梅毒螺旋体多表位融合抗原的表达及免疫性研究

Expression and Immunological Characteristics of Multiepitope Fusion Antigen of Treponema Pallidum

设计梅毒螺旋体多表位融合抗原,该融合抗原含外膜蛋白Tp0453,Tp0435,Tp1038,Tp0868和Tp0965重要抗原表位,人工合成该多表位融合抗原基因,转入大肠杆菌中IPTG诱导表达,镍柱亲合层析纯化.利用多表位融合抗原检测人血清中梅毒特异性抗体,梅毒患者样本组与非患者样本组检测结果存在极其显著性差异,说明多表位融合抗原可被梅毒抗体特异性识别,有较好免疫反应性,可作为梅毒诊断试剂新抗原.利用多表位融合抗原免疫新西兰兔,兔血清中可持续检测到高滴度梅毒抗体,显示该多表位融合抗原有较强免疫原性,是梅毒基因工程疫苗候选蛋白.

In this work,the multiepitope fusion antigen gene was designed and synthesized,which contained the important epitopes of Tp0453,Tp0435,Tp1038,Tp0868 and Tp0965 from Treponema pallidum.The multiepitope fusion antigen was expressed in Escherichia coli with IPTG and purified by Nickel-affinity chromatography column.Then,the fusion antigen was used to analyze the human serum antibody of a syphilis patient group and a non-syphilis patient group,and a statistically significant difference for the syphilis antibody was observed between the two groups.ELISA results showed that the fusion antigen could be recognized by syphilis antibody with specificity.It had good reactinogenicity,and thus may be a good antigen for syphilis detection.A rabbit was injected with the fusion antigen and we found that it could produce syphilis antibody effectively.Our present results suggest that the multiepitope fusion antigen possesses good immunogenicity and should be an ideal candidate for the syphilis vaccine.