Improved Methods and Commercialization of Native Mass Spectrometry by Capillary Vibrating Sharp-edge Spray Ionization (cVSSI)

毛细管振动锐边喷雾电离 (cVSSI) 天然质谱的改进方法和商业化

• 批准号: 10256885
• 负责人: Stephen Valentine
• 金额: $ 25.65万
• 依托单位: INVIBRAGEN INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256885
• 关键词: 3-Dimensional; Adoption; Affect; Amplifiers; Area; Benchmarking; Biological Response Modifier Therapy; Biopolymers; Blood capillaries; Buffers; Clip; Communicable Diseases; Coupled; Coupling; Data; Deposition; Detection; Deuterium; Development; Devices; Disease; Disease Progression; Exhibits; Future; Gases; Goals; Growth; Health; Housing; Human; Hydrogen; Industry; Infusion procedures; Ions; Isoelectric Point; Letters; Liquid Chromatography; Malignant Neoplasms; Manufacturer Name; Mass Spectrum Analysis; Medicine; Methods; Micromanipulation; Nebulizer; Oligonucleotides; Onset of illness; Pathway interactions; Performance; Pharmacologic Substance; Phase; Positioning Attribute; Power Sources; Production; Proliferating; Protein Analysis; Protein Engineering; Proteins; Research; Research Personnel; Signal Transduction; Solvents; Source; Structure; System; Technology; Therapeutic; Universities; West Virginia; Work; aqueous; base; commercialization; design; design and construction; early onset; experimental study; improved; instrument; instrumentation; interest; ion source; ionization; ionization technique; mass analyzer; mass spectrometer; nano-electrospray; new technology; novel strategies; preservation; protein complex; protein structure; prototype; therapeutic gene; therapeutic protein; three dimensional structure; tool; voltage

Abstract/Summary Over the last 20 years, mass spectrometry (MS) instrumentation has advanced at a remarkable clip. Improvements in ion focusing, mass analyzer, and ion detection regions have greatly enabled the study of protein molecules in their native state (native MS). One area that has remained relatively constant is the ionization source where nanoelectrospray ionization (nESI) has remained the most sensitive means to produce the large biomolecules of interest. A severe drawback of nESI for native MS is that it suffers from stability issues when operated in negative ion mode. This primarily results from the earlier onset of the corona discharge in negative ion mode which abrogates ion production. The result of this limitation is to drastically reduce the utility of native MS as >104 proteins exhibit PI values below 6. This has implications for the study of protein structures important to human health ranging from therapeutic molecules to those associated with disease onset and progression. Recently, the research group of Prof. Peng Li at West Virginia University developed a new ionization technique termed Vibrating Sharp-edge Spray Ionization (VSSI). The approach has recently been demonstrated to provide a 10× to 100× boost in ionization performance compared with state-of-the-art approaches operated in negative ion mode. Here, we will leverage the power of the new approach to provide a prototype ionization source which will consistently provide 100× signal enhancements in negative ion mode and 10× improvement in positive ion mode. These leaps in ionization efficiency will unlock the door to high-efficiency native MS experiments especially in negative ion mode. The result will be to enable human health researchers to characterize the structures of many important proteins that have heretofore remained largely inaccessible.

摘要/概要 在过去的20年里，质谱（MS）仪器以惊人的速度发展。 剪辑.离子聚焦、质量分析器和离子检测区域中的改进极大地改善了离子聚焦、质量分析器和离子检测区域。 使研究蛋白质分子在其天然状态（天然MS）。的一个领域 保持相对恒定的是电离源，其中纳米电喷雾电离（nESI） 仍然是生产感兴趣的大生物分子的最灵敏的方法。严重 用于天然MS的nESI的缺点是当在负离子环境下操作时， 离子模式。这主要是由于在负离子中电晕放电开始较早所致 消除离子产生的模式。这种限制的结果是大大减少了 天然MS的效用，因为>104蛋白质表现出低于6的PI值。这影响到 研究对人类健康重要的蛋白质结构，从治疗分子到 与疾病发作和进展相关的。最近，教授的研究小组。 西弗吉尼亚大学的李鹏开发了一种名为振动的新电离技术 锐边喷雾电离（VSSI）。该方法最近被证明可以提供 与最先进的方法相比，电离性能提高10倍至100倍 以负离子模式运行。在这里，我们将利用新方法的力量， 提供一个原型电离源，可持续提供100×信号增强 在负离子模式下性能提高10倍，在正离子模式下性能提高10倍。电离的这些跳跃 效率将打开大门，以高效率的本地MS实验，特别是在负 离子模式。其结果将使人类健康研究人员能够表征这些结构 许多重要的蛋白质，迄今为止仍然很大程度上无法获得。