Suspended microchannels for biomolecular detection

用于生物分子检测的悬浮微通道

• 批准号: 6755848
• 负责人: SCOTT R MANALIS
• 金额: $ 22.23万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6755848
• 关键词: bioengineering /biomedical engineering; biomedical equipment development; fluid flow; physical chemical interaction; protein binding; protein quantitation /detection

DESCRIPTION (provided by applicant): Proteins and their byproducts play a critical role in nearly every event that takes place within living cells. The understanding of many of these events is furthered by the ability to profile the concentration of specific proteins as a function of time and various physiological conditions. However, the rate at which these parameters are measured by current methodology is often limited by requirements for large sample volumes and time-consuming modifications of the molecules under investigation. Here we address these limitations by proposing a detection system based on microfluidic channels whereby the channels themselves are the detectors. The key innovation is that the mechanical properties of a suspended microfluidic channel with a thickness near a micron will allow protein concentration to be determined by measuring a mass-dependent resonant frequency of the microchannel. The suspended channel mass is altered either by proteins that enter into the channel volume, or by specific binding of proteins to capture molecules on the channel walls. Successful development of the suspended microchannel detector will result in a real-time and labelfree method that will provide an alternative to fluorescence for protein detection. Specifically, suspended microchannel detectors will offer two advances. First, is scalability: ultimately 10[2]-10[3] detectors could be operated within a square centimeter with a volume of less than 100 pL per detector. Second, we estimate that the suspended microchannel will achieve a detection limit in the range of 10[-18] is to 10[-19] g/mu m 2. This limit, which corresponds to less than 10 proteins/mu m 2, is nearly 100-fold more sensitive than other label-free methods such as the quartz crystal microbalance. We anticipate that an array of suspended microchannel detectors will substantially increase the throughput and utility of protein assays that are used for furthering the predictive power of system-level modeling. The suspended microchannels will also be useful for enabling point-of-care medical diagnostics.

描述（由申请人提供）：蛋白质及其副产品在几乎所有发生在活细胞内的事件中都起着至关重要的作用。通过将特定蛋白质的浓度作为时间和各种生理条件的函数概述的能力来进一步了解这些事件的理解。但是，通过当前方法测量这些参数的速率通常受到对所研究分子的大量样品体积和耗时修改的要求的限制。在这里，我们通过基于微流体通道提出检测系统来解决这些限制，该系统本身就是检测器。关键创新是，悬浮的微流体通道的机械性能在微米附近厚度，将允许通过测量微通道的质量依赖性谐振频率来确定蛋白质浓度。悬浮的通道质量会被输入通道体积的蛋白质或蛋白质特异性结合以捕获通道壁上的分子而改变。悬浮的微通道检测器的成功开发将导致一种实时和Labelfree方法，该方法将为荧光提供蛋白质检测的替代方法。具体而言，悬挂的微通道检测器将提供两个进步。首先，是可伸缩性：最终可以在平方厘米内操作10 [2] -10 [3]，每个检测器的体积小于100 pl。其次，我们估计悬浮的微通道将达到10 [-18]范围内的检测极限，为10 [-19] g/mu m 2。该极限对应于小于10蛋白/mu m 2，比其他不含标签的方法（例如石英晶体微生物）要敏感了近100倍。我们预计，一系列悬浮的微通道检测器将大大增加用于促进系统级建模的预测能力的蛋白质测定的吞吐量和效用。悬浮的微通道也将用于实现护理点医学诊断。