Plug-and-Play, Sensor-based, NanoLC/ESI Technology for Proteomics and Glycomics

用于蛋白质组学和糖组学的即插即用、基于传感器的 NanoLC/ESI 技术

• 批准号: 8057083
• 负责人: Colleen K Van Pelt
• 金额: $ 40万
• 依托单位: CORSOLUTIONS, LLC
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8057083
• 关键词: Address; Area; Back; Biological; Biological Sciences; Chemicals; Collaborations; Communities; Complex; Complex Mixtures; Computer software; Coupled; Custom; Data; Databases; Development; Device Designs; Devices; Diagnosis; Diagnostic; Dimensions; Docking; Electronics; Enhancement Technology; Gases; Generations; Genomics; Goals; Heating; Intervention; Laboratories; Laboratory Study; Length; Letters; Licensing; Liquid Chromatography; Marketing; Mass Spectrum Analysis; Monitor; Performance; Phase; Play; Plumbing; Polysaccharides; Post-Translational Protein Processing; Process; Proteins; Proteomics; Pump; Reading; Reproducibility; Resolution; Sample Size; Sampling; Scientist; Site; Small Business Innovation Research Grant; Solutions; Standardization; System; Techniques; Technology; Variant; Weight; Work; base; commercialization; complex biological systems; falls; improved; liquid chromatography mass spectrometry; mass spectrometer; nano; nano-electrospray; nanofluidic; novel; pressure; prevent; prototype; repaired; seal; sensor; tool; user-friendly; wound

DESCRIPTION (provided by applicant): Nano-liquid chromatography is a desirable technique because when coupled with mass spectrometry it can identify many of the components of a very complex proteomic or glycomic sample including subtle chemical variations within the sample, and can also obtain quantitative information. The technique is renowned for its high chromatographic resolution, high sensitivity, and small sample size requirement. But the technique remains cumbersome as it suffers from several limitations including system reproducibility issues created by user inabilities in column and/or spray emitter set-up, difficulty making nanofluidic connections, and difficulty troubleshooting system malfunctions. Others have tried to simplify this underutilized technique through primarily chip-based approaches, but have unfortunately compromised the technique's resolution and sensitivity. However, in a Phase I project, CorSolutions developed a robust, reliable, user-friendly nanoLC/ESI device that addresses these limitations of conventional nanoLC/ESI systems while still maintaining state-of- the-art performance, and is able to operate at elevated UHPLC pressures. These Phase I prototypes are the first plug-and-play nanoLC/ESI devices which do not compromise the high chromatographic resolution and sensitivity for which nanoLC is renowned. But although the technology developed in Phase I offers great improvements over conventional approaches, further developments are required for the technology to reach its large commercialization potential. The goals of the Phase II are to build upon the compression connection, plug-and-play technology developed in Phase I, in order to ready the technology for commercialization. This includes development of a cartridge-based insert for the column, allowing a column of any length to be wound in a small footprint. This enhancement will allow the overall device dimension to be substantially reduced. Additionally an emitter changer will be added, permitting the device to quickly exchange an emitter, in less than 1 sec, without need for user intervention. A spray sensor and leak sensors will be incorporated into the device with "smart" software to continuously monitor the sensor read back. The software will be capable of diagnosing system malfunctions and attempting automated repair such as an emitter change, greatly simplifying the troubleshooting process. Furthermore the ability to deliver nebulization gas will be incorporated into the device. These enhancements will allow for product commercialization. We see this simple, intuitive product becoming an integral part of biological laboratories, allowing standardization of proteomic and glycomic analyses. And most importantly the product will allow biologists to benefit from the highly informative nanoLC/MS data, enabling scientific breakthroughs and greater understanding of complex biological systems. This significant reduction in complexity of nanoLC will also allow for the technique to infiltrate deeper into the life sciences. PUBLIC HEALTH RELEVANCE: Nano-liquid chromatography mass spectrometry (nanoLC/MS) is a powerful technique for proteomic and glycomic studies which is able to not only identify proteins and glycans from very complex mixtures, but is also able to structurally characterize differences between the components, which is particularly important for post-translational modifications. However, conventional nanoLC/MS is a very difficult technique to perform, requiring a highly skilled user. Therefore this powerful technique, which could enable scientific discoveries and greater understanding of complex biological systems, is alarmingly underutilized. CorSolutions proposes to develop an intuitive, robust, reliable, user-friendly, "plug-and-play" device, which will address the challenges of conventional nanoLC/MS, dramatically reducing the complexity of the technique and thereby allowing its pervasive use in biological laboratories.

描述(申请人提供)：纳米液相色谱是一种理想的技术，因为当与质谱学结合时，它可以识别非常复杂的蛋白质组或糖类样品的许多成分，包括样品中细微的化学变化，还可以获得定量信息。该技术以其高层析分辨率、高灵敏度和小样本量要求而闻名。但这项技术仍然很繁琐，因为它受到几个限制，包括由于柱和/或喷雾发射器设置中的用户能力不佳而造成的系统重复性问题，难以进行纳米流体连接，以及难以排除系统故障。其他人试图通过主要基于芯片的方法来简化这种未得到充分利用的技术，但不幸的是，这项技术的分辨率和灵敏度都受到了影响。然而，在第一阶段项目中，CorSolutions开发了一种坚固、可靠、用户友好的NanoLC/ESI设备，该设备解决了传统NanoLC/ESI系统的这些局限性，同时仍保持最先进的性能，并能够在较高的UHPLC压力下运行。这些第一阶段的原型是第一批即插即用的NanoLC/ESI设备，不会影响NanoLC闻名于世的高层析分辨率和灵敏度。但是，尽管在第一阶段开发的技术比传统方法有了很大的改进，但要使该技术发挥其巨大的商业潜力，还需要进一步的开发。第二阶段的目标是在第一阶段开发的即插即用压缩连接技术的基础上，为该技术的商业化做好准备。这包括开发用于柱的基于筒的插入件，从而允许将任意长度的柱缠绕在较小的占地面积中。这一改进将允许显著减小器件的整体尺寸。此外，还将添加发射器转换器，允许设备在不到1秒的时间内快速更换发射器，无需用户干预。喷雾传感器和泄漏传感器将被集成到该设备中，并带有“智能”软件，以持续监测传感器读出的情况。该软件将能够诊断系统故障并尝试自动修复，例如更换发射器，极大地简化故障排除过程。此外，该设备还将具备输送雾化气体的能力。这些改进将使产品商业化。我们看到这种简单、直观的产品成为生物实验室不可或缺的一部分，允许蛋白质组和血糖分析的标准化。最重要的是，该产品将使生物学家受益于高度信息量的NanoLC/MS数据，使科学突破和对复杂生物系统的更好理解成为可能。纳米LC复杂性的显著降低也将使这项技术能够更深入地渗透到生命科学中。 公共卫生相关性：纳米液相色谱质谱(NanoLC/MS)是蛋白质组和糖学研究的一种强大技术，它不仅能够从非常复杂的混合物中识别蛋白质和多糖，而且能够从结构上表征组分之间的差异，这对翻译后修饰尤为重要。然而，传统的NanoLC/MS是一项非常困难的技术，需要高度熟练的用户。因此，这项强大的技术，可以使科学发现和更好地了解复杂的生物系统，但令人震惊的是没有得到充分利用。CorSolutions建议开发一种直观、坚固、可靠、用户友好的“即插即用”设备，它将解决传统NanoLC/MS的挑战，极大地降低技术的复杂性，从而使其能够在生物实验室中广泛使用。