Expression and purification of nucleocapsid protein of hantaan virus and creation of anti-NP monoclonal antibody

  Objective   To prepare stable and unique recombined nucleocapsid protein(rNP)and monoclonal antibody (McAb)of Hantaan virus(HTNV).

  Method   To establish prokaryotic expression carriers of HTNV strain 76118 which are pBV2202S113 and pET28a2S113 through inducing in E.coli.The bacterial products were analyzed by using SDS2PA GE and Western-Blot.The results of analysis showed that there was a special antigen about 48kD, whick was rNP of HTNV.Myeloma cell SP2/0 were hybridized with lymphocytes B which taken from Immunized Balb/C.The positive hybridoma were screened with rNP antigen derived from pET28a2S113 purified by DEAE-Sephadex A50 and Ni-NAT chromatography.To establish cell line and select two lines for producing McAb of rNP routinely.Finally, McAb was identified.

  Results   Rceombination prokaryotic expression carriers pBV2202S113 and pET28a2S113 were constructed for producing nucleoprotein of HTNV successfully.High purity rNP and high titer Mc/Ab of anti-NP which were 2 E6 and 4D8 were obtained.

  Conclusion   The rNP and McAb of HTNV have great value in epidemiological sdurveillance, diagnosis and scientific research of epidemic hemorrhagic fever.