7肽和12肽两种M13噬菌体展示库筛选肿瘤坏死因子alpha拮抗肽的比较

发布时间：2018-03-07 16:09

  本文选题：噬菌体展示库　切入点：肿瘤坏死因子　出处：《中国生物工程杂志》2017年05期 　论文类型：期刊论文

【摘要】：肿瘤坏死因子alpha的拮抗剂是治疗多种炎症性自身免疫疾病的首选,但抗体类拮抗物因副作用明显而使用受限,尤其是机体内抗抗体的产生,严重影响治疗效果和药物代谢。而肽类物除免疫原性低之外,和小分子相比也有更低的毒性和更强的靶标特异性。使用7肽和12肽两种M13噬菌体展示库筛选TNFα拮抗肽,以分析7肽和12肽分别作为TNFα拮抗肽的亲和性与功能性。经过3~4轮的筛选和验证,得到2条7肽序列和2条12肽序列。利用ELISA方法检测合成肽与TNFα结合的亲和性,编号632的7肽亲和性最强,Kd=138nmol/L;编号636的12肽亲和性稍差,Kd=8.59μmol/L。InsightⅡ软件分别分析632肽和636肽与TNFα二聚体结合,发现632肽与TNFα二聚体结合更加稳定,并且在细胞水平上632肽拮抗TNFα活性功能比636肽更强,有632肽存在的条件下TNFα诱导的L929细胞生存率上升了3倍,而636肽的作用只有2倍。7肽比12肽更适合作为TNFα拮抗肽。
[Abstract]:The antagonist of tumor necrosis factor (alpha) is the first choice in the treatment of various inflammatory autoimmune diseases, but the use of antibody antagonists is restricted due to obvious side effects, especially the production of anti-antibodies in the body. In addition to its low immunogenicity, peptides have lower toxicity and stronger target specificity than small molecules. Two M13 phage display libraries of 7 and 12 peptides were used to screen TNF 伪 antagonistic peptides. The affinity and function of 7 peptide and 12 peptide as antagonistic peptides of TNF 伪 were analyzed. After 3 rounds of screening and verification, two 7 peptide sequences and 2 12 peptide sequences were obtained. The affinity of synthetic peptides to TNF 伪 was detected by ELISA method. The binding of 632 peptide and 636 peptide to TNF 伪 dimer was found to be more stable than that of TNF 伪 dimer, and 632 peptide had the strongest affinity to TNF 伪 138nmol / L, and 636 peptide had lower affinity to TNF 伪 by using the software of TNF 伪 8.59 渭 mol/L.Insight 鈪，

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