Preparation and identification of recombinant human hemoglobin δ chain.

Objective To prepare recombinant human hemoglobin δ chain for the establishment of HbA2 immunoassay in the screening of β thalassemia.Methods Hbδ gene was cloned from K562 cells and amplified by RT-PCR.Two expression vectors were constructed.The fusion proteins were expressed in E.coli and identified by Western blotting and ELISA.Results Two expression vectors,namely pET32-Hbδ and pET43.1-Hbδ,were constructed and expressed in inclusion and soluble phase.The fusion protein was purified and was confirmed by Western blot and ELISA as Hbδ.Conclusion Two recombinant human hemoglobin δ chains were obtained.The recombinant proteins may be used as immunogens for immunoassay of HbA2 in the screening of βthalassemia.