Xu Wan Xiang, He Ya Ping, Qiu De Yi, Liao Mao Chuan, Hong Ai Zhen, Ji Zhao Neng, Gu Shao Hua, Chen Jin Zhong, Xie Yi
Department of Reproductive Biology, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2006 Oct;39(5):414-22.
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 beta subunit (beta-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 beta-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 12 L culture. Also, the CPl and CP10 immunogens can induce antibodies in mice that recognize recombinant CP1 betar CP10 and natural beta-hCG, and there are three anti-beta5, beta9 and beta8 BCE antibodies in their antisera. The construction and expression of beta-hCG CP1 and CP10 will provide new immunogens for developing an ideal and superior hCG birth control and/or tumor therapeutic vaccine.
已设计出两种生物合成嵌合肽(CP)免疫原,分别命名为CP1和CP10,它们由人绒毛膜促性腺激素β亚基(β-hCG)的三个线性B细胞表位(BCE)和六个外源T细胞表位组成,其中包括来自乙肝表面抗原和破伤风类毒素的两个“通用”T细胞表位。合成了编码CP1及其衍生物CP10的两个人工基因,当它们分别插入热诱导载体后,可在大肠杆菌中表达,表达水平约占细胞总蛋白的1%。在蛋白质印迹试验中,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(PAGE)凝胶上显示的约16.5 kD的表达CP1和CP10蛋白可被针对β-hCG每个线性表位的单克隆或多克隆抗体识别。使用我们改进的制备性凝胶PAGE方法,每种表达蛋白均可纯化至相对纯度为95%。每12 L培养物的产量约为1 - 2 mg。此外,CP1和CP10免疫原可在小鼠体内诱导产生能识别重组CP1和CPb10以及天然β-hCG的抗体,其抗血清中存在三种抗β5、β9和β8 BCE抗体。β-hCG CP1和CP10的构建与表达将为开发理想且优质的hCG避孕和/或肿瘤治疗疫苗提供新的免疫原。