NanoArrays in Mass Spectrometry for Proteomic and Metabolomic Applications

描述（由申请人提供）：我们的长期目标是开发纳米阵列作为蛋白质组学和代谢组学应用的高特异性，高灵敏度和超高通量的质谱平台。这将建立在两个具有互补专业知识的实验室的密切合作之上：Wang （LBNL）在质谱和蛋白质组学方面，Yang （LBNL/加州大学伯克利分校）在纳米科学和纳米技术方面。蛋白质组学和代谢组学作为系统生物学的组成部分，将为假设驱动的生化和生物成像研究提供新的知识库。需要进一步发展使能技术-质谱-以显着提高灵敏度和动态范围。最终实现单细胞的蛋白质组学和代谢组学需要革命性的创新。纳米结构被定义为在纳米尺度上至少具有一维的结构，并且由于其迷人的特性和优于其块状对应物的独特应用而引起了广泛的兴趣。通过制造新型纳米结构或简单地将现有微结构的尺寸缩小到纳米级，存在令人兴奋的机会。在这个探索性/发展性R21项目中，我们建议开发新的纳米阵列基底物用于质谱分析，并展示其在蛋白质组学和代谢组学中的应用。具体来说，我们建议开发一种新的电离源。我们将采取多方面的方法来制造和表征这些设备（目标1），阐明电离机制（目标2），并展示在质谱分析中对单个干细胞蛋白质组学的原理验证应用（目标3）。我们在纳米线和纳米管阵列的制造以及纳米器件和微器件的集成方面的广泛能力和专业知识，将使我们处于一个独特的位置，以实现潜在的突破。如果成功，这些纳米器件将成为纳米流体和微流体结构中分离、检测和识别生物分子的关键组成部分，并为系统生物学开辟新的可能性。与公共卫生相关：干细胞可以自我更新，在再生医学中显示出巨大的希望。最终阐明单细胞水平的干细胞分化需要革命性的创新。为了实现这一目标，我们建议利用纳米技术的巨大潜力来开发新的电离底物，用于质谱研究单个干细胞的蛋白质和代谢物。

DESCRIPTION (provided by applicant): Our long term goal of this program is to develop nanoarrays as highly-specific, highly-sensitive and ultrahigh-throughput platforms in mass spectrometry for proteomic and metabolomic applications. This will be built upon the close collaborations of 2 laboratories with complementary expertise: Wang (LBNL) in mass spectrometry and proteomics, and Yang (LBNL/UC Berkeley) in nanoscience and nanotechnology. Proteomics and metabolomics as integral systems biology components will generate a new knowledgebase for hypothesis-driven biochemical and bioimaging studies. Further developments in the enabling technology- mass spectrometry-are required to significantly increase the sensitivity and dynamic range. Revolutionary innovations are needed to ultimately achieve proteomics and metabolomics of single cells. Nanostructures are structures defined as having at least 1 dimension at nanometer scale, and have generated intensive interests as a result of their fascinating properties, and unique applications superior to their bulk counterparts. Exciting opportunities exist by making new types of nanostructures or simply by shrinking the size of existing microstructures into the nanometer regime. In this exploratory/ developmental R21 project, we propose to develop novel nanoarray-based substrates for mass spectrometry and demonstrate their applications in proteomics and metabolomics. Specifically we propose to develop a new kind of ionization source. We will take a multi-faceted approach to fabricate and characterize the devices (Aim 1), elucidate the ionization mechanisms (Aim 2), and demonstrate the proof-of-principle applications in mass spectrometry for single stem cell proteomics (Aim 3). Our extensive capabilities and expertise in the fabrication of nanowire and nanotube arrays, and in the integration of nanodevices and microdevices, will put us in a unique position to achieve potential breakthroughs. If successful, these nano devices will serve as a critical component in the nanofluidics and microfluidics structures to separate, detect, and identify biomolecules, and open up new possibilities in systems biology. Relevance to public health: Stem cells can self-renew and show great promise in regenerative medicine. Revolutionary innovations are needed to ultimately elucidate stem cell differentiation at single cell level. Towards this goal, we propose to utilize the enormous potential of nanotechnology to develop novel ionization substrates for mass spectrometry to study proteins and metabolites of single stem cells.