抗人白细胞介素15中和抗体识别抗原表位的噬菌体肽库筛选与鉴定  

Screening and Identification of Antigen Epitope of hIL-15 Recognized by Monoclonal Antibody with 7^(TM)Phage Display Peptide Library

作　　者：高源[1,2] 郝强[1,2] 王舒宁[1,2] 张伟 薛晓畅[1,2] 

机构地区：[1]肿瘤生物学国家重点实验室 [2]第四军医大学药学系生物制药学教研室,陕西西安710032

出　　处：《生物技术通讯》2015年第2期182-185,共4页Letters in Biotechnology

基　　金：国家自然科学基金(81001182)

摘　　要：目的:用噬菌体呈现随机七肽库筛选能与抗人白细胞介素15(h IL-15)中和抗体特异性结合的模拟抗原表位肽,并初步鉴定其免疫原性。方法:以抗h IL-15中和抗体为靶分子,用生物淘洗法从噬菌体呈现线性七肽库中筛选与之结合的噬菌体克隆,用噬菌体ELISA和竞争抑制ELISA鉴定阳性噬菌体克隆;化学合成筛选得到的多肽,并与匙孔血蓝蛋白(KLH)偶联免疫BALB/c小鼠,检测其免疫原性。结果:经过3轮体外筛选后随机挑取50个阳性噬菌体克隆,ELISA检测结果显示其中15个克隆与抗h IL-15抗体有较强的结合能力,DNA测序结果得到的结构相似群为MTPFWQK、MSPFNQK、MIPYWQK和MIPFHQK;竞争性ELISA结果显示4个序列均能与IL-15竞争性地结合抗IL-15单抗;小鼠免疫实验结果显示4组多肽均能诱导IL-15特异性免疫反应。结论:筛选得到能与抗h IL-15中和抗体特异性结合,且具有免疫原性的模拟抗原7肽序列,为进一步开发h IL-15相关的多肽疫苗提供了依据。Objective： To screen out short peptide antigen epitopes capable of binding with anti human interlukin 15（hIL-15） monoclonal antibody from 7TMphage display peptide library. Methods： Using anti-hIL-15 antibody as the target protein, peptides which could bind with anti-hIL-15 antibody were selected from 7TMphage display peptide library by biopaning and identified by sandwich ELISA and competitive ELISA. The selected peptide was conjugated to keyhole limpet hemocyanin and the immunogenicity of the peptide vaccines was examined in BALB/c mice. Results： 15 of 50 phage clones were identified as positive clones which can bind to anti-hIL-15 antibody after screening and ELISA. The amino acid sequences of the positive clones are MTPFWQK, MSPFNQK, MIPYWQK, and MIPFHQK. Sera from the vaccinated mice demonstrated high-titer specific antibodies to the peptide conjugates. Conclusion： The selected peptide epitopes could bind specifically to the anti-hIL-15 antibody and induced anti-IL-15 specific immune response. The work provided the potential for developing short peptide IL-15 vaccines.

关 键 词：噬菌体随机肽库 白细胞介素15 单克隆抗体 表位 免疫 

分 类 号：R392.1[医药卫生—免疫学]