CAJ | 학술논문

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（hIL-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hIL-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hIL-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hIL-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hIL-15相关的多肽疫苗提供了依据。
목적：용서균체정현수궤칠태고사선능여항인백세포개소15（hIL-15）중화항체특이성결합적모의항원표위태，병초보감정기면역원성。방법：이항hIL-15중화항체위파분자，용생물도세법종서균체정현선성칠태고중사선여지결합적서균체극륭，용서균체ELISA화경쟁억제ELISA감정양성서균체극륭；화학합성사선득도적다태，병여시공혈람단백（KLH）우련면역BALB/c소서，검측기면역원성。결과：경과3륜체외사선후수궤도취50개양성서균체극륭，ELISA검측결과현시기중15개극륭여항hIL-15항체유교강적결합능력，DNA측서결과득도적결구상사군위MTPFWQK、MSPFNQK、MIPYWQK화MIPFHQK；경쟁성ELISA결과현시4개서렬균능여IL-15경쟁성지결합항IL-15단항；소서면역실험결과현시4조다태균능유도IL-15특이성면역반응。결론：사선득도능여항hIL-15중화항체특이성결합，차구유면역원성적모의항원7태서렬，위진일보개발hIL-15상관적다태역묘제공료의거。
Objective: To screen out short peptide antigen epitopes capable of binding with anti human inter?lukin 15(hIL-15) monoclonal antibody from 7TMphage display peptide library. Methods: Using anti-hIL-15 anti?body as the target protein, peptides which could bind with anti-hIL-15 antibody were selected from 7TMphage dis?play 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 pep?tide 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.