Abstract: AIM: To express and purify HCV core binding protein ( HCBP12) with prokaryotic cell ex-pressive vector and to prepare HCBP12 specific rabbit polyclonal antibody .METHODS: The DNA segment of HCBP12 was amplified by RT-PCR with mRNA tem-plate from HepG2 cells and inserted into inducible pro-karyotic expressive vector pET-32a(+) .The recombi-nant plasmid, pET-32a(+)-HCBP12, was trans-formed into the competent E.coli BL21 .The inducible HCBP12 fusion protein was analyzed by SDS-PAGE and conformed by Western blot .Then the inducible HCBP12 protein w as purified and renatured by Ni⁺ affirity column chromatography. The purified HCBP12 ploteinwas used to immunize New Zea land rabbits for preparing polyclonal antibody. The specificity and potency of polyclonal antibody was evaluated by Western blot and ELISA .RESULTS: The HCBP12 fusion pro-tein was highly expressed and purified .The polyclonal antibody against HCBP12 was obtained successfully with the titer >1:512000 and confirmed specifically with Western blot .CONCLUSION: The successful ex-pression and purification of HCBP12 fusion protein and the preparation of specific anti-HCBP12 polyclonal an-tibody will be valuable for the study on the biological function of HCBP12 and clinical therapy of Hepatitis C .

Key words: HCBP12, fusion protein, protein pu-rification, polyclonal antibody