中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (33): 3107-3115.doi: 10.3969/j.issn.1673-5374.2013.33.004

• 原著:退行性病与再生 • 上一篇    下一篇

人源噬菌体抗体库的构建及抗beta-淀粉样蛋白寡聚体单链抗体的筛选

  

  • 收稿日期:2013-04-25 修回日期:2013-07-01 出版日期:2013-11-25 发布日期:2013-11-25

Construction of human Fab library and screening of a single-domain antibody of amyloid-beta 42 oligomers

Zuanning Yuan1, 2, Minge Du1, 2, Yiwen Chen3, Fei Dou1, 2   

  1. 1 Key Laboratory of Cell Proliferation and Regulation Biology, Ministry of Education, Beijing Normal University, Beijing 100875, China
    2 Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Southeast University, Nanjing 210009, Jiangsu Province, China
    3 Geriatric Internal Department of Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China
  • Received:2013-04-25 Revised:2013-07-01 Online:2013-11-25 Published:2013-11-25
  • Contact: Fei Dou, Ph.D., Professor, Key Laboratory of Cell Proliferation and Regulation Biology, Ministry of Education, Beijing Normal University, Beijing100875, China; Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Southeast University, Nanjing 210009, China, douf@bnu.edu.cn.
  • About author:Zuanning Yuan, Master.

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摘要：

通过人源噬菌体抗体库筛选人源抗体被认为是一种高效快速的beta-淀粉样蛋白寡聚体途径。鉴于此，实验首先体外合成beta-淀粉样蛋白(1-42)寡聚体，并构建了基于6个正常人血液样品的噬菌体抗体库。最后对此噬菌体抗体库进行3轮体外结合-洗脱-扩增筛选后发现了抗beta-淀粉样蛋白寡聚体的人单链抗体。Western Blot及ELISA检测结果显示，此抗体可与有毒性的beta-淀粉样蛋白四聚体或九寡聚体特异性结合，而不与beta-淀粉样蛋白单体及大分子聚体结合。实验成功构建了人源噬菌体抗体库并筛选了抗Aβ寡聚体单链抗体，可以特异识别Aβ寡聚体。