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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 the mimotope of lipopolysaccharide by phage peptide library. Methods: The antiserum was obtained by immunization of large white rabbits with lipopolysaccharide and the polyclonal antibody against lipopolysaccharide was purified from it. Purified antibody IgG was screened for the target molecule, and the phage randomized 12-mer peptide library was screened by affinity. After 3 rounds of screening, randomly selected 25 clones sequenced, and phage ELISA binding experiments and lipopolysaccharide competition inhibition experiments to determine the positive clones. Results: Twelve kinds of sequences were obtained. Six of them were positive clones by ELISA, which was further confirmed by lipopolysaccharide competitive inhibition test. CONCLUSION: Six mimotopes of lipopolysaccharide were obtained, which provided a clue for the research of lipopolysaccharide epitopes and vaccine research.