Simple and rapid enzyme-linked immunosorbent assay for the detection of hemoglobin C[alpha 2 beta 2 6(A3)Glu----Lys] in cord blood using a monoclonal antibody.
We have generated a murine hybridoma that secretes a monoclonal antibody (mAb) that is highly specific for hemoglobin C (HbC) [alpha 2 beta 2 6(A3)Glu----Lys] and shows no cross reactivity with HbA, HbA2, HbF, 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 microliters/well of whole blood or 5 micrograms 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 beta 6GLU of HbA, allows the discrimination between HbC homozygotes, heterozygotes, and normals.
Department of Cell and Molecular Biology, Medical College of Georgia, Augusta 30912-2100.
M
M
Shyamala
C R
CR
Kiefer
H
H
Moscoso
F A
FA
Garver
D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005312:Fetal Blood; D006444:Hemoglobin C / Q000032:analysis* / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D006444:Hemoglobin C