A Mass Spectrometry Probe for Transient Imaging of In-Solution Biochemistry

描述(由申请人提供)：在生物医学研究中，经常需要检测和监测界面附近溶液中的动态化学成分。例如，许多研究的重点是检测培养组织或细胞向周围介质中的化学分泌物。通常，这样的应用对检测方法所需的空间和时间分辨率提出了特定的要求。在监测分泌物时，这是由于细胞类型和行为的异质性，以及细胞活动随时间的变化。例如，用荧光标记物、放射性标记物或使用抗原/抗体结合标记，作为成像动态生物化学的基础已经取得了惊人的成功，但人们对标记的分析物行为和非特异性结合的改变的担忧无法消除。此外，所有有针对性的方法，包括那些基于标记的方法，都固有地限制了它们的发现潜力，因为人们找不到自己不寻找的东西。这项研究的目的是克服目前生物化学成像技术的固有局限性。这将通过开发电喷雾离子源来实现，这种离子源可用作质谱学探针(MSP)，用于从生物界面附近的微环境进行高分辨率的生化检测。该研究团队已经证明了发明新型质谱学离子源的成功历史，并为这一项目提出了完成将样品收集、处理和电离的所有先决条件能力结合到微型采样毛细管中的雄心勃勃的任务。这种“尖端实验室”将包括在线微透析，以去除盐和交换溶剂，以及一个集成的胰酶消化微反应器。研究团队将通过建立一个多方面的方法来开发、优化和展示MSP，该方法结合了实验(包括光学和质量谱表征)、分析和模拟(第一原理物理模型和计算流体力学)和最先进的制造(微制造)。MSP将作为假说生成器在生物学研究中发挥重要作用，并将成为促进再生医学应用的生物反应器开发的关键工具。成功的结果有可能为广泛的研究应用带来变革的好处，包括生物标记物的发现，对健康和疾病细胞生物学的更好理解，生物传感器的开发，以及生物制造过程的分析和控制。除了在会议上作介绍和在档案期刊上发表文章外，还将通过在GA举办的教育讲习班传播MSP技术在生物学问题上的应用。技术部。此外，该探头将连接到TOF质谱仪，该质谱仪是NSF支持的国家纳米技术基础设施网络(NNIN)的一部分，因此可供来自工业和学术机构的用户使用。

DESCRIPTION (provided by applicant): Quite frequently in biomedical research there is a need to detect and monitor dynamic chemistries in solution in the vicinity of an interface. For instance, many studies focus on detection of chemical secretions from cultured tissues or cells into the surrounding medium. Often, such applications place specific demands on the required spatial and temporal resolution of the detection method. When monitoring secretions this would be due to heterogeneity in the cell types and behavior, and also variation in cellular activities with time. Labeling, with, for instance, a fluorescent marker, a radioactive marker, or using antigen/ antibody attachment, has been spectacularly successful as the foundation for imaging dynamic biochemistry, but concerns about the altering of labeled analyte behavior and non-specific binding cannot be eliminated. Furthermore, all targeted methods, including those based on labeling, are inherently limited in their discovery potential, as one cannot find what one is not looking for. The purpose of the proposed research is to overcome the inherent limitations of current biochemical imaging technologies. This will be done through the development of electrospray ion sources that can serve as mass spectrometry probes (MSP) for highly resolved biochemical detection from the microenvironment adjacent to biological interfaces. The research team has a demonstrated history of success inventing novel mass spectrometry ion sources, and proposes, for this project, to accomplish the ambitious task of combining all prerequisite capabilities for sample collection, processing, and ionization into a micro- sampling capillary. This "lab-on-a-tip" will include in-line microdialysis to remove salts and exchange solvent, as well as an integrated tryptic digestion micro-reactor. The research team will develop, optimize and demonstrate MSP through an established multifaceted approach combining experiment (including optical and mass spec characterization), analysis and simulation (first principles physical models and computational fluid dynamics), and state of the art manufacturing (microfabrication). MSP will assume an important role in biological research as a hypothesis generator, and will become a key tool in improving development of bioreactors for regenerative medicine applications. Successful results have potential for transformational benefits to a wide range of research applications, including biomarker discovery, improved understanding of healthy and diseased cell biology, biosensor development, and bio-manufacturing process analysis and control. In addition to presentation at conferences and publication in archival journals, the application of MSP technology to biological problems will be disseminated through an educational workshop hosted at Ga. Tech. Furthermore, the probe will be coupled to a TOF mass spectrometer that is part of the NSF supported National Nanotechnology Infrastructure Network (NNIN), and therefore available to users from industry and academic institutions alike.