基础医学与临床 ›› 2014, Vol. 34 ›› Issue (5): 628-632.

• 研究论文 • 上一篇    下一篇

新型CD20核酸适配体的筛选及鉴定

张李钰1,胡燕1,曹蓓蓓2,段金虹1,刘喆3,杨先达1   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院北京协和医学院基础医学研究所
    3. 中国医学科学院北京协和医院
  • 收稿日期:2013-12-26 修回日期:2014-03-13 出版日期:2014-05-05 发布日期:2014-04-28
  • 通讯作者: 杨先达 E-mail:ayangmd@gmail.com
  • 基金资助:
    科技部重大科学计划

Selection and Identification of a Novel DNA aptamer against CD20 molecule

  • Received:2013-12-26 Revised:2014-03-13 Online:2014-05-05 Published:2014-04-28
  • Contact: Xian-da YANG E-mail:ayangmd@gmail.com

摘要: 目的 研发能特异性识别CD20分子的DNA适配体,使其作为肿瘤靶向分子为非霍奇金淋巴瘤的新型靶向治疗奠定基础。方法 体外合成含有21个随机寡核苷酸、全长为59个碱基的DNA文库,以CD20胞外表位肽为靶标,利用指数富集的配体系统进化技术(SELEX),从文库中筛选CD20适配体;利用流式细胞术评估适配体的富集程度、适配体与CD20多肽,适配体与CD20阳性细胞的结合特性;用MFold软件预测适配体预计结构。结果 经多轮筛选获得了能识别CD20结构的全新DNA适配体CE4-1;它能特异性地识别CD20阳性淋巴瘤细胞,与白蛋白及CD20阴性细胞的结合较弱,与经胰蛋白酶处理的CD20阳性细胞的结合也减弱。 结论 该新型适配体能选择性识别CD20结构及CD20阳性细胞,在CD20阳性淋巴瘤的靶向诊疗方面具有应用潜能。

关键词: 适配体, SELEX, CD20

Abstract: Objective To develop a CD20 aptamer that may potentially serve as a tumor-homing ligand for targeted therapy against Non-Hodgkin’s lymphoma (NHL). Methods A single-stranded 59nt DNA library containing 21nt random oligonucleotides was synthesized. A new CD20 aptamer termed CE4-1 was developed with SELEX technique, using a CD20 epitope as the target. Flow cytometry was performed to monitor the enrichment of the selected DNA pool, and the binding properties of CE4-1 towards CD20 structure and CD20-positive cells. The structure of CE4-1 was predicted by MFold software. Results The DNA aptamer CE4-1 could selectively bind with CD20 structure and CD20-positive cells, with minimal cross reactivity to BSA and CD20-negative cells. Additionally, trypsin treatment greatly reduced the binding of CE4-1 to CD20-positive cells. Conclusions A novel CD20 aptamer CE4-1 could recognize CD20 structure and CD20-positive cells selectively, which may have application potentials in targeted therapy against the CD20-positive tumors.

Key words: aptamer, SELEX, CD20

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