卵巢癌抗独特型抗体6B11scFv和人粒细胞巨噬细胞集落刺激因子融合蛋白的构建和表达  

The Construction and Expression of 6B11scFv Antiidiotypic Antibody and Human Granulocytemacrophage colony Stimulating Factor in Escherichia Coli

作　　者：杨飞雪[1,2] 钱和年[1,2] 黄华梁[1,2] 冯捷[1,2] 崔恒[1,2] 

机构地区：[1]北京医科大学人民医院妇科肿瘤中心 [2]中国科学院遗传所

出　　处：《中华妇产科杂志》1999年第7期417-419,共3页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金

摘　　要：目的构建和表达模拟人卵巢癌抗原的抗独特型单链抗体６Ｂ１１ｓｃＦｖ和人粒细胞巨噬细胞集落刺激因子（ｈＧＭＣＳＦ）的融合蛋白，以提高抗独特型抗体的免疫原性，为进一步用于卵巢癌的免疫治疗提供基础。方法采用基因工程技术，用编码连接肽的ＤＮＡ连接６Ｂ１１ｓｃＦｖ和ｈＧＭＣＳＦｃＤＮＡ，并对大肠杆菌用温度进行诱导表达；采用蛋白免疫吸附印迹法，对表达的蛋白进行活性测定。结果表达的蛋白能分别与６Ｂ１１的初始抗体和小鼠抗ｈＧＭＣＳＦ单克隆抗体特异结合。结论表达的融合蛋白保留了６Ｂ１１ｓｃＦｖ和ｈＧＭＣＳＦ两种成分的活性，增强了６Ｂ１１ｓｃＦｖ的免疫原性。Objective To enhance the antigenicity of an antiidiotypic single chain antibody 6B11 mimicking ovarian cancer antigen. Methods Using DNA recombinant technique, a recombinant fusion protein expression vector was constructed. This vector linked 6B11scFv cDNA and human GMCSF with a linker. Protein productions were induced with the temperature change in E. Coli and analyzed with western blot. Results The expressed fusion protein can be reacted with both antihuman antibody and COC1669(AB1 of 6B11). Conclusion The recombinant fusion protein keeps the activities of both components and may be used as tumor vaccine for ovarian cancer.

关 键 词：重组融合蛋白质 单链抗体 HGM-CSF 卵巢肿瘤 

分 类 号：R737.310.2[医药卫生—肿瘤]