噬菌体表达短肽模拟脂多糖类脂A表位的研究

Preparation of peptides mimicring lipid A epitope using phage display peptide library

目的：得到拟类脂Ａ的六肽。方法：用抗类脂Ａ的单克隆抗体筛选噬菌体随机六肽库，经四轮筛选后进行ＥＬＩＳＡ检测，用得到的１５个阳性克隆分别免疫小鼠，并将使小鼠产生抗ＬＰＳ抗体的１０个克隆进行测序，将保守序列进行肽合成。结果：得到保守序列为ＬｙｓＰｈｅＳｅｒＳｅｒＡｒｇ，即ＫＦＳＳＲＸ。固相合成此小肽可与其１Ｂ６抗体结合，且此结合可被ＬＰＳ抑制。

Objective:Finding mimic Lipid A 6 mer peptides.Methods:Used anti Lipid A McAb to fish out phage display 6 mer peptide random library.After 4 rounds selection,15 positive clones were detected by ELISA and used to immunize mice.Ten of them could get antibody for LPS.To analyze the DNA sequences and to synthesize the peptide from conservative sequences.Results:The conservative amino acid sequence is Lys Phe Ser Ser Arg or KFSSRX.The solid phase synthesis peptide can bind to 1B6 antibody and this binding can be inhibited by LPS.Conclusion:These results suggest that 6 mer peptide can mimicry the epitope of LPS or Lipid A.