Exploring Acoustic Disruption of Lipid Assemblies for Mass Spectrometry

探索用于质谱分析的脂质组装的声学破坏

• 批准号: 2108795
• 负责人: Theresa Evans-Nguyen
• 金额: $ 45.38万
• 依托单位: University of South Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2108795&HistoricalAwards=false
• 关键词: Exploring; Acoustic; Disruption; Lipid; Assemblies

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Theresa Evans-Nguyen and Venkat Bhethanabotla and their groups at the University of South Florida are devising ways to improve analysis of lipid assemblies critical to extracellular signaling. The roles of discrete lipids in these assemblies are not well understood. Furthermore, because lipids aggregate so readily, it is difficult even to identify and quantitate the individual lipid constituents of such assemblies. Mass spectrometry (MS) offers a powerful analytical tool which could provide a great deal of useful information if aggregation challenges could be overcome. To that end, this project focuses on using controlled acoustic waves to disrupt lipid aggregation, thereby facilitating MS analysis. The principles developed in this project should be extendable to other non-specific assemblies and amenable to coupling with current MS sampling methodologies. The research involves a cross-disciplinary effort between the fields of chemistry and engineering, and is expected to strengthen STEM recruitment programs through both fields.At the heart of this research is the use of novel surface acoustic wave (SAW) technologies in the MS workflow. In one modality, SAWs can already lyse extracellular vesicles known as exosomes. Also, as has been previously demonstrated by the team, one can use SAWs to mitigate biofouling from non-specific binding on microfluidic sensors. By extension, SAWs may be feasibly tuned to not just disrupt but further disintegrate bilayer membranes as model aggregates. In a second modality, SAW nebulization (SAWN) exhibits a distinct ionization capacity for both polar and non-polar lipids. By coupling to corona discharge (CD), the South Florida team has achieved comparable efficiency to electrospray ionization (ESI). Thus, the employment of SAWN-CD for the simultaneous analysis of non-polar cholesterol and polar phospholipids comprises a specific capability for which ESI falls short. SAWs are known to effect disruption of extracellular vesicles and to mitigate non-specific biofouling. In this work, SAWs are used to disintegrate model bilayer membrane vesicles into individual lipid components and, through SAW nebulization (SAWN), to facilitate MS ionization of both polar and non-polar lipids by coupling to corona discharge (CD). Specific objectives are: (1) formulation of liposomes as model aggregations to enable a systematic evaluation of SAWs; (2) investigation of SAW disruption parameters to disintegrate lipid assemblies; and 3) investigation SAW nebulization parameters to achieve simultaneous ionization of polar and non-polar species. Thus, through this investigation, the team aims to develop a method that can be applied to extracellular vesicles, allowing them to be directly sampled by mass spectrometry, vastly short-cutting traditional analysis methods for such complex assemblies.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学部化学测量和成像项目的支持下，南佛罗里达大学的Theresa Evans-Nguyen和Venkat Bhethanabotla及其团队正在设计方法来改善对细胞外信号至关重要的脂质组装的分析。离散脂质在这些组件中的作用还不清楚。此外，由于脂质如此容易聚集，甚至难以鉴定和定量这种组装体的单个脂质成分。质谱（MS）提供了一个强大的分析工具，如果可以克服聚集挑战，它可以提供大量有用的信息。 为此，该项目的重点是使用受控声波来破坏脂质聚集，从而促进MS分析。在这个项目中开发的原则应可扩展到其他非特定的组件和服从耦合与当前MS采样方法。 该研究涉及化学和工程领域之间的跨学科努力，预计将通过这两个领域加强STEM招聘计划。这项研究的核心是在MS工作流程中使用新型表面声波（SAW）技术。在一种模式中，SAW已经可以裂解被称为外泌体的细胞外囊泡。此外，正如该团队先前所证明的那样，人们可以使用SAW来减轻微流体传感器上非特异性结合的生物污染。通过扩展，SAW可以被可行地调整为不仅破坏而且进一步分解作为模型聚集体的双层膜。在第二种方式中，SAW雾化（SAWN）对极性和非极性脂质都表现出不同的电离能力。通过与电晕放电（CD）耦合，南佛罗里达团队已经实现了与电喷雾电离（ESI）相当的效率。因此，SAWN-CD用于同时分析非极性胆固醇和极性磷脂包括ESI福尔斯不足的特定能力。已知SAW影响细胞外囊泡的破坏并减轻非特异性生物污染。在这项工作中，SAW被用来分解模型双层膜囊泡成单独的脂质成分，并通过SAW雾化（SAWN），以促进极性和非极性脂质的MS电离耦合到电晕放电（CD）。具体目标是：（1）将脂质体配制为模型聚集体，以能够系统地评价SAW;（2）研究SAW破坏参数，以分解脂质组装体;以及3）研究SAW雾化参数，以实现极性和非极性物质的同时电离。因此，通过此次研究，该团队旨在开发一种可应用于细胞外囊泡的方法，使其能够通过质谱法直接采样，大大缩短了对这种复杂组装体的传统分析方法。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

项目成果

Combining plasmon-enhanced fluorescence with Rayleigh surface acoustic waves to quantify Carcinoembryonic Antigen from human plasma

结合等离子体增强荧光与瑞利表面声波来量化人血浆中的癌胚抗原

• DOI: 10.1016/j.bios.2022.114822
• 发表时间: 2023
• 期刊: Biosensors and Bioelectronics
• 影响因子: 12.6
• 作者: Huang, Yuqi;Li, Shuangming;Bhethanabotla, Venkat
• 通讯作者: Bhethanabotla, Venkat

Shake It Off! Acoustic Manipulation of Lipid Vesicles for Mass Spectrometric Analysis

• DOI: 10.1021/acs.analchem.3c01722
• 发表时间: 2023-08-30
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Taylor，Ashton N.;Huang，Yuqi;Evans-Nguyen，Theresa
• 通讯作者: Evans-Nguyen，Theresa

Orthogonal Surface Acoustic Wave (SAW) Sensor for Cancer Biomarker Detection with Accelerated Binding Kinetics

• DOI: 10.1109/sensors52175.2022.9967354
• 发表时间: 2022-10
• 期刊: 2022 IEEE Sensors
• 作者: Yuqi Huang;Maëlys Boucher;T. Evans-Nguyen;V. Bhethanabotla

Development of a thermo-pressure acoustic model and its application in modeling three-dimensional acoustofluidic systems

热压声学模型的开发及其在三维声流系统建模中的应用

• DOI: 10.1063/5.0140656
• 发表时间: 2023
• 期刊: Physics of Fluids
• 影响因子: 4.6
• 作者: Das, Pradipta Kr.;Bhethanabotla, Venkat R.
• 通讯作者: Bhethanabotla, Venkat R.