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Bioelectrochemical-Array Sensor Using DNA Conjugate as Chemical Sensing Element

使用 DNA 缀合物作为化学传感元件的生物电化学阵列传感器

基本信息

批准号：
15550073
负责人：
NAKANO Koji
金额：
$ 2.37万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15550073/
关键词：
DNA conjugate gene sensor DNA microarray SECM high-throughput assay

项目摘要

This research project has involved 1)enzymic synthesis of poly(hydroquinone), PQ, 2)chemically modified electrode preparations and gene sensor applications, and finally, 3)scanning electrochemical microscopic(SECM) imaging of DNA microarray. First, we synthesized the redox polymer by enzymic oxidation of albutin to form poly(hydroquinone glycoside) derivative, which was subsequently converted to PQ by chemical removal of the sugar-protecting group. The polymer was then applied to DNA-conjugate modified electrode having a single-stranded oligodeoxyribonucleotide (ODN)/PQ surface structure. Electrochemical studies showed that the electrode was responsive to the hybridization of the surface-anchored ODN by changing its redox activity. By detailed studied on the electrochemistry and the bioaffinity reaction, the DNA-conjugate modified electrode was concluded to be a candidate of electrochemical sensor capable of detection of specific genes. As the start point of high-throughput analytical device development, we made an attempt of two-dimensional imaging of the specific electrochemical activity of the DNA conjugate electrode using a SECM. We have finally succeeded 1)2D imaging of the DNA microdots (20 micron) and 2)detection of DNA hybridization through 2D imaging by monitoring the redox activity of the microdot.

本课题主要研究内容包括：1)聚对苯二酚的酶促合成，PQ, 2)化学修饰电极的制备和基因传感器的应用，3)DNA微阵列扫描电化学显微镜（SECM）成像。首先，我们通过酶氧化白蛋白合成氧化还原聚合物，形成聚对苯二酚糖苷衍生物，随后通过化学去除糖保护基团转化为PQ。然后将该聚合物应用于具有单链寡脱氧核糖核苷酸(ODN)/PQ表面结构的dna共轭修饰电极。电化学研究表明，电极通过改变ODN的氧化还原活性来响应表面锚定的ODN的杂交。通过详细的电化学和生物亲和反应研究，认为dna共轭修饰电极是一种能够检测特定基因的电化学传感器候选材料。作为高通量分析装置开发的起点，我们尝试使用SECM对DNA共轭电极的比电化学活性进行二维成像。我们最终成功实现了1)DNA微点（20微米）的二维成像，2)通过监测微点的氧化还原活性，通过二维成像检测DNA杂交。