摘要
目的 观察抗体分子通过丝状噬菌体主要外壳蛋白Ⅷ和次要外壳蛋白Ⅲ在噬菌体表面呈现效果的差异。方法 分别构建通过次要外壳蛋白Ⅲ或主要外壳蛋白Ⅷ展示抗乙肝表面抗原Fab、ScFv和角蛋白Fab的表达载体 ,制备噬菌体抗体 ,比较其抗原结合活性和Fab呈现水平 ,用多种方法尝试提高主要外壳蛋白Ⅷ介导的噬菌体展示效果。结果 主要外壳蛋白Ⅷ对不同特异性抗体Fab段和不同形式的小分子抗体 (Fab和ScFv)的展示效果均低于次要外壳蛋白Ⅲ的展示 ,增加主要外壳蛋白Ⅷ Fab对野生型和主要外壳蛋白Ⅷ的表达比例、试用不同菌株、在Fab和主要外壳蛋白Ⅷ之间插入间隔序列、换用控制更为严密的启动子等措施 ,均未能改善主要外壳蛋白Ⅷ介导的噬菌体展示。结论 以前所报道的通过主要外壳蛋白Ⅷ多价展示Fab段不是普遍存在的现象 ,对有些抗体基因不能达到多价展示。
Objective To evaluate the cpⅧ and cpⅢ mediated phage display of antibody fragments in phage antibodies. Methods Vectors displaying anti HBS Fab, ScFv or anti keratin Fab via cpⅧ orcpⅢ were constructed. Phage antibodies thereof were prepared. Antigen binding activity and Fab displaying level were tested by ELISA. Different ways were tried to improve the display efficiency of cpⅧ. Results Both antigen binding activity and Fab expression level were lower for cpⅧ constructs than cpⅢ. Efforts to increase the Fab display via cpⅧ by increasing the ratio of cpⅧ Fab fusion to wild type cpⅧ, using different E coli strains, adding spacer between Fd and cpⅧ, and using more tightly controlled promoter for cpⅧ Fab fusion were tried without succees. Conclusion Previously reported multi valence display with cpⅧ is not a general phenomenon. A more thorough understanding of the structure and function of cpⅧ is necessary before multivalence display can be obtained.\;
出处
《海军总医院学报》
2002年第2期71-74,共4页
Journal of Naval General Hospital of PLA
基金
全军十五重点课题项目 (No .0 1Z0 15 )
