Increasing the Selectivity of Hybrid Mass Spectrometry using Multidimensional Ion Mobility Spectrometry

总结 目前用于快速表征生物分子的结构、异质性、质量和相似性的工具 不能提供许多应用所需的选择性水平，包括生物芯片开发， 生产和监管。质谱法（MS）和离子迁移率（IM）对于 由于其速度、灵敏度、对样品异质性的耐受性， 可伸缩性这个项目将产生一个新的平台，它将实现更高的性能和更高的维度 生物分子的表征，包括完整的蛋白质和多蛋白复合物。新测量结果 使用现有的IM-MS平台是不可能的。该平台将使用模块化 分离、选择和捕获类天然离子的组件，这些组件将组合成一个大型模块阵列 (Aim 1）并与离子结构的互补探针整合（目的2）。该平台将使多个 探测离子的形状、稳定性、内部相互作用和反应性的分析维度。的 这些分析维度之间的正交性将增加测量的选择性， 这些目标将使用模型（目标3）进行评估 蛋白质，蛋白质-配体复合物，和蛋白质-蛋白质复合物，以评估这一潜力， 平台，以回答生物医学研究中的挑战性问题。更一般地说，这项研究将改善 更高维IM-MS实验的可行性，并将提高正交性的理解 比在分析的每个维度之间可以实现的要多。该项目受益于模块化 组件和快速原型，但与这项研究的结果，我们预计，这种灵活性， 实验设计和这些测量的选择性可以被转换为 更加用户友好（尽管模块化程度较低），从而对生物医学科学产生广泛影响， 生物制药行业。

SUMMARY Current tools for rapidly characterizing the structures, heterogeneity, quality, and similarity of biomolecules do not provide the level of selectivity needed for many applications, including biotherapeutic development, production, and regulation. Mass spectrometry (MS) and ion mobility (IM) are particularly appealing for addressing these unmet needs due to their speed, sensitivity, tolerance for sample heterogeneity, and scalability. This project will result in a new platform that will enable higher-performance and higher-dimensional characterization of biomolecules, including intact proteins and multiprotein complexes. The new measurements enabled by this platform are not possible using existing IM-MS platforms. This platform will use modular components to separate, select, and trap native-like ions, which will be combined into a large array of modules (Aim 1) and integrated with complementary probes of ion structure (Aim 2). This platform will enable multiple dimensions of analysis that probe the shapes, stabilities, internal interactions, and reactivity of the ions. The orthogonality between these dimensions of analysis will increase the selectivity of measurements, without compromising the underlying strengths of IM and MS. These aims will be (Aim 3) evaluated using model proteins, protein-ligand complexes, and protein-protein complexes in order to assess the potential of this platform to answer challenging problems in biomedical research. More generally, this research will improve the feasibility of higher-dimensional IM-MS experiments and will improve the understanding of the orthogonality than can be achieved between each dimensions of analysis. This project benefits greatly from modular components and rapid prototyping, but with the outcomes of this research, we anticipate that this flexibility of experimental design and the selectivity of these measurements can be translated to architectures that are more user friendly (albeit less modular) and thereby have broad impact across the biomedical sciences and biopharmaceutical industries.