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利用噬菌体库筛选IL-6拮抗剂及功能研究
Searching interleukin 6 antagonists by screening random peptide libraries
【摘要】 目的 以人sIL 6R为靶分子 ,利用噬菌体随机 15肽库筛选其亲和短肽 ,结合计算机模拟分析 ,获得具有抑制IL 6 IL 6R结合作用的短肽序列 ,作为IL 6拮抗剂。方法 通过亲和筛选 ,获得一组sIL 6R特异亲和序列。测定序列 ,结合计算机模拟 ,选取具有潜在拮抗功能的 2条肽 (命名为pT1、pT2 )。合成短肽 ,进行ELISA竞争抑制实验和细胞增殖抑制实验。结果 pT1、pT2均可抑制IL 6 sIL 6R的结合。pT1、pT2可抑制XG 7细胞的增殖 ,并且这种抑制作用随着合成肽浓度的增高而增强 ,pT1、pT2不影响 7TD1细胞的增殖。结论 利用亲和筛选并结合计算机模拟从随机 15肽库中获得可抑制IL 6 sIL 6R结合的短肽 ,并可抑制IL 6依赖细胞XG 7的增殖。
【Abstract】 Objective Searching interleukin 6 (IL-6) antago nists by screening random peptide libraries. Methods Huma n sIL-6R was immobilized in high affinity 35mm dish and screened by 15-mer ran dom peptides displayed on the phage. The monoclonal phagemids displaying peptide s specifically against shIL-6R (soluble human IL-6 receptor) by screening were sequenced. Utilizing the homology program of the insight Ⅱ (95.5) software, we found two sequences which might be antagonists against IL-6 and named pT1 and pT2. The antagonistic reaction of the two synthetic peptides against IL-6 were determined with ELISA and the test of proliferation of XG-7 cells. R esults The results of ELISA showed that the synthetic peptides block ed the binding of IL-6 to IL-6R. The pT1 and pT2 also inhibited the proliferat ion of XG-7 cells, which came from human myeloma and proliferate depending on I L-6 and the inhibition was positively related to dose of the synthetic peptides . But the two peptides had no effect on the proliferation of murine cells 7TD1 w hich were IL-6 dependent. Conclusion The two peptides, p T1 and pT2, obtained by screening random peptide libraries, have the antagonisti c reaction against IL-6.
- 【文献出处】 中华微生物学和免疫学杂志 ,Chinese Journal of Microbiology and Immunology , 编辑部邮箱 ,2003年08期
- 【分类号】R392
- 【被引频次】3
- 【下载频次】81