CAREER: Three Region nLC-MS/MS Analysis: A Novel and Rapid Approach to Evaluate Molecular Penetration

在这个CAREER项目中，圣母大学的Amanda Hummon教授和她的学生将开发一种新的质谱分析方法，用于分析细胞培养物中的化合物。该方法将以比目前任何其他方法更快和更高通量的方式提供更多的化学和空间信息。作为这个职业教育项目的一部分，Hummon教授和她的学生们将制定一个项目，专注于在STEM领域招募和留住女性，每年组织三个当地女童子军的实验室参观，每学期参加两个论坛，讨论对女性产生不成比例影响的问题。这个项目将鼓励年轻的女学生参与科学事业。本研究将通过质谱法研究化合物对三维细胞培养模型的渗透。采用串联胰蛋白酶化和纳米液相色谱-电喷雾串联质谱（nLC-MS/MS）相结合的方法，可以绘制化合物及其代谢物的空间保真度分布图。虽然以前的研究利用计算机模拟来预测分子穿透细胞，但测量分子仍然是评估新物种有效性的唯一可靠方法。该方法将用化合物伊立替康、奥沙利铂和细胞凋亡成像探针进行测试。该奖项由化学部门的化学测量和成像项目与生物基础设施部门的生物研究仪器开发项目联合颁发。

In this CAREER project, Professor Amanda Hummon and her students at the University of Notre Dame will develop a new mass spectrometric methodology for the analysis of chemical compounds in cell cultures. The approach will provide much greater chemical and spatial information in a faster and higher-throughput fashion than is currently available with any other methodology. As part of this CAREER educational program, Professor Hummon and her students will develop a program focused on the recruitment and retention of women in STEM fields by organizing lab tours with three local Girl Scout troops per year and participation in two forums per semester to discuss issues that disproportionally affect women. This program will encourage the participation of younger female students in science careers.This research will study the penetration of chemical compounds into three dimensional cell culture models via mass spectrometry. Using a combination of serial trypsinization and nano Liquid Chromatography-Electrospray Tandem Mass Spectrometry (nLC-MS/MS), the distribution of a compound and its metabolites can be mapped with spatial fidelity. While previous studies have utilized computer simulations to predict molecular penetration through cells, measurement of the molecules remains the only reliable way to evaluate the effectiveness of a new species. The approach will be tested with the compounds irinotecan, oxaliplatin, and apoptotic imaging probes. This award from the Chemical Measurements and Imaging program in the Division of Chemistry is made jointly with the Instrument Development for Biological Research program in the Division of Biological Infrastructure.