摘要
AIM: To analyze the amino acid sequences of hypervariable region 1 (HVR1) of HCV isolates in China and to construct a combinatorial chimeric HVR1 protein having a very broad high cross-reactivity.METHODS: All of the published HVR1 sequences from China were collected and processed with a computer program.Several representative HVR1's sequences were formulated based on a consensus profile and homology within certain subdivision. A few reported HVR1 mimotope sequences were also included for a broader representation. All of them were cloned and expressed in E.coli. The cross-reactivity of the purified recombinant HVR1 antigens was tested by ELISA with a panel of sera from HCV infected patients in China.Some of them were further ligated together to form a combinatorial HVR1 chimera.RESULTS: Altogether 12 HVR1s were selected and expressed in E. coli and purified to homogeneity. All of these purified antigens showed some cross-reactivity with sera in a 27 HCV positive panel. Recombinant HVR1s of No. 1, 2, 4, and 8# showing broad cross-reactivities and complementarity with each other, were selected for the ligation elements. The chimera containing these 4 HVR1s was highly expressed in E. coli. The purified chimeric antigen could react not only with all the HCV antibody positive sera in the panel but also with 90/91 sera of HCV -infected patients.CONCLUSION: The chimeric antigen was shown to have a broad cross-reactivity. It may be helpful for solving the problem caused by high variability of HCV, and in the efforts for a novel vaccine against the virus.
AIM:To analyze the amino acid sequences of hypervariable region 1 (HVR1) of HCV isolates in China and to construct a combinatorial chimeric HVR1 protein having a very broad high cross-reactivity. METHODS:All of the published HVR1 sequences from China were collected and processed with a computer program. Several representative HVRI's sequences were formulated based on a consensus profile and homology within certain subdivision.A few reported HVR1 mimotope sequences were also included for a broader representation.All of them were cloned and expressed in E.coli.The cross-reactivity of the purified recombinant HVR1 antigens was tested by ELISA with a panel of sera from HCV infected patients in China. Some of them were further ligated together to form a combinatorial HVR1 chimera. RESULTS:Altogether 12 HVR1_s were selected and expressed in E.coliand purified to homogeneity.All of these purified antigens showed some cross-reactivity with sera in a 27 HCV positive panel.Recombinant HVRls of No.1,2,4,and 8# showing broad cross-reactivities and complementarity with each other,were selected for the ligation elements.The chimera containing these 4 HVRls was highly expressed in E.coli.The purified chimeric antigen could react not only with all the HCV antibody positive sera in the panel but also with 90/91 sera of HCV -infected patients. CONCLUSION:The chimeric antigen was shown to have a broad cross-reactivity.It may be helpful for solving the problem caused by high variability of HCV,and in the efforts for a novel vaccine against the virus.
基金
National 10~(th)Five-Year Plan of Science and Technology Brainstorm Project,No.2001BA708B06
a grant from Natural Science Foundation of Beijing,No.7002031
