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: 7390437
• 负责人: Colleen K Van Pelt
• 金额: $ 20万
• 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 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7390437
• 关键词: Address; Area; Arts; Attention; Biological; Biological Sciences; Boxing; Chemicals; Chromatography; Complex; Complex Mixtures; Computer software; Coupled; Daily; Data; Data Analyses; Data Quality; Databases; Devices; Diagnosis; Diagnostic; Docking; Electronics; Feedback; Genomics; Goals; Laboratories; Laboratory Study; Legal patent; Liquid Chromatography; Mass Spectrum Analysis; Monitor; Numbers; Performance; Phase; Play; Plumbing; Polysaccharides; Post-Translational Protein Processing; Proteins; Proteomics; Pump; Reproducibility; Research Personnel; Sample Size; Sampling; Site; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Solvents; System; Techniques; Technology; Time; Today; Variant; Water; Work; base; design; desire; glycosylation; interest; ionization; liquid chromatography mass spectrometry; mass spectrometer; nano; nanofluidic; novel; prevent; prototype; seal; sensor; user-friendly

DESCRIPTION (provided by applicant): Conventional nano-liquid chromatography electrospray mass spectrometry (nanoLC/ESI/MS or nanoLC/MS) is a challenging technique to perform, requiring a highly proficient user, and consequently, this powerful technique is alarmingly underutilized. The difficulty in nanoLC/MS resides in the nanoLC and electrospray moieties, as mass spectrometers and database search software have become increasingly user friendly and intuitive. NanoLC electrospray systems suffer from several major limitations including: 1) system reproducibility issues created by user inabilities in column and/or sprayer set-up; 2) difficulty making nano-fluidic connections; 3) difficulty troubleshooting system malfunctions; 4) technique is very labor intensive, especially at start-up; 5) cannot optimize chromatography and ionization solvent independently within a single nanoLC/MS analysis. CorSolutions has designed and will develop a robust, reliable, and user friendly nano-liquid chromatography electrospray device that will address each of the limitations of conventional nanoLC electrospray systems while still maintaining state-of-the-art performance. The CorSolutions nanoLC electrospray technology consists of conventional nanoLC pumps and a nanoLC autosampler, a hardware docking unit which mounts directly in front of the mass spectrometer orifice, and a "pop-in" consumable device which inserts into the hardware docking unit. The "pop-in" insert is a sealed vessel which serves as a black-box to the user. In the device is imbedded a nanoLC column and spray emitter, as well as multiple sensors to diagnose and troubleshoot system issues. When the consumable device is inserted into the docking hardware, the notoriously difficult nanoLC plumbing connections are automatically made using novel "plug and play" technology. The system offers the user an easy, intuitive means of performing nanoLC and electrospray, without needing to be skilled in the art. The goals of the Phase I SBIR will be to first construct a prototype system. This will include designing and building "plug and play" compression fittings, an automated compression fitting connection mechanism, a "pop-in" insert containing a nanoLC column and sprayer, sensors to detect system leaks and monitor spray current, as well as sensor electronics. The second goal will be to evaluate the functionality of the prototype system. This will include comparing data quality between the prototype and a conventional nanoLC system, and placing the prototype in a proteomics laboratory for "real world" feedback. Our long term objective is to provide a robust, reliable nanoLC electrospray device, which can be coupled to a wide variety of mass spectrometers. We see this simple, intuitive product becoming an integral part of biological laboratories. 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.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 glycosylation 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 pervasive use of the technology in all biological laboratories.

描述（由申请人提供）：传统的纳米液相色谱电喷雾质谱（nanoLC/ESI/MS或nanoLC/MS）是一项具有挑战性的技术，需要高度熟练的用户，因此，这种强大的技术未得到充分利用。nanoLC/MS的困难在于nanoLC和电喷雾部分，因为质谱仪和数据库搜索软件变得越来越用户友好和直观。NanoLC电喷雾系统遭受几个主要限制，包括：1）由柱和/或喷雾器设置中的用户不稳定性产生的系统再现性问题; 2）难以进行纳米流体连接; 3）难以对系统故障进行故障排除; 4）技术是非常劳动密集的，特别是在启动时; 5）不能在单个nanoLC/MS分析中独立地优化色谱和电离溶剂。CorSolutions已经设计并将开发一种坚固、可靠和用户友好的纳米液相色谱电喷雾设备，该设备将解决传统纳米液相色谱电喷雾系统的每一个局限性，同时仍保持最先进的性能。CorSolutions nanoLC电喷雾技术由传统的nanoLC泵和nanoLC自动进样器、直接安装在质谱仪孔前的硬件对接单元以及插入硬件对接单元的“弹出式”耗材设备组成。“弹出”插入物是密封容器，其充当用户的黑盒。在该设备中嵌入了nanoLC柱和喷雾发射器，以及多个传感器来诊断和排除系统问题。当消耗性设备插入对接硬件时，使用新颖的“即插即用”技术自动进行众所周知的困难的nanoLC管道连接。该系统为用户提供了一种简单、直观的方法来执行nanoLC和电喷雾，而不需要本领域的技术人员。第I阶段SBIR的目标是首先构建原型系统。这将包括设计和建造“即插即用”压缩配件、自动压缩配件连接机制、包含nanoLC柱和喷雾器的“弹出”插件、检测系统泄漏和监测喷雾电流的传感器以及传感器电子器件。第二个目标是评价原型系统的功能。这将包括比较原型和传统nanoLC系统之间的数据质量，并将原型放置在蛋白质组学实验室中以获得“真实的世界”反馈。我们的长期目标是提供一个强大的，可靠的nanoLC电喷雾装置，它可以耦合到各种各样的质谱仪。我们看到这个简单、直观的产品成为生物实验室不可或缺的一部分。最重要的是，该产品将使生物学家能够从信息丰富的nanoLC/MS数据中受益，从而实现科学突破并更好地了解复杂的生物系统。（nanoLC/MS）是用于蛋白质组学和糖组学研究的强大技术，其不仅能够从非常复杂的混合物中鉴定蛋白质和聚糖，而且还能够在结构上表征组分之间的差异，这对于糖基化翻译后修饰特别重要。然而，传统的nanoLC/MS是一种非常难以执行的技术，需要高度熟练的用户。因此，这种强大的技术，可以使科学发现和更好地了解复杂的生物系统，是令人震惊的利用不足。CorSolutions提出开发一种直观、强大、可靠、用户友好的“即插即用”设备，该设备将解决传统nanoLC/MS的挑战，大大降低技术的复杂性，从而使该技术在所有生物实验室中得到普遍使用。