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Application of a monoclonal antibody specific for the δ chain of hemoglobin A2 in the diagnosis of β thalassemia

We have developed a murine monoclonal antibody (mAb) specific for the δ chain of hemoglobin (Hb) A2 that does not cross‐react with α, β, or γ chains. The mAb reacted with Hb P‐Nilotic (β δ hybrid), but not with Hb Lepore‐Boston (δ β hybrid), indicating an epitope consisting of positions 116 (Arg) and 117 (Asn) or 125 (Gin) and 126 (Met) of the δ chain. By using this antibody, we have established a simple and rapid enzyme‐linked immunosorbent assay (ELISA) for the detection and quantification of Hb A2 in adult, cord, and fetal hemolysates. We analyzed 70 adult, 8 newborn, and 19 fetal hemolysates from normal subjects and those with various hemoglobinopathies. The mean percentage of Hb A2 was 2.5 for normal adults, 5.4 for β thalassemic (β thai) heterozygotes, and <0.1% in β thal fetal samples. We were able to distinguish and characterize certain phenotypes of β thai patients such as β thai heterozygotes, β° thai homozygotes, and Cβ° thal, and Cβ° thal double heterozygotes with the aid of this and other mAbs we have generated. This technique is a valuable addition to current methods for the diagnosis of β thai based on quantification of Hb A2.