基于抗原表位的抗蛋白质抗体制备新方法  

A novel method to prepare the anti-protein antibodies based on antigen epitope

作　　者：周游[1] 吴峰[1] 范国才[1] 吴祖泽[1] 王清明[1] 

机构地区：[1]蛋白质组学国家重点实验室,北京蛋白质组研究中心,军事医学科学院放射与辐射医学研究所,北京102206

出　　处：《现代免疫学》2009年第5期364-369,共6页Current Immunology

基　　金：国家重大科学研究计划资助项目(2006CB910803)

摘　　要：为建立一种基于抗原表位的抗蛋白质抗体制备新方法。以T7噬菌体作为展示载体,在其表面展示蛋白质的抗原表位肽,提取重组噬菌体作为免疫原免疫动物,通过ELISA、Western blot和细胞免疫荧光技术检测抗体的滴度及特异性。结果表明,以展示蛋白质抗原表位的T7噬菌体免疫动物,得到的抗体能特异识别相应的表位肽,其中针对Pif1-α抗原表位的抗体,还能特异识别Pif1-α蛋白。结论是,表面展示小肽的T7噬菌体具有很好的免疫原性,如果展示的小肽确为蛋白质抗原表位,则可作为完整蛋白质抗原的替代抗原免疫动物来制备识别完整蛋白的抗体。To find out a novel method to prepare the anti-protein antibodies based on antigen epitope, epitope peptide was displayed on T7 bacteriophage, and the recombinant T7 bacteriophage was extracted and used to immunize animals. The antibody titer and specificity were analyzed by ELISA, Western blotting and immunofluorescence. The antibody derived from animals immunized by epitope peptide displayed on T7 phages could specifically recognize the epitope peptide. One of them, the antibody against the epitope peptide of Pifl-α, could also specifically recognize Pifl-α protein. It is evident that T7 phage displayed peptides possessed good immunogenicity. If this small displayed peptide is a real antigenic epitope of the protein antigen, it can be used as an alternative antigen to immunize animals to prepare the specific antibody against the corresponding native protein.

关 键 词：表位肽 噬菌体展示 抗体 

分 类 号：Q511[生物学—生物化学]