HPV16主要衣壳蛋白L1在昆虫细胞中的表达及免疫学活性分析

Expression of HPV16 L1 protein in insect cells and analysis of its immunoactivity

目的 在昆虫细胞中表达HPV16L1(m2 0 2 )蛋白 ,并分析其免疫学活性。方法 构建表达载体pFASTBACHTb L1(m2 0 2 ) ,用重组病毒感染sf9细胞表达HPV16L1(m2 0 2 )蛋白 ,SDS PAGE、western blot鉴定其表达 ,亲和层析和离子交换层析纯化后的产物经鼻腔免疫Balb/c小鼠 ,竞争抑制ELISA分析免疫血清的中和活性。结果 SDS PAGE、western blot结果证明HPV16L1(m2 0 2 )蛋白的表达 ,纯化复性后产率约为 17% ,免疫血清具有竞争抑制HPV中和单抗与HPV16L1(m2 0 2 )相结合的能力。结论 昆虫细胞表达的HPV16L1(m2 0 2 )

Objective To express and analyse the immune activity of HPV16 L1(m202) protein in insect cells sf9.Methods Expression vector pFASTBAC HTb L1(m202)had been constructed ,sf9 cells were transfected with recombinant viruses and expressed HPV16 L1(m202) protein. The products confirmed by SDS PAGE, western blot, purified by affinity chromatography and ion exchange chromatography, were nasal immunized Balb/c mice, the neutralizing activity of anti serum was analysed by ELISA.Results SDS PAGE and Western blot showed that HPV16 L1(m202) protein had been expressed in sf9 cells, the efficiency of purification and renaturation was approximately 17%, the anti serum has the ability of competing inhibition HPV16 McAb reacting with HPV16 L1(m202) protein.Conclusion HPV16 L1(m202) protein expressed in insect cells has the potentiality of being prophylactic vaccine.