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

在化学系化学测量和成像项目的支持下，高金山和他在蒙特克莱尔州立大学的学生们正在进行研究，寻求提高同时表征和定量一类重要的生物分子（称为聚糖）的能力。聚糖是修饰生物系统中许多蛋白质的碳水化合物修饰，在生物医学科学中具有重要意义。高博士的方法使用质谱（MS），这是化学分析中使用最广泛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。拟议的研究的影响预计将通过本科生的参与，包括第一代大学生和科学代表性不足的群体的成员-蒙特克莱尔州立大学是一个西班牙裔服务机构和私立本科院校的增强。 高博士的本科和研究生课程中融入了研究元素，使学生能够熟悉现代分析技术，并帮助他们培养对STEM职业的长期兴趣（科学，技术，工程和数学）领域。高小组正在开发新型荧光自由基标签，用于同时聚糖定量和表征，以及新型多路复用，同量异序荧光自由基标签，用于同时表征和定量来自多个样品的每种聚糖。这些标签包含四个同等重要的化学特征：1）荧光团促进高度灵敏和可靠的定量; 2）自由基前体预期诱导系统的、有效的和可预测的片段化以用于结构表征; 3）酰肼或羟胺基团选择性地靶向聚糖的还原末端;和4）吡啶或叔胺基团预期显著增强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和准确的聚糖表征。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知识价值和更广泛的影响审查标准进行评估而被认为值得支持。

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.