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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。
In order to establish a new method of preparing anti-protein antibodies based on epitopes. The T7 phage was used as a display vector to display the epitope peptide of the protein on its surface. The recombinant phage was extracted as the immunogen and the titer and specificity of the antibody were detected by ELISA, Western blot and Cellular Immunofluorescence. The results showed that the antibodies against T7 phage displaying protein epitopes could specifically recognize the corresponding epitope peptide, and the antibody against Pif1-α epitope could specifically recognize Pif1-α protein. The conclusion is that T7 phage displaying small peptides on the surface is very immunogenic and, if the small peptides displayed are indeed protein epitopes, animals can be immunized as surrogate antigens for intact protein antigens to prepare antibodies that recognize intact proteins.