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TNF α是一种在机体抗感染、抗肿瘤过程中发挥重要作用的细胞因子 ,其对机体具有保护和损伤两方面的作用 ,为了探讨新的抑制TNF α所致炎症损伤等反应的手段 ,构建了细菌鞭毛递呈的随机肽库 ,利用构建得到的肽库 ,进行TNF α特异性结合肽的筛选工作。经过 5轮筛选及DNA测序 ,共得到 6条小肽编码序列。其中 2条序列中含有一V N WG的相同序列框架。进行 6条序列与TNF α结合力的确证后 ,选择了其中的 4条肽序列进行人工化学合成、纯化及鉴定。利用L92 9细胞及MTT法对 4条小肽进行活性测定 ,检测其对TNF α的抑制活性。结果表明 ,在TNF α对L92 9细胞毒性为 30 %左右时 ,含同源序列框架的 2条肽可抑制 90 %左右的TNF α活性
TNFα is a cytokine that plays an important role in the anti-infective and antitumor activities of the body. It has both the protective and the damaging effects on the body. In order to explore a new means to inhibit the inflammatory damage induced by TNFα, The random peptide library presented by bacterial flagella was used to screen the TNFα-specific binding peptides by using the constructed peptide library. After five rounds of screening and DNA sequencing, a total of six small peptide coding sequences were obtained. Two of these sequences contain the same sequence framework of a V N WG. After confirming the binding affinity of 6 sequences to TNFα, 4 peptide sequences were selected for artificial chemical synthesis, purification and identification. The activities of four small peptides were measured by L92 9 cells and MTT assay, and their inhibitory activity on TNFα was tested. The results showed that two peptides containing the homologous sequence could inhibit about 90% of TNFα activity when the cytotoxicity of TNFα to L92 9 was about 30%