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

在这个职业项目中，圣母大学的阿曼达·胡蒙教授和她的学生将开发一种新的质谱学方法来分析细胞培养中的化学成分。与目前任何其他方法相比，该方法将以更快和更高吞吐量的方式提供更多的化学和空间信息。作为这一职业教育计划的一部分，Hummon教授和她的学生将制定一个计划，重点是招募和留住STEM领域的女性，方法是每年与三名当地女童子军组织实验室参观，并每学期参加两个论坛，讨论对女性影响不成比例的问题。该计划将鼓励更年轻的女学生参与科学事业。本研究将通过质谱学研究化合物在三维细胞培养模型中的渗透。利用连续胰酶消化和纳米液相色谱-电喷雾串联质谱仪(NLC-MS/MS)相结合的方法，可以以空间保真度绘制化合物及其代谢物的分布图。虽然之前的研究利用计算机模拟来预测分子对细胞的渗透，但对分子的测量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。该奖项由化学部的化学测量和成像计划与生物基础设施部的生物研究仪器开发计划联合颁发。

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.