[Preparation and characterization of the epitopes of SARS coronavirus spike protein].

Fang, Kai; Zhong, Lian-Sheng; Ma, Ru-Hai; Wang, Shao-Cheng; Pan, Zhong-Cheng; Zhang, Yu-Kui; He, Qun; Zhao, Yu-Jie

AIM To prepare the recombinant epitopes of SARS-CoV spike protein and study their antigenic property to spike protein. METHODS The epitopes of SARS-CoV spike protein were analyzed by epitope prediction software. A novel gene, named Z, encoding 16 predicted epitopes of spike protein was designed and synthesized using chemical method. Z gene was cloned into vector pET-28a(+), expressed in E.coli BL21(DE3) and purified by Ni(2+) affinity method. Z protein was used as antigen to immunize the rabbit and anti-Z sera was collected. Then the antigenic property of Z protein to SARS-CoV spike protein was analyzed by dot-blot and ELISA assay. RESULTS Z gene was successfully designed and expressed in E.coli BL21(DE3). Dot blot analysis showed that SARS-CoV spike protein, which was expressed and purified from mammal cells, can be detected by anti-Z sera from rabbit. ELISA analysis indicated the SARS-CoV antigen prepared from SARS-CoV lysates can be detected by anti-Z sera. CONCLUSION The predicted epitopes of Z protein can induce the development of SARS-CoV spike protein antibody in rabbits, which provides a new protein for developing vaccine against SARS-CoV.