噬菌体随机肽库筛选日本血吸虫22.6kDa蛋白有效抗原表位的初步研究

Studies on immunoprotection against Schistosoma japonicum induced by phage display peptides mimicking the epitopes of 22.6kDa antigen of Schistosoma japonicum

目的 :从噬菌体随机 12肽库中筛选出中国大陆株日本血吸虫 2 2 .6kDa蛋白 (Sj2 2 .6kDa)抗原表位的短肽分子 ,探讨其抗血吸虫的交叉免疫保护效果。方法 :以纯化的抗日本血吸虫 2 2 .6kDa的多克隆抗体IgG为配基 ,对噬菌体 12肽库进行 5轮亲和筛选 ,富集特异性噬菌体 ,挑取克隆 ,经序列分析、免疫识别和动物初筛后获得阳性克隆。结果 :经过 5轮筛选 ,特异性噬菌体得到有效富集。随机挑选的 2 4个噬菌体克隆中获得 4个有效抗原表位。结论 :利用噬菌体随机肽库筛选可获得类似日本血吸虫 2 2 .6kDa有效抗原表位的短肽分子 ,这些短肽分子能诱导抗血吸虫的保护性免疫。

Objective To obtain the short peptides mimicking antigenic epitopes of 22.6?kDa antigen of Schistosoma japonicum(Sj) from phage display library,and explore their cross protective immunity against Sj in mice.Methods Purified polyclone IgG antibody against the Sj 22.6?kDa were used to immunoscreen a 12 mer phage display library for the epitopes of Sj 22.6?kDa.Five rounds of biopanning were carried out and 24 clones from the fifth round biopanning were random selected,identified by Western blot and sequenced.The mice were immunized with the obtained 4 positive clones.Results Four different epitopes of Sj 22.6?kDa were obstained.Conclusion Antigenic epitopes of Sj 22.6?kDa can be prepared by immunoscreening phage display library,a significant protective immunity against Sj can be induced by these epitopes in mice.