多表位hCG嵌合肽(CP1和CP10)抗原的构建及其免疫原性  

CONSTRUCTION AND IMMUNOGENICITY OF hCG CHIMERIC PEPTIDES (CP1 and CP10) CONTAINING THREE SELF-B CELL EPITOPES AND SIX FOREIGN T CELL EPITOPES

作　　者：徐万祥[1] 何亚萍[1] 邱德义[1] 廖矛川[1] 洪爱真[1] 季朝能[2] 顾少华[2] 陈金中[2] 谢毅[2] 

机构地区：[1]上海市计划生育科学研究所生殖生物学研究室 [2]复旦大学生命科学院遗传所遗传工程国家重点实验室,上海200433

出　　处：《分子细胞生物学报》2006年第5期414-422,共9页Journal of Molecular Cell Biology

基　　金：国家自然科学基金(No.39870856和No.30070792)资助。~~

摘　　要：本研究设计了CP1和CP10两种嵌合肽免疫原,它们由人绒毛膜促性腺激素β亚单位(β- hCG)3个线性B细胞表位(BCE)以及包括来自乙肝表面抗原和破伤风类毒素2个广谱性T细胞表位(TCE)的6个TCEs组合而成。编码CP1及其衍生物CP10的人工基因分别重组插入热诱导pBV221载体后,均能在大肠杆菌中以约占细菌总蛋白1％的比例被表达。在蛋白印迹试验中,在SDS-聚丙烯酰胺凝胶电泳凝胶上显示约为16．5 kD分子量的CP1和CP10表达蛋白,能被抗各插入表位单克隆或多克隆抗体识别。各表达蛋白可用制备性PAGE方法一步纯化,纯化产物均显示95％以上的电泳均一性,纯化得率可达到每升培养液1-2mg水平。以它们为抗原主动免疫小鼠,都能分别产生识别CP1或CP10和天然β-hCG的抗血清,并都能在它们的抗血清中检测到各插入抗原表位的抗体。hCG CPs的可获得性及其能诱导各表位抗体生成特征,有可能成为研制hCG抗生育和肿瘤治疗性疫苗的新的候选免疫原。Two bio-synthesizing chimeric peptide （CP） immunogens named CP1 and CP10 have been designed,which consist of three linear B cell epitopes （BCE） of human chorionic gonadotropin β subunit （β-hCG） and six foreign T cell epitopes including two ＂promiscuous＂ TCEs from hepatitis B surface antigen and tetanus toxoid. Two artificial genes encoding CP1 and its derivative CP10 were synthesized,which could be expressed in E.coli at the level of about 1% of the total cell proteins when inserted into the thermo-induction vector respectively. In Western blot tests,the expressed CP1 and CP10 proteins with about 16.5 kD shown on the SDS-polyacrylamide gel electrophoresis （PAGE） gel can be recognized by the monoclonal or polyclonal antibodies specific to each linear epitope of β-hCG. Each of expressed proteins can be purified with 95% relative homogeneity using our improved method of preparative gel PAGE. Their yields were about 1- 2 mg per 1 L culture. Also,the CP1 and CP10 immunogens can induce antibodies in mice that recognize recombinant CP1 or CP10 and natural β-hCG,and there are three anti-β5,β9 and β8 BCE antibodies in their antisera. The construction and expression of β-hCG CP1 and CP10 will provide new immunogens for developing an ideal and superior hCG birth control and/or tumor therapeutic vaccine.

关 键 词：人毛膜促性腺激素β亚单位嵌合肽免疫原 原核表达 纯化 免疫应答 

分 类 号：R392[医药卫生—免疫学] Q789[医药卫生—基础医学]