本文選題：噬菌體展示庫　切入點(diǎn)：腫瘤壞死因子　出處：《中國生物工程雜志》2017年05期 　論文類型：期刊論文

【摘要】：腫瘤壞死因子alpha的拮抗劑是治療多種炎癥性自身免疫疾病的首選,但抗體類拮抗物因副作用明顯而使用受限,尤其是機(jī)體內(nèi)抗抗體的產(chǎn)生,嚴(yán)重影響治療效果和藥物代謝。而肽類物除免疫原性低之外,和小分子相比也有更低的毒性和更強(qiáng)的靶標(biāo)特異性。使用7肽和12肽兩種M13噬菌體展示庫篩選TNFα拮抗肽,以分析7肽和12肽分別作為TNFα拮抗肽的親和性與功能性。經(jīng)過3~4輪的篩選和驗(yàn)證,得到2條7肽序列和2條12肽序列。利用ELISA方法檢測(cè)合成肽與TNFα結(jié)合的親和性,編號(hào)632的7肽親和性最強(qiáng),Kd=138nmol/L;編號(hào)636的12肽親和性稍差,Kd=8.59μmol/L。InsightⅡ軟件分別分析632肽和636肽與TNFα二聚體結(jié)合,發(fā)現(xiàn)632肽與TNFα二聚體結(jié)合更加穩(wěn)定,并且在細(xì)胞水平上632肽拮抗TNFα活性功能比636肽更強(qiáng),有632肽存在的條件下TNFα誘導(dǎo)的L929細(xì)胞生存率上升了3倍,而636肽的作用只有2倍。7肽比12肽更適合作為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 偽 antagonistic peptides. The affinity and function of 7 peptide and 12 peptide as antagonistic peptides of TNF 偽 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 偽 was detected by ELISA method. The binding of 632 peptide and 636 peptide to TNF 偽 dimer was found to be more stable than that of TNF 偽 dimer, and 632 peptide had the strongest affinity to TNF 偽 138nmol / L, and 636 peptide had lower affinity to TNF 偽 by using the software of TNF 偽 8.59 渭 mol/L.Insight 鈪，

本文編號(hào)：1579995