高等学校化学学报

高等学校化学学报 ›› 2011, Vol. 32 ›› Issue (11): 2509.

• 研究论文 • 上一篇    下一篇

基于核酸适体与互补核酸和目标蛋白之间竞争反应的PDGF蛋白特异性识别

郑静1,2, 秦佳华1, 龚晨1, 黄焕1, 何品刚2, 方禹之2   

  1. 1. 上海工程技术大学化学化工学院, 上海 201620;
    2. 华东师范大学化学系, 上海 200062
  • 收稿日期:2010-12-06 修回日期:2011-01-07 出版日期:2011-11-10 发布日期:2011-10-14
  • 通讯作者: 郑静 E-mail:kkzhengjing707@163.com
  • 基金资助:

    国家自然科学基金(批准号: 20675031)和上海市教委科研创新项目(批准号: 09YZ374)资助.

Study on specific recognition for PDGF based on the competitive reaction between Aptamer-DNA and Aptamer-protein

ZHENG Jing1,2*, QIN Jia-Hua1, GONG Chen1, HUANG Huan1, HE Pin-Gang2, FANG Yu-Zhi2   

  1. 1. College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China;
    2. Department of Chemistry, East China Normal University, Shanghai 200062, China
  • Received:2010-12-06 Revised:2011-01-07 Online:2011-11-10 Published:2011-10-14
  • Contact: ZHENG Jing E-mail:kkzhengjing707@163.com
  • Supported by:

    国家自然科学基金(批准号: 20675031)和上海市教委科研创新项目(批准号: 09YZ374)资助.

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被引次数

6

摘要： 报道了一种利用核酸适体与互补核酸和目标蛋白之间的竞争反应用于PDGF蛋白的特异性识别方法. 采用反相微乳液技术制备了包覆有亚甲基蓝(MB)的SiO2纳米复合物(SiO2-MB), 利用固定在磁性纳米颗粒上的核酸适体与SiO2-MB纳米颗粒标记的互补核酸进行杂交反应, 将一定数量的SiO2-MB纳米颗粒固定于磁性纳米颗粒的表面, 引入PDGF目标蛋白后, 利用核酸适体与互补核酸和PDGF之间的竞争反应, 通过检测MB电化学信号的变化来检测PDGF. 该方法对PDGF蛋白具有很高的特异性识别能力, 不受其它蛋白质如免疫球蛋白G、凝血酶和溶菌酶等的干扰. 实验结果表明, PDGF浓度在5.51×10-17~1.01×10-15 mol/L范围内具有良好的线性关系, 检出限可达1.31×10-17 mol/L.

关键词: 核酸适体, PDGF蛋白, 特异性识别

Abstract: A specific detection approach for platelet-derived growth factor(PDGF) via the competitive reaction between Aptamer-DNA and Aptamer-protein was established. In the paper, SiO2-MB nanocomposite was synthesized by reverse microemulsion method. For this proposed aptasensor, aptamer was immobilized on the magnetic nanoparticles, and the complementary oligonucleotide was labeled with SiO2-MB nanocomposite, Through the hybridization between the aptamer and its complementary oligonucleotide, a certain amount of SiO2-MB nanocomposite was gathered onto the magnetic nanoparticles. In the presence of PDGF, the aptamer was in a conformational equilibrium between Aptamer-DNA and Aptamer-protein, the aptamer prefers to bind with PDGF, resulting in a significant decrease of the electrochemical signal of MB. Upon this, we were able to detect the binding events between the aptamer and the protein. The assay was highly sensitive for PDGF, and is interference from other proteins such as IgG, thrombin and lysozyme. The linear range of the PDGF is 5.51×10-17 ～1.01×10-15mol/L with a detection limit of 1.31×10-17mol/L.

Key words: aptamer, PDGF, specific recognition

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