摘要
利用噬菌体抗体库筛选技术获得抗雄性特异性抗原的噬菌体Fab抗体,首次采用雄鼠脾细胞对鼠源性抗雄性特异性抗原噬菌体Fab抗体库进行3轮亲和富集和2轮雌鼠脾细胞吸附,对筛选后特异性噬菌体Fab抗体进行ELISA分析,重组率鉴定及基因测序分析。结果显示,5次筛选后的15个菌落中有9个能产生抗雄性特异性抗原特异性噬菌体抗体,噬菌体Fab抗体的基因重组率为60%,E5克隆的重链、轻链可变区序列分别属于VH1和VκⅣ基因家族,这为挑选出高亲和力的抗雄性特异性抗原噬菌体Fab抗体奠定了实验基础,将推进雄性特异性抗原及其抗体的研究进程,并为性别控制研究开创新途径。
To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60% . Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vκ had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.
出处
《生物工程学报》
CAS
CSCD
北大核心
2006年第5期727-732,共6页
Chinese Journal of Biotechnology
关键词
小鼠
H-Y抗原
噬菌体展示技术
抗体库
mouse, H-Y antigen, phage display technology, antibody library
