Objective By using phage-displayed 12 peptide libraries to select antagonists to TNF-α which could inhibit TNF-α-induced cytotoxicity in vitro.Methods A random 12-mer peptide library was screened with 1 ml rhTNF-α (100 μg),according to biopanning procedure of New England Biolabs Inc.Combinant human TNF-α (2.2×108 U/ml) were purchased from Biotechnology Centre.The mouse fibroblast cell L929 was used to measure TNF-α-mediated cytotoxicity.DNA was sequenced on an ABI PRISM automated 377 DNA sequencer.Results Randomly selected phage clones after the third biopanning were tested for their special binding with TNF-α in ELISA.The 21 positive clones were sequenced with an ABI PRISM Dye terminator cycle sequencing reaction kit.From the 21 clones tested,one (11),consistently inhibited the TNF-α-induced cytotoxicity on L929 cells.The inhibition was dose-dependent at concentration of TNF-α giving 38.2%.No inhibition was observed with a control phage.Conclusion These sequences may take for the basis of sythesis of small peptide antagonist to TNF-α.%目的从噬菌体随机多肽库中筛选出与肿瘤坏死因子-α(TNF-α)特异结合，并能阻断其生物活性的小肽，为建立一种新型的抗TNF-α多肽药物奠定基础。方法用突变体TNF-α和天然型TNF-α分别作为靶蛋白，用噬菌体随机多肽库进行多次淘筛，经序列测定及体外酶联免疫吸附法(ELISA)实验选出候选多肽，进行抗TNF-α诱导的L929细胞毒作用。结果筛选到的二个序列不同但有一定相关性的候选小肽，其中由天然型TNF-α作为靶蛋白筛选出的12肽具有抗TNF-α诱导的L929细胞毒作用。结论这些序列可以做为合成小肽TNF-α活性拮抗剂的依据。
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