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

在蒙特克莱尔州立大学化学系化学测量与成像项目的支持下，高金山和他的学生们正在进行研究，寻求提高同时表征和定量重要生物分子聚糖的能力。聚糖是一种修饰生物系统中许多蛋白质的碳水化合物，在生物医学科学中具有重要意义。高博士的方法使用了质谱（MS），这是化学分析中最广泛使用和最强大的工具之一。具体来说，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。预计通过本科生的参与，包括第一代大学生和科学界代表性不足的群体的成员，拟议的研究的影响将得到加强-蒙特克莱尔州立大学是一所为西班牙裔服务的机构和主要的本科机构。该研究的元素被整合到高博士的本科和研究生课程中，使学生熟悉现代分析技术，并帮助他们在STEM（科学、技术、工程和数学）领域发展长期的职业兴趣。Gao团队正在开发新型荧光自由基标签，用于同时定量和表征多糖，以及新型多路、等压荧光自由基标签，用于同时表征和定量来自多个样品的每种聚糖。该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)自由基前体有望诱导系统、高效和可预测的碎片化，用于结构表征；3)一个肼基或氧胺基选择性地靶向聚糖的还原端；4)吡啶或叔胺基团有望显著增强质谱信号。这些量身定制的标签旨在实现系统，高效和可预测的自由基介导的聚糖解离，通过一步解离实现高效和准确的聚糖表征。荧光检测使定量具有高灵敏度，独立于聚糖结构。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.