高等学校化学学报

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抗体和寡核苷酸双标记纳米金生物探针的制备及生物学特性

李富荣1, 乔飞燕2, 孔小丽1, 周汉新1, 齐晖1, 任莉莉1   

    1. 暨南大学第二临床医学院(深圳人民医院)临床医学研究中心, 深圳 518020;
    2. 兰州大学化学化工学院, 兰州 730000
  • 收稿日期:2008-04-23 修回日期:1900-01-01 出版日期:2009-01-10 发布日期:2009-01-10
  • 通讯作者: 李富荣

Preparation and Biological Characterization of Antibody and DNA Dual-labeled Gold Nanoparticles

LI Fu-Rong1*, QIAO Fei-Yan2, KONG Xiao-Li1, ZHOU Han-Xin1, QI Hui1, REN Li-Li1   

    1. Clinic Medical Research Center, The 2nd Clinic Medical College(Shenzhen People’s Hospital), Jinan University, Shenzhen 518020, China;
    2. Department of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China
  • Received:2008-04-23 Revised:1900-01-01 Online:2009-01-10 Published:2009-01-10
  • Contact: LI Fu-Rong

摘要: 纳米金通过静电吸附抗体, 与寡核苷酸共价结合制备双标记纳米金生物探针, 比较了双标记纳米金生物探针和单标记抗体IgG或ss-DNA的稳定性和反应性. 结果表明, 在水溶液中纳米金由于ss-DNA的结合使IgG抗体的吸附能力明显改善, IgG的吸附也影响二硫苏糖醇(DDT)对ss-DNA的解离作用. 双标记纳米粒上覆盖(50±15)条ss-DNA和(10±2)条IgG, 较单标记ss-DNA纳米金上的(70±15)条要少. 斑点免疫和杂交实验证明, 纳米金表面标记的IgG和ss-DNA具有良好生物学活性. 双标记纳米金生物探针在超微量蛋白质的检测中具有应用价值.

关键词: 金纳米粒子, 生物探针, 寡核苷酸, 抗体

Abstract: Gold nanoparticles labeled by both antibody(IgG) and single stranded DNA(ss-DNA) were synthesized and characterized. The stability and reactivity of the dual-labeled nanoparticles were compared with the conventional IgG or ss-DNA modified nanoparticles. It was found that the IgG adsorption significantly improved the stability of the nanoparticles in aqueous solution, which is beneficial for attaching ss-DNA. The presence of IgG also effectively prohibits the desorption of ss-DNA against dithiothreitol(DTT) displacement. The coverage on dual-labeled nanoparticles was found to be (50±15) ss-DNA/nanoparticle and (10±2) IgG/nanoparticle, respectively, compared to the value of (70±15) ss-DNA/nanoparticle of only ss-DNA-labeled gold nanoparticles. Dot-immuno and cross-linking experiments confirmed that both the IgG and ss-DNA retained their bioactivity on the nanoparticle surface. The dual-labeled nanoparticles have potential to be used as novel bio-probes for ultrasensitive detection.

Key words: Gold nanoparticle, Bio-probes, DNA, Antibody

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