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目的 利用噬菌体随机 15肽库确定SLE相关多肽。方法 从系统性红斑狼疮 (SLE)病人血清中纯化抗dsDNA多克隆抗体 ,以此为筛选配基 ,对噬菌体随机 15肽库进行亲和筛选。结果经 3轮筛选 ,每轮的投入 /产出比逐轮升高 ,提示筛选具有良好的富集效果。第 3轮筛选后选其中 18个克隆进行结合试验 ,结果显示有 6个仅与SLE病人血清反应而不与正常人血清反应 ,经DNA序列测定 ,发现其中 5个克隆序列相同 ,通过该序列推出插入的外源短肽序列 ,合成并纯化该肽段。用此肽段分别与 2 0例SLE病人及 2 0例正常人血清反应 ,结果差异有显著性。结论 该肽段可能是与SLE相关的抗原多肽。
Objective To identify SLE-related polypeptides using a phage random 15-peptide library. Methods The anti-dsDNA polyclonal antibody was purified from the serum of patients with systemic lupus erythematosus (SLE), which screened the ligand for the random selection of phage 15-peptide library. Results After three rounds of screening, the input / output of each round was higher than that of rounds, suggesting that screening had a good enrichment effect. Eighteen clones were screened after the third round of screening. The results showed that six of them reacted only with sera of SLE patients but not normal sera, and the DNA sequence analysis showed that the five clones had the same sequence, and the sequence was introduced The inserted exogenous short peptide sequence is synthesized and purified. With this peptide were 20 cases of SLE patients and 20 normal serum reaction, the results were significant differences. Conclusion The peptide may be related to SLE antigen polypeptide.