Table 1.

Specificity of the 384 selected antibody clones obtainted from the second round of selection against ε-Hb, tested in ELISA towards different antigens. E: recombinant denatured ε-Hb. RBC: parental red blood cell extract (containing α- and β-Hb, but no ε-Hb), BL21: bacterial control cell extract, and K562: K562 cell extract (containing α-, γ-, δ-, ε-, and ζ-Hb). Neg.: amount of clones not recognising any of the tested antigens. E+RBC, E+BL21, E+K562, and E+K562−BL21−RBC, amount of clones recognising ε-Hb (E) together with other antigens. A total of 178 (46%) of the tested clones (384) gave positive ELISA signal toward ε-Hb, while 57 (15%) recognised K562 cell extract. Only two clones recognised K562 cell extract without recognising ε-Hb. Twelve of the selected antibodies showed specificity of the ε-Hb, since they recognised the denatured recombinant ε-Hb, the K562 extract, containing native ε-Hb and not the parental RBC, nor the BL21 extract.

E	RBC	BL21	K562	E+RBC	E+BL21	E+K562	E+K562−BL21−RBC	Neg.
178	10	30	57	10	19	55	12	206