摘要
目的 :利用噬菌体肽库获得脂多糖的模拟表位。方法 :用脂多糖免疫大耳白兔获得抗血清 ,从中纯化抗脂多糖的多克隆抗体。以纯化的抗体IgG为筛选靶分子 ,对噬菌体随机十二肽库进行亲和筛选。经 3轮筛选后 ,随机挑取 2 5个克隆测序 ,并进行噬菌体ELISA结合实验及脂多糖竞争抑制实验 ,以确定阳性克隆。结果 :获得 12种序列 ,ELISA实验结果显示其中 6个为阳性克隆 ,且经脂多糖竞争抑制实验进一步证实。结论 :获得了 6个脂多糖的模拟表位 ,为脂多糖抗原表位的研究及疫苗研究提供了线索。
Objective:To obtain mimotopes of LPS by phage_displayed random peptide library.Methods:Polyclonal IgG antibody was obtained by immunizing rabbits and anti_sera were purified. The specific polyclonal antibody was used as the target for biopanning of a 12_mer phage_displayed random peptide library. After 3 rounds of screening, 25 clones were randomly selected for DNA sequencing, then detected by ELISA and LPS competition experiments to identify the positive clones.Results:Twelve sequences were obtained. Six of twelve clones were proved to be positive, and confirmed by competition experiments.Conclusion:The six phage_displayed peptides as mimotope of LPS were obtained. The findings provide a clue to LPS epitope mapping and vaccine development. [
出处
《军事医学科学院院刊》
CSCD
北大核心
2003年第4期271-273,共3页
Bulletin of the Academy of Military Medical Sciences
关键词
脂多糖
噬菌体随机肽库
模拟表位
筛选
lipopolysaccharide
phage_displayed random peptide library
mimotope
