重组人3型腺病毒六邻体蛋白纯化及抗原性检测  

Purification and antigenicity detection of recombinant hexon of human type-3 adenovirus

作　　者：郑红霞[1] 刘勇[1] 曲章义[1] 郭彩玲[1] 王立群[1] 崔洪波[1] 

机构地区：[1]哈尔滨医科大学微生物学教研室,黑龙江哈尔滨150086

出　　处：《哈尔滨医科大学学报》2005年第6期471-473,478,共4页Journal of Harbin Medical University

基　　金：中国医学科学院青年基金资助项目(9331014)

摘　　要：目的纯化重组人3型腺病毒六邻体蛋白并检测其抗原性。方法在原核表达系统中高效表达重组人3型腺病毒六邻体蛋白。利用Ni-NTA琼脂糖亲和层析柱进行蛋白纯化。用纯化的重组六邻体蛋白免疫新西兰家兔,利用免疫印迹技术检测其血清中抗体效价以鉴定该蛋白的免疫原性和反应原性。结果得到纯化的重组人3型腺病毒六邻体蛋白,该蛋白免疫后产生高效价抗体,该抗体能与重组六邻体蛋白、含有该蛋白的重组工程菌株以及人3型腺病毒发生特异性的免疫印迹反应,证明该重组六邻体蛋白具有良好的抗原性和3型腺病毒特异性。结论所获得的重组人3型腺病毒六邻体蛋白具有良好的抗原性和特异性,为开发基因工程疫苗奠定了基础。Objective To purify the recombinant hexon of human type-3 adenovirus and detect its antigenicity. Methods Under the optimum expression conditions, the hexon protein of human type 3 adenovirus was efficiently expressed in E. coll. The expressed proteins were purified by means of Ni-NTA affinity chromatography. Using the purified hexon to immune rabbit and test the hexon＇ s antigenicity with the method of Westernblot technique. Results The hexon of human type 3 adenovirus was purified and was proved to immune the rabbit with high antibody titer and the antibody reacted specific with the purified recombinant hexon, the recombinant E. coli strain containing the hexon protein and the type 3 adenovirus itself. Thus it was proved that the recombinant protein possessed both of the high antigenicity and high specificity to type 3 adenovirus. Conclusion The recombinant hexon has high antigenicity and specificity.

关 键 词：腺病毒 六邻体 蛋白纯化 抗原性 

分 类 号：R373.1[医药卫生—病原生物学]