lys] in cord blood using a monoclonal antibody
Author(s) -
Shyamala M.,
Kiefer Charles R.,
Moscoso Hugo,
Garver Fred A.
Publication year - 1990
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/ajh.2830330308
Subject(s) - cord blood , monoclonal antibody , serial dilution , antibody , microbiology and biotechnology , hemoglobin , monoclonal , whole blood , antigen , chemistry , immunology , medicine , biology , biochemistry , pathology , alternative medicine
We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [α 2 ß 2 6(A3)Glu→Lys] and shows no cross reactivity with HbA 2 , HbF 2 , HbS, HbE, or Hb O‐Arab. Using this antibody, we developed a simple and rapid enzyme linked immunosorbent assay (ELISA) technique for the detection of HbC in both adult and cord blood. The assay can be carried out using either whole blood samples or hemolysates. With as little as 10 μl/well of whole blood or 5 μg Hb/well of hemolysates, and, with dilutions of the antibody up to 10 −5 we were able to detect HbC unequivocally in cord blood samples. The ELISA procedure could detect HbC in proportions as low as 0.01%. This simple diagnostic test represents a technological advance in Hb identification and can easily be used for mass screening (96 samples in less than 45 min) to detect HbC. Furthermore, this assay, when employed in conjunction with an mAb specific for β6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals.