Abstract: Objective To prepare a specific antibody against human hemoglobin (Hb) by hybridoma technique, which has great significance in further exploring the functions of Hb in early diagnosis and pathogenesis of Parkinson's disease.Methods The fusion cells were produced with spleen cells of two Balb/c mice immunized with recombinant human Hb to combine with homologous myeloma cells. A positive clone, 5B1, was selected by screening the hybridoma cells with the enzyme-linked immunosorbent assay (ELISA) after three times of limited dilution method. The antibody was acquired by intraperitoneal injection in immunosuppressed BALB/c mice. And Dot blotting, ELISA, Co-immunoprecipitation (Co-IP),Western blotting tests and immunohistochemical (IHC) staining were used for further testing. Results One stable hybridoma cell line, named as 5B1, was found to secret anti-Hb with high titers and specificity. Conclusion The results demonstrated that the monoclonal antibody made in this study has specificity to Hb in human blood, monkey, rats and mouse, and offers a powerful tool for future studies on Hb in Parkinson's disease (PD).

Key words: hemoglobin (Hb), monoclonal antibody, Parkinson's disease