[Preparation and identification of monoclonal antibodies against different epitopes on human zeta globin chain].

OBJECTIVE

To prepare and identify the monoclonal antibodies (mAb) against human zeta globin chain.

METHODS

BALB/c mice were immunized with purified recombinant zeta globin chain, and the hybridomas were generated by fusion of mouse spleen cells and myeloma cells NS-1. After three fusions and successive cloning, 3 hybridoma cell lines secreting the mAb against zeta were obtained and the antibodies were purified from the ascites, followed by identification with indirect enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA in combination with rabbit anti-zeta serum.

RESULTS AND CONCLUSION

Three hybridoma cell lines secreting anti-zeta mAb were established, designated as 1A12, 3H9 and 4D11, respectively. Both of 3H9 and 4D11 mAbs belonged to IgG1 isotype and mAb 1A12 to IgG2a. All the mAbs could bind specifically to recombinant zeta and natural zeta globin from hemolysate of --(SEA) gene carrier. In addition, 1A12 and 3H9 mAbs could recognize different epitopes on zeta globin, suggesting the possibility of developing a detection system for screening alpha-thalassemia with these mAbs.