Development of High-throughput Multidimensional Collision Induced Unfolding Technology

在化学系化学测量和成像项目的支持下，密歇根大学的布兰登·鲁托洛教授和他的团队正在开发强大的新工具，用于表征蛋白质和RNA的三维结构-这两种生物大分子在生命过程中的关键类别。具体来说，Ruotolo小组正在扩展“碰撞诱导解折叠”（CIU）的能力，这是一种通过快速去除正常环境中生物分子周围的水来研究蛋白质结构的方法，然后诱导它们的解折叠，同时使用称为离子迁移率质谱（IM-MS）的最先进工具监测它们的大小和形状。这种方法有可能彻底改变生物分子稳定性评估的细节和采集速度。潜在的应用包括下一代工程生物治疗药物的快速分析。这项工作将为对化学测量和制药科学之间的接口研究感兴趣的学生提供出色的培训。根据该奖项，密歇根大学的Ruotolo研究团队旨在解决三个主要研究目标。 首先，该团队将致力于建立用于操作循环型IM-MS仪器的改进方法，以改善天然生物分子结构的保留，碰撞截面（CCS）校准精度和高阶IM实验（例如，IM-CIU和CUN）。其次，与此同时，密歇根团队将奋进开发下一代软件，该软件包含自动峰值拾取、特征跳过、机载CCS校准和对CIUN数据类型的支持等功能。Ruotolo教授和他的学生们的目标是利用这些改进的IM-MS和CIU工具来研究抗体和大RNA，努力建立支持对生物技术领域具有重要意义的尖端生物分子工程活动的测定方法。该奖项反映了NSF的法定使命，并通过使用基金会的智力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, Professor Brandon Ruotolo and his group at the University of Michigan are developing powerful new tools for characterizing the three-dimensional structures of proteins and RNA – both key classes of biomacromolecules in life processes. Specifically, the Ruotolo group is expanding the capabilities of "collision induced unfolding" (CIU), an approach that studies protein structures by quickly removing the water which surrounds biological molecules in their normal environment, then induces their unfolding while monitoring their size and shape using a state-of-the-art tool known as ion mobility-mass spectrometry (IM-MS). The approach has the potential to revolutionize the detail and acquisition speed with which biomolecular stability can be evaluated. Potential applications include the rapid analysis of next-generation engineered biotherapeutics. The work will provide excellent training for students interested in research at the interface between chemical measurements and the pharmaceutical sciences.Under this award, the Ruotolo research team at the University of Michigan aims to addresses three main research goals. Firstly, the team will work on building improved methods for the operation of cyclic-type IM-MS instruments for improved retention of native-like biomolecular structure, collision cross-section (CCS) calibration accuracy, and higher-order IM experiments (e.g., IM-CIU and CIUN). Secondly, and in parallel, the Michigan team will endeavor to develop next-generation software that incorporates features for automated peak picking, feature skipping, on-board CCS calibration, and support for CIUN data types. Then Professor Ruotolo and his students aim to leverage these improved IM-MS and CIU tools to study antibodies and large RNAs in an effort to build assays that support cutting-edge biomolecular engineering campaigns of significance to the biotechnology sector.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.