RUI: Development of Fluorescent Free-Radical Tags for N-Glycan Quantitation and Characterization using UPLC-MS/MS

RUI：使用 UPLC-MS/MS 开发用于 N-聚糖定量和表征的荧光自由基标签

• 批准号: 2107798
• 负责人: Jinshan Gao
• 金额: $ 31.5万
• 依托单位: Montclair State University
• 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: 2021-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2107798&HistoricalAwards=false
• 关键词: RUI; Development; Fluorescent; Free; Radical

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Jinshan Gao and his students at Montclair State University are pursuing research that seeks improved capabilities for simultaneous characterization and quantitation of an important class of biomolecules known as glycans – the carbohydrate-modifications that decorate so many proteins in biological systems and that are of great significance in biomedical science. Dr. Gao's approach uses mass spectrometry (MS), one of the most widely used and powerful tools of chemical analysis. Specifically, he is developing ways to label glycan molecules with fluorescent chemical "tags" to achieve absolute quantitation and to facilitate their structural characterization. The impact of proposed research is expected to be enhanced by the involvement of undergraduate students, including first-generation college students and members of underrepresented groups in science - Montclair State University is a Hispanic-Serving Institution and Primarily Undergraduate Institution. Elements of the research are integrated into Dr. Gao's undergraduate and graduate courses, enabling students to become familiar with modern analytical techniques and helping them to develop long-term interests for possible careers in the STEM (Science, Technology, Engineering and Mathematics) fields.The Gao group is developing novel fluorescent free-radical tags for simultaneous glycan quantitation and characterization, and novel multiplexed, isobaric fluorescent free-radical tags for simultaneous characterization and quantitation of each glycan from multiple samples. The tags contain four equally important chemical characteristics: 1) the fluorophore facilitates highly sensitive and reliable quantitation; 2) the free-radical precursor is expected to induce systematic, efficient, and predictable fragmentation for structure characterization; 3) a hydrazide or oxyamine group selectively targets the reducing terminus of the glycan; and 4) a pyridine or tertiary amine group is expected to significantly enhance the MS signal. These tailored tags are designed to enable systematic, efficient, and predictable free-radical-mediated glycan dissociation, allowing efficient and accurate glycan characterization via one-step dissociation. Fluorescence detection enables quantitation with high sensitivity independent of the glycan structure.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.

在化学系化学测量和成像项目的支持下，蒙特克莱尔州立大学的高金山和他的学生正在开展研究，寻求提高一类重要生物分子（称为聚糖）的同时表征和定量能力 - 碳水化合物修饰高博士的方法使用了质谱（MS），这是最广泛使用和最强大的化学分析工具之一。他正在开发用荧光化学“标签”标记聚糖分子的方法，以实现绝对定量并促进其结构表征。包括第一代大学生在内的本科生的参与预计将特别增强所提出的研究的影响。蒙特克莱尔州立大学是一所为西班牙裔服务的机构，主要是本科生机构，其研究内容已融入高博士的本科生和研究生课程中，使学生能够熟悉现代分析技术，并帮助他们为 STEM（科学、技术、工程和数学）领域的可能职业发展长期兴趣。高小组正在开发用于同步聚糖定量和表征的新型荧光自由基标签，以及新型多重同量异位荧光自由基标签用于同时对多个样品中的每个聚糖进行表征和定量。标签包含四个同样重要的化学特性：1) 荧光团有助于高灵敏度和可靠的定量；2)自由基前体预计会诱导系统、有效和可预测的断裂，以进行结构表征；3) 酰肼或羟胺基团选择性地靶向聚糖的还原末端；4) 吡啶或叔胺基团预计会显着增强MS 信号。这些定制标签旨在实现系统、高效和可预测的自由基介导的聚糖解离，从而通过一步解离实现高效、准确的聚糖表征。荧光检测能够以独立于聚糖结构的高灵敏度进行定量。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Gas-Phase Ion/Ion Reactions to Enable Radical-Directed Dissociation of Fatty Acid Ions: Application to Localization of Methyl Branching

气相离子/离子反应实现脂肪酸离子的自由基定向解离：在甲基支化定位中的应用

• DOI: 10.1021/acs.analchem.3c04510
• 发表时间: 2024-02
• 期刊: Analytical Chemistry
• 影响因子: 7.4
• 作者: Shenault, De’Shovon M.;Fabijanczuk, Kimberly C.;Murtada, Rayan;Finn, Shane;Gonzalez, L. Edwin;Gao, Jinshan;McLuckey, Scott A.
• 通讯作者: McLuckey, Scott A.

Development of mass spectrometric glycan characterization tags using acid‐base chemistry and/or free radical chemistry

使用酸碱化学和/或自由基化学开发质谱聚糖表征标签

• DOI: 10.1002/mas.21810
• 发表时间: 2022-09
• 期刊: Mass Spectrometry Reviews
• 影响因子: 6.6
• 作者: Murtada, Rayan;Finn, Shane;Gao, Jinshan
• 通讯作者: Gao, Jinshan

Mechanistic Study into Free Radical-Activated Glycan Dissociations through Isotope-Labeled Cellobioses

通过同位素标记的纤维二糖进行自由基激活聚糖解离的机理研究

• DOI: 10.1021/acs.analchem.2c04649
• 发表时间: 2023-02-07
• 期刊: Analytical Chemistry
• 影响因子: 7.4
• 作者: Fabijanczuk, Kimberly;Yu, Zaikuan Josh;Bakestani, Rose M.;Murtada, Rayan;Denton, Nicholas;Gaspar, Kaylee;Otegui, Tara;Acosta, Jose;Kenttamaa, Hilkka I.;Eshuis, Henk;Gao Jinshan
• 通讯作者: Gao Jinshan