利用噬菌体随机肽库筛选IL-5结合的相关多肽  

Potential polypeptides recognized by interleukin(IL)-5 through screening phage displayed random peptide library

作　　者：赖克方[1] 孙宝清[1] 黄海鹭[1] 钟南山[1] 

机构地区：[1]广州呼吸疾病研究所,广州510120

出　　处：《中国免疫学杂志》2004年第7期483-486,共4页Chinese Journal of 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)

摘　　要：目的 :采用噬菌体随机肽库筛选人IL 5 ,以获得能高亲和力结合IL 5的相关多肽。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。经过三轮淘筛后 ,经夹心ELISA、竞争ELISA法进一步鉴定噬菌体克隆与IL 5的亲和力 ,对阳性克隆进行了扩增和DNA测序 ,据此推导结合随机多肽的氨基酸序列。结果 :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。通过测序和序列比较分析 ,发现CX1 2 AS为相对保守的结合表位。结论 :研究表明通过噬菌体肽库能够筛选到IL 5结合的相关多肽 ,为进一步研究ILObjective:To find related polypeptides which could bind interleukin (IL) 5 with high affinity.Methods:The recombinant human IL 5 was biotinylated with NSH LC Biotin,then biotinylated IL 5 was used to react with random peptide library displaying 7 amino acids fused on protein III of M13 Phage for three rounds biopanning.The selected clones were assayed by sandwich enzyme linked immunosorbent assay(ELISA)and competition ELISA.The positive clones with high affinity were used for automated sequencing with dye labeled dideoxynucleotides,and the amino acid sequence of polypeptide displayed on phage was deduced.Results:The enrichment was shown by ELISA after 3 rounds of biopanning.9 positive clones could bind to IL 5 with high affinity.Sequencing of the genes encoding these peptides on 9 positive clones showed some conserved epitope information such as SX 1 2 AS,ALAS.Conclusion:Potential polypeptides binding IL 5 with high affinity could be selected from phage display peptide library,SX 1 2 AS may be the motifs recognized by IL 5.

关 键 词：IL-5 噬菌体呈现肽库 多肽 

分 类 号：R392-33[医药卫生—免疫学]