Nanofluidic Tools for Proteomic Sample Preparation

用于蛋白质组样品制备的纳米流体工具

• 批准号: 7121548
• 负责人: JONGYOON HAN
• 金额: $ 30.13万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7121548
• 关键词: biotechnology; microfluidics; nanotechnology; protein quantitation /detection; proteomics

Detailed proteomic analysis of biomolecule samples such as serum or cell extract could provide valuable information regarding the person's health or biological cells under study. However, current sample preparation technologies cannot deal with the diversity and dynamic range of these biomolecule samples effectively, and therefore the detection of information-rich minority protein species (signaling molecules and cytokines) is severely limited. Efficient protein separation in microfluidic systems could provide a solution for this challenge, but the limitation in sample volume, as well as the lack of molecular signal amplification strategy, has been major issues. Recent advances in nanofluidics provide useful tools for proteomic sample preparation, to address these challenges. Nanofluidic filters could be used for a gel-free separation of proteins based on the size. Permselective ion current generated by the nanofluidic channel was used to concentrate proteins and peptides from a dilute solution, to increase the sample volume capacity of the microfluidic sample preparation system. Also, continuous-flow pI-based biomolecule sorting system was demonstrated. In this project, we will develop these nanofluidic components further, and integrate them into an automatic proteomic sample preparation system that could sort out complex biomolecule sample based on the property of the target molecule. The flexibility and robustness of the micro/nanofluidic sample preparation devices that will be developed from this project will facilitate the system integration greatly. The resulting sample preparation microdevices could be easily integrated with various types of detectors, including antibody-based biosensor or mass spectrometry. Such an integration will enhance the selectivity and sensitivity of the biodetection by orders of magnitudes, therefore impacting various fields such as human health monitoring, biomarker detection, and systems biology.

对血清或细胞提取物等生物分子样品进行详细的蛋白质组学分析可以提供有关人体健康或所研究的生物细胞的有价值的信息。然而，当前的样品制备技术无法有效处理这些生物分子样品的多样性和动态范围，因此对信息丰富的少数蛋白质种类（信号分子和细胞因子）的检测受到严重限制。微流体系统中的高效蛋白质分离可以为这一挑战提供解决方案，但样品体积的限制以及分子信号放大策略的缺乏一直是主要问题。纳米流体学的最新进展为蛋白质组样品制备提供了有用的工具，以应对这些挑战。纳流体过滤器可用于根据尺寸进行蛋白质的无凝胶分离。纳流通道产生的选择性渗透离子流用于浓缩稀溶液中的蛋白质和肽，以增加微流控样品制备系统的样品容量。此外，还演示了基于连续流 pI 的生物分子分选系统。在这个项目中，我们将进一步开发这些纳米流体组件，并将它们集成到自动蛋白质组样品制备系统中，该系统可以根据目标分子的性质分选复杂的生物分子样品。该项目将开发的微纳流体样品制备装置的灵活性和鲁棒性将极大地促进系统集成。由此产生的样品制备微型设备可以轻松地与各种类型的检测器集成，包括基于抗体的生物传感器或质谱。这种集成将使生物检测的选择性和灵敏度提高几个数量级，从而影响人类健康监测、生物标志物检测和系统生物学等各个领域。