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）：蛋白质及其副产物在活细胞内发生的几乎所有事件中都发挥着关键作用。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进行分析的能力，进一步加深了对许多这些事件的理解。然而，当前方法测量这些参数的速率通常受到大样本量的要求和所研究分子的耗时修改的限制。在这里，我们通过提出一种基于微流体通道的检测系统来解决这些限制，其中通道本身就是检测器。关键的创新在于，厚度接近微米的悬浮微流体通道的机械特性将允许通过测量微通道的质量依赖共振频率来确定蛋白质浓度。悬浮的通道质量可以通过进入通道体积的蛋白质或通过蛋白质的特异性结合来改变以捕获通道壁上的分子。悬浮微通道检测器的成功开发将产生一种实时、无标记的方法，为蛋白质检测提供荧光的替代方法。具体来说，悬挂式微通道探测器将带来两大进步。首先是可扩展性：最终可以在一平方厘米内运行 10[2]-10[3] 个探测器，每个探测器的体积小于 100 pL。其次，我们估计悬浮微通道将达到 10[-18] 至 10[-19] g/mu m 2 范围内的检测极限。该极限对应于小于 10 个蛋白质/μ m 2，比石英晶体微天平等其他无标记方法的灵敏度高近 100 倍。我们预计，一系列悬浮微通道检测器将大大提高蛋白质检测的通量和实用性，用于进一步提高系统级建模的预测能力。悬浮的微通道也将有助于实现即时医疗诊断。