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

用于溶液内生物化学瞬态成像的质谱探针

• 批准号: 8322652
• 负责人: ANDREI G FEDOROV
• 金额: $ 18.48万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322652
• 关键词: Adsorption; Affect; Antibodies; Antigens; Behavior; Binding; Biochemical; Biochemistry; Biological; Biological Markers; Biomedical Engineering; Biomedical Research; Bioreactors; Biosensor; Blood capillaries; Cells; Cellular biology; Chemicals; Chemistry; Complex; Coupled; Data; Detection; Development; Dialysis procedure; Digestion; Educational workshop; Electrodes; Environment; Enzymes; Event; Foundations; Gases; Generations; Goals; Guidelines; Heterogeneity; Image; Imaging technology; In Vitro; Industry; Infusion procedures; Institution; Investigation; Ion Transport; Journals; Kinetics; Label; Liquid substance; Location; Mass Spectrum Analysis; Measurement; Membrane; Methods; Microdialysis; Microfabrication; Modification; Monitor; Nanotechnology; Optics; Outcome; Physiological; Preparation; Process; Proteins; Publications; Radioactive; Recording of previous events; Regenerative Medicine; Research; Research Infrastructure; Resolution; Role; Salts; Sampling; Sodium Chloride; Solutions; Solvents; Technology; Time; Variant; base; biological research; capillary; cell behavior; cell type; electric field; improved; innovation; insight; ion source; ionization; manufacturing process; mass spectrometer; nanolitre; nanopattern; novel; novel strategies; physical model; pressure; research study; sample collection; simulation; success; symposium; tissue culture; tool

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.

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物质。例如，许多研究侧重于检测培养组织或细胞向周围介质中的化学分泌物。通常，此类应用对检测方法所需的空间和时间分辨率提出特定要求。当监测分泌物时，这可能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，使用荧光标记、放射性标记或使用抗原/抗体附着进行标记，作为动态生物化学成像的基础已经取得了巨大成功，但对标记分析物行为和非特异性结合的改变的担忧不能被消除。被淘汰。此外，所有有针对性的方法，包括基于标记的方法，其发现潜力本质上都受到限制，因为人们无法找到不需要的东西。拟议研究的目的是克服当前生化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，电喷雾离子源可以用作质谱探针（MSP），用于从生物界面附近的微环境中进行高分辨率的生化检测。该研究团队在发明新型质谱离子源方面有着成功的历史，并建议在该项目中完成将样品收集、处理和电离的所有先决条件结合到微采样毛细管中的雄心勃勃的任务。这个“尖端实验室”将包括用于去除盐和交换溶剂的在线微透析，以及集成的胰蛋白酶消化微反应器。研究团队将通过既定的多方面方法，结合实验（包括光学和质谱表征）、分析和模拟（第一原理物理模型和计算流体动力学）以及最先进的制造（微加工）来开发、优化和演示 MSP。 MSP 将作为假设生成器在生物学研究中发挥重要作用，并将成为改善再生医学应用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性的好处，包括生物标志物发现、提高对健康和患病细胞生物学的理解、生物传感器开发以及生物制造过程分析和控制。除了在会议上发表演讲和在档案期刊上发表文章外，MSP 技术在生物问题上的应用还将通过在 Ga.Tech 举办的教育研讨会进行传播。此外，该探针将与 TOF 质谱仪耦合，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，因此可供行业和学术机构的用户使用。