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目的 利用噬菌体六肽库筛选肿瘤坏死因子 (TNF)受体的短肽配体。方法 利用可溶性TNF受体Ⅰ (sTNFRⅠ )蛋白筛选噬菌体随机六肽库 ,经过 4轮筛选和酶联免疫吸附测定 ,获得能和sTNFR相结合的噬菌体克隆。对部分阳性噬菌体克隆进行DNA测序 ,推算出相应的氨基酸序列 ,并进行保守性分析。按其中的保守序列进行多肽设计、合成 ,通过细胞学实验鉴定其生物活性。结果 得到 30个可模拟TNF的阳性克隆 ,对其中 6个进行单链DNA测序和分析。合成和纯化短肽 (八肽 )WH70 1,体外细胞毒实验结果证实了该序列的有效性。结论 获得了可模拟TNF受体亲和力的短肽配体 ,为研制新的TNF受体显像多肽配基提供了实验依据。
Objective To screen short peptide ligand of tumor necrosis factor (TNF) receptor by phage hexapeptide library. Methods The phage random hexapeptide library was screened by soluble TNF receptor Ⅰ (sTNFR Ⅰ) protein. After 4 rounds of screening and enzyme - linked immunosorbent assay (ELISA), phage clones capable of binding to sTNFR were obtained. DNA sequencing of some positive phage clones, deduced the corresponding amino acid sequence, and conduct conservative analysis. Polypeptides were designed and synthesized according to the conserved sequences, and their biological activities were identified by cytological experiments. Results 30 positive clones that could simulate TNF were obtained, of which 6 were single-stranded DNA sequencing and analysis. Synthesis and purification of short peptides (octapeptide) WH70 1, in vitro cytotoxicity test results confirmed the validity of the sequence. CONCLUSIONS Short peptide ligands that mimic the affinity of TNF receptors were obtained and provided an experimental basis for the development of novel ligands for TNF receptor imaging peptides.