Molecular Design of DNA Conjugate and its Applications to Biosensing

我们根据我们独特的原理开发了新的生物传感技术，并提出了简便、有效和高灵敏度的分析方法。该原理包括通过使用携带dna的纳米颗粒的非交联聚集进行单碱基识别，这些纳米颗粒是通过接枝寡脱氧核糖核苷酸的热响应聚合物的自组装形成的。在这项研究中，我们设计了新的dna偶联材料，包括dna响应水凝胶、dna合成聚合物块型偶联物和dna连接金纳米颗粒，作为基于这一原理的基因诊断分析工具。成果总结如下：1。dna固定聚苯乙烯乳胶和dna连接金纳米颗粒的独特聚集证明了这一原理是普遍成立的。携带dna的纳米颗粒的电泳迁移率发生了序列依赖性的剧烈变化。采用合理设计的单链DNA作为交联剂的新型杂化水凝胶，可根据样品DNA序列收缩或膨胀，从而精确区分单碱基差异。采用DNA合成聚合物嵌段型偶联物作为亲和配体，建立了分离单链样品DNA及其点突变体的亲和毛细管电泳技术。在微芯片上进行表面等离子体共振成像，以高灵敏度检测沉积在表面上的非交联构型的dna连接金纳米颗粒。基于亲和电泳技术，在微流控通道中实现了多重基因突变分析。利用这一研究基金，我们通过设计各种dna共轭材料，成功地构建了生物传感系统。

We developed novel biosensing technologies on the basis of our unique principle, and presented facile, effective and high-sensitive analytical methods. The principle consists in single-base discrimination by using non-cross-linking aggregation of DNA-carrying nanoparticles, which are formed through the self-assembly of thermoresponsive polymers grafted with oligodeoxyribonucleotides. In this study, we devised new DNA-conjugated materials, including DNA-responsive hydrogels, DNA-synthetic polymer block-type conjugates, and DNA-linked gold nanoparticles, as analytical tools for gene diagnostics based on this principle.Achievements are summarized as follows:1. The unique aggregation of DNA-immobilized polystyrene Latex and DNA-linked gold nanoparticles was demonstrated, indicating that the principle would hold in general.2. Sequence-dependent, drastic change was found in electrophoretic mobility of DNA-carrying nanoparticles.3. Novel hybrid hydrogels containing rationally designed single-stranded DNA as the cross-linker were developed to shrink or swell in response to sequence of sample DNA, allowing for precise discrimination of single base difference.4. Affinity capillary electrophoresis using DNA-synthetic polymer block-type conjugates as affinity ligands was developed to separate single-stranded sample DNA and its point mutant.5. Surface plasmon resonance imaging was conducted on a microchip for high-sensitive detection of DNA-linked gold nanoparticles deposited onto the surface in a non-cross-linking configuration.6. Multiplex gene mutation analysis was realized on the basis of affinity electrophoresis in micro fluidic channels. With the aid of this research fund, we succeeded to construct biosensing systems by designing various DNA-conjugated materials.