[Preparation of high-affinity monclonal antibody against hemoglobin delta globin chain and beta globin chain].

Aim: To prepare monclonal antibody (mAb) against both hemoglobin A2 (HbA2) and hemglobin A (HbA), this antiboy can bind to delta globin chain and beta globin chain, but not to gamma globin chain.
Methods: Mice was immunized with recombinant Hb delta fusion protein, and hybridoma cells were generated by cell fusion techniques followed by screening with natural HbA2 and HbA separated by ion exchange chromatography. The purified monclonal antibody was identified by indirect ELISA, Western blot based on native PAGE and SDS-PAGE, surface plasmon resonance (SPR), flow cytometry and immunohistochemistry.
Results: The monoclonal antibody against both HbA2 and HbA was obtained and designated as 2C9 that shows no binding to fetal hemoglobin (HbF), alpha globin chain and recombinant zetaglobin chain.
Conclusion: The mAb 2C9 was defined as specificity to hemoglobin delta globin chain and beta globin chain, which suggests that mAb 2C9 recognizes a common epitope on bothdeltaglobin chain and beta globin chain. This antibody would be expected to be an effective tool in research and clinical practice in hemoglobinopathies.