基于刺激响应型纳米容器探针的无标记高灵敏高通量生物传感研究

Stimuli-responsive nanomaterial has unique nanostructure and fascinating properties, such as controllable three-dimensional structure, good biocompatibility and mechanical properties, and responding to the external environment stimulation. The aim of this study is to synthesis stimuli-responsive nanomaterial (such as mesoporous silica, mesoporous carbon, etc.) for label-free, high sensitivity, high throughput and visual determination of miRNA by the aid of electrochemical, optical detection method and microchip technology. We intend to use stimuli-responsive nanomaterial as nano-reservoir to encapsulate a lot of signal substances and to immobilize probe molecule on the surface, developing novel method for label-free and high sensitivity determination of miRNA; Also, we want to couple the amplification of stimuli-responsive nano-reservoir with the biotechnology (such as enzyme catalyzed amplification, rolling circle amplification, DNA self-assembly etc.) to further enhance the detection signal; At the same time, we plan to immobilize different probe molecules on different nano-reservoirs surface which encapsulated different signal substances for high sensitive and simultaneous determination of multiple miRNAs; In addition, we plan to further study the characteristics of stimuli-responsive nanomaterial, explore the trigger mechanism of various stimuli-responsive, construct the novel label-free analysis strategies and methods, realize the simultaneous detection of many kind of biomarkers (such as protein, hormone etc.). Finally, we try to combine these novel methods with microchip, development of novel, low cost, rapid, sensitive, portable biomarker detection device.

刺激响应型纳米材料具有可控的三维结构，良好的生物相容性和机械性能，且能对外界环境刺激做出响应等优点。本项目拟合成刺激响应型纳米材料（如介孔二氧化硅、介孔碳等），借助电化学、光化学检测技术并辅以微芯片实现对miRNA无标记的高灵敏高通量及可视化检测。将刺激响应型纳米材料作为纳米容器封装大量信号物质同时在其表面固定探针分子，实现对miRNA无标记的高灵敏检测；将刺激响应型纳米容器放大同生物放大技术（如酶放大、滚环扩增放大、DNA自组装等）相联合，实现信号二次放大；将不同探针分子修饰在封装有不同信号物质的不同纳米容器表面，实现miRNA多目标同时高灵敏检测；同时，深入研究刺激响应型纳米材料的特性，探索不同刺激响应的触发机制，构建新型的无标记检测分析策略和方法，实现多类生物标志物（如蛋白、激素等）的同时检测。最后将该方法集成到微芯片上，构建低成本、快速、灵敏、便携的生物标志物检测新设备。

本项目研究采取多种合成方法制备了一系列新型的碳基（如介孔硅、石墨烯、碳点）纳米复合材料，借助电化学、光化学检测技术实现了miRNA无标记的高灵敏高通量检测。利用介孔硅封装电活性物质硫堇同时在介孔硅表面固定探针分子，实现了miRNA-141无标记的高灵敏检测；将介孔硅作为纳米容器封装CdS和PbS量子点，同时在其表面固定相应的探针分子，实现了乳腺癌相关miRNA（miRNA-21和miRNA-205）的同时高灵敏无标记检测。另外，我们还将构建的电化学、光电化学、比色和荧光传感器应用于生物分子、药物分子、环境污染物、重金属离子的高灵敏检测中，在此基础上均取得了良好的研究成果。