NER: Signal Amplification in Biomolecular Detectors Using Activated Receptor Molecules

NER：使用激活受体分子的生物分子检测器信号放大

• 批准号: 0508398
• 负责人: Cagri Savran
• 金额: $ 10万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0508398&HistoricalAwards=false
• 关键词: NER; Signal; Amplification; Biomolecular; Detectors

The objective of this research is to increase the sensitivity of label-free biomolecular detectors to levels achieved by label-based detectors (e.g. fluorescence) that are sensitive but time-consuming and costly. A method that improves sensitivity without modification of sensor structure or operation principle is highly desirable. The approach is to incorporate activated receptor molecules into biosensors. Single-stranded DNA or RNA-based molecules, namely aptazymes that perform enzyme activity upon binding to small targets will be used as receptor molecules. Accordingly, biosensors will detect the activity of relatively large aptazymes as opposed to directly detecting the binding of small target molecules. Since the activity will be triggered by small target molecules, they will be indirectly detected with greatly amplified sensitivity. The efficacy of the technique will be demonstrated on the surfaces of a few model label-free detection systems (both commercial and nanomechanical sensors built in-house) whereby small molecules whose direct binding would reveal signals too small to detect will be indirectly detected via aptazyme activity. All experimental work will be undertaken by Purdue. The aptazyme synthesis will be performed at the University of Texas at Austin. This research project intends to make a broad impact on high-throughput detection of biomolecules such as DNA, RNA, proteins and small molecules whose sensitive detection is of top importance for early diagnosis of many cancers that take 500,000 American lives every year. The findings of this project will be a significant step towards the development of fast, low-cost and high-resolution biomolecular detectors and towards the understanding of biomolecular interactions on solid surfaces. This project will also have significant educational contributions by setting an interdisciplinary stage where engineering students and chemists can interact and learn from each other. A part of the project will also be linked to a graduate level laboratory course.

这项研究的目的是提高无标记生物分子检测器的灵敏度，使其达到基于标记的检测器（例如荧光）所达到的水平，这些检测器灵敏但耗时且昂贵。非常需要一种在不修改传感器结构或操作原理的情况下提高灵敏度的方法。该方法是将激活的受体分子纳入生物传感器。基于单链DNA或RNA的分子，即在与小靶标结合时执行酶活性的适体酶将用作受体分子。因此，生物传感器将检测相对大的适体酶的活性，而不是直接检测小靶分子的结合。由于活性将由小的靶分子触发，因此它们将以极大放大的灵敏度被间接检测。该技术的有效性将在几个模型的无标记检测系统（商业和内部构建的纳米机械传感器）的表面上得到证明，其中直接结合会显示信号太小而无法检测的小分子将通过aptazyme活性间接检测。所有实验工作将由普渡大学承担。适体酶的合成将在德克萨斯大学奥斯汀分校进行。该研究项目旨在对DNA、RNA、蛋白质和小分子等生物分子的高通量检测产生广泛影响，这些生物分子的灵敏检测对于每年夺走50万美国人生命的许多癌症的早期诊断至关重要。该项目的研究结果将是朝着开发快速，低成本和高分辨率生物分子探测器以及了解固体表面上生物分子相互作用的重要一步。该项目还将通过建立一个跨学科的阶段，使工程专业的学生和化学家可以相互交流和学习，从而做出重大的教育贡献。该项目的一部分还将与研究生水平的实验室课程相联系。