利用噬菌体环肽库筛选可溶性肿瘤坏死因子受体1的特异性结合肽

Specific sTNFR 1 Binding Peptide Selected from Disulfide Constrained Phage Display Peptide Library

目的 应用噬菌体环肽库筛选肿瘤坏死因子α(TNF- α) 的模拟肽, 为研制模拟TNF -α的新型多肽药物奠定基础。方法 用亲和纯化的可溶性TNF受体1 (sTNFR -1) 作为筛选配基, 对噬菌体随机环肽库进行亲和筛选, 利用胞毒效应进行生物学筛选, 并进一步对某些阳性克隆进行序列测定和分析。结果 经3 轮筛选, 每轮的投入/产出比逐轮提高, 说明筛选具有良好的富集效果。并得到模拟跨膜型TNF -α(TM -TNF- α)、可溶型TNF -α(S- TNF -α) 的模拟肽和拮抗TNF作用的sTNFR 1 封闭肽。获得了TM -TNF -α的模拟肽的氨基酸序列的基序。结论 获得的模拟TM -TNF- α生物学效应的sTNFR -1结合肽, 可望发展成为一种新的TNF- α肽类新药。

Objective To develop a new type of tumor necrosis factor-α (TNF-α) peptide as drug,TNF-α mimic peptides were selected from disulfide constrained phage peptide library.Methods By using affinity purified soluble TNF-α receptor 1 (sTNFR1) as screening ligand,affinity selection was done from disulfide constrained phage random peptide library. Cytotoxicity was estimated for bioactivity selection. DNA from positive clones was sequenced and analyzed.Results After three rounds of affinity selection,the ratio of output verse input was increased,which indicated the efficient enrichment. Three different kinds of sTNFR1 binding peptides were obtained: TM-TNF mimic peptide,S-TNF mimic peptide and antagonized TNF peptide respectively. The motif for TM-TNF mimic peptide was obtained.Conclusion The obtained TM-TNF-α mimic peptides can be developed into one new type of TNF-α peptide drug.