摘要
目的 通过噬菌体随机肽库筛选与白细胞介素 5 (IL 5 )高亲和力结合的相关多肽 ,观察这些多肽是否能够抑制IL 5的生物功能。方法 以重组人IL 5作为筛选分子 ,应用M13噬菌体PⅢ呈现随机 7肽库进行筛选 ,经ELISA进一步鉴定噬菌体克隆与IL 5的亲和力。阳性噬菌体克隆呈现多肽和人工合成多肽对IL 5介导的嗜酸细胞 (Eos)存活的影响。结果 经过 3轮淘筛后 ,经鉴定得到9个阳性噬菌体克隆能与IL 5呈高亲和力结合 ,其中 3个具有抑制IL 5介导的Eos存活延长作用。选择抑制作用最强的呈现多肽进行人工合成 ,发现其中一个合成多肽能够诱导Eos凋亡 ,抑制IL 5介导的存活延长作用 ,但作用强度不如噬菌体呈现多肽。结论 通过噬菌体肽库能够筛选到抑制IL 5功能的相关多肽 ,为进一步研究抗IL 5的分子药物奠定了基础。
Objective To find related polypeptides which could bind to interleukin (IL)-5 and inhibit the biologcial funtion of IL-5. Methods The recombiant human IL-5 was biotinylated with NSH-LC-Biotin, and used to react with random peptide library displaying 7 amino acids fused on protein Ⅲ of M13 phage for three rounds biopanning. The selected clons were assayed by ELISA. The displayed polypeptides of positive phage clones with high affinity and synthesized polypeptides were tested for their effects on IL-5-mediated eosinophils survival. Results Nine positive phage clones with high affinity were obtained after 3 rounds of biopanning, 3 of which could inhibit the effect of IL-5-mediated eosinophils survival. The two of most effective displayed-polypeptide were synthesized, and one of them presented the inhibition effect on IL-5 funtion in vitro by inducing apoptosis. Conclusion Potential polypeptides against IL-5 could be selected from phage display peptide library, which provide the basis for novel anti-IL-5 agents.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第5期360-364,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目 (批准号 :3 0 0 70 3 3 7)
关键词
噬菌体
随机肽库
拮抗IL-5
抑制多肽
抗体
Phage random peptides library
Interleukin-5
Polypeptide
Eosinophils
Apoptosis
