Preparation and identification of monoclonal antibodies against different epitopes on human ζ globin chain

Abstract

Abstract: Objective To prepare and identify the monoclonal antibodies (mAb) against human ζ globin chain. Methods BALB/c mice were immunized with purified recombinant ζ 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 ζ 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-ζ serum. Results and Conclusion Three hybridoma cell lines secreting anti-ζ 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 ζ and natural ζ globin from hemolysate of --^SEA gene carrier. In addition, 1A12 and 3H9 mAbs could recognize different epitopes on ζ globin, suggesting the possibility of developing a detection system for screening alpha-thalassemia with these mAbs.

CLC Number:

R392.11

TANG Lei¹, ZHU Ping¹, LUO Chen², XU Xiang-ming², FU Ning¹. Preparation and identification of monoclonal antibodies against different epitopes on human ζ globin chain[J]. Journal of Southern Medical University, 2005, 25(11): 1394-1397.

References

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