文化大學機構典藏 CCUR:Item 987654321/24305
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    jsp.display-item.identifier=請使用永久網址來引用或連結此文件: https://irlib.pccu.edu.tw/handle/987654321/24305


    题名: DEVELOPMENT OF A SURFACE PLASMON RESONANCE-BASED DNA SENSOR AND ITS APPLICATION FOR SCREENING DNA-TARGETED ANTICANCER DRUGS
    作者: Tsai, WC (Tsai, Wen-Chi)
    Hsu, YH (Hsu, Yung-Hui)
    贡献者: Grad Inst Biotechnol
    关键词: DNA hybridization
    DNA/small-molecule interaction
    Mixed self-assembled monolayer
    Surface plasmon resonance
    日期: 2012
    上传时间: 2013-02-25 14:36:29 (UTC+8)
    摘要: In this paper, we present a surface plasmon resonance (SPR)-based sensor for measuring DNA hybridization and DNA/small-molecule interactions. A mixed self-assembled monolayer (SAM) was used to optimize the biosensor sensitivity. It was observed that the mixed SAM formed by mixing 10 mM of 16-mercapto-1-hexadecanoic acid (16-MHA) and 6-mercapto-1-hexanol (6-MCH) at a 1: 10 molar ratio showed the best results. Subsequently, avidin was attached to the carboxyl groups on the SAM to serve as a binding element for biotinylated single-stranded (ss) DNA. The ssDNA-coated sensor was first evaluated as a nucleic acid biosensor through a DNA-DNA hybridization assay for synthetic 28-mer ssDNA. A linear calibration curve was observed in the range of 0.25-2.5 mu g/mL. Non-complementary DNA induced no significant SPR angle shift, which demonstrated the specificity of the assay. Secondly, the sensor was used to monitor the binding kinetics of DNA/small-molecule interactions in real time. The dissociation constant between immobilized DNA and sanguinarine was determined to be 8.0 x 10(-6) M. This complies with most data from the literature. In addition, the sensor could be regenerated with 0.01 M HCl and would be feasible for multiple testing. In conclusion, the experimental approach described in this study allows analysis of molecular interactions between DNA-binding drugs and selected targeted DNA sequences.
    關聯: ANALYTICAL LETTERS 卷: 45 期: 11 頁數: 1495-1505
    显示于类别:[生物科技研究所 ] 期刊論文

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