Improved Design and Integration of SID Technology for Multiple MS Platforms

TR&D 1项目摘要。表面诱导解离（SID）已成为一种有效的工具，探测 蛋白质复合物在天然质谱（nMS）领域的拓扑结构。SID拥有独一无二的 有效切割非共价蛋白质复合物的界面以产生单体亚基的优点是， 以及低阶低聚物。此外，SID产品通常保持紧凑/高度结构化 构象，这样离子迁移率测量加上SID可以用来获得深入了解蛋白质 亚基结构通过碰撞截面（CCS）测量。本TR&D旨在改善SID 在nMS领域中使用的跨多个质量分析平台的技术（即四极杆时间- 飞行（QTOF）、OrbitrapTM和傅里叶变换离子回旋共振（FTICR）平台）。 目前的SID装置跨越具有至少十个DC电极的仪器的离子路径的3- 4cm。这些 电极电压可被调谐以将离子导向离轴表面（相对于在离子通道内的离子路径 用于激活离子-表面碰撞并随后将产物离子收集回 原始离子轨迹轴。透镜也可以被调整以引导离子通过装置而不激活， 允许进行标准MS实验（所谓的“飞越”模式）。 本TR&D的目的1是简化SID实验的调谐，同时保留或改善离子 在SID和“飞越”实验中的传输效率。一种新的SID设备， 碰撞表面和离子毯将被开发为比当前SID设备需要更少的电极，从而 简化了SID的调谐要求。离子地毯阵列的径向聚焦性质预计将有助于 离子传输效率。该设备将在更高分辨率的QTOF原型中实现。 目标2将采用目标1中取得的进展，以改善轨道阱平台内目前的SID性能 用于高分辨率nMS实验。对于电流，SID加速电压限值也约为70 V 轨道阱配置。与赛默飞世尔的合作将通过提供原理图来促进进展。 将独立电源集成到QExactive+上的电子板所需的信息 OSU的EMR平台和拟购买的QExactive HF UHMR平台。此次合作还将 促进将Aim 1开发的技术应用于Orbitrap Exactive平台内的离子光学。 目标3侧重于改进FTICR平台中的SID技术。目前，该平台拥有 m/z 6000的最大四极杆质量分离限度，对于选择较大的蛋白质来说太低 配合物目前在碰撞诱导解离的收集中存在降低的性能 由于原始冲突小区空间在SID设备和截断的冲突小区之间被分割而导致的碎片。 这两个缺点都将通过与供应商合作重新设计仪器来解决 （Bruker），如目的3中所述。

TR&D 1 Project Summary. Surface induced dissociation (SID) has emerged as an effective tool for probing the topology of protein complexes in the field of native mass spectrometry (nMS). SID has the unique advantage of efficiently cleaving the interfaces of noncovalent protein complexes to yield monomer subunits as well as small order oligomers. Additionally, SID products typically retain compact/highly structured conformations such that ion mobility measurements coupled with SID can be used to gain insights into protein subunit structure through collision cross section (CCS) measurements. This TR&D aims to improve SID technology across multiple mass analysis platforms that are used in the nMS field (namely quadrupole-time-of- flight (QTOF), OrbitrapTM, and Fourier transform ion cyclotron resonance (FTICR) platforms). Current SID devices span 3-4 cm of an instrument’s ion path with at least ten DC electrodes. These electrode voltages can be tuned to direct ions toward an off-axis surface (relative to the ion path within an instrument for the activating ion-surface collision and subsequent collection of product ions back onto the original ion trajectory axis. Lenses may also be tuned to guide ions through the device without activation, allowing for standard MS experiments to be conducted (so-called “flythrough” mode). Aim 1 of this TR&D is to simplify the tuning of SID experiments while retaining or improving ion transmission efficiencies in both SID and “fly-through” experiments. A new SID device containing an on-axis collision surface and ion carpet will be developed to require fewer electrodes than current SID devices, thereby simplifying tuning requirements for SID. The radial-focusing nature of the ion carpet array is expected to assist ion transmission efficiencies. The device will be implemented in a prototype higher resolution QTOF. Aim 2 will adopt advances made in Aim 1 to improve current SID performance within Orbitrap platforms for high resolution nMS experiments. There is also an SID acceleration voltage limit of roughly 70 V for current Orbitrap configurations. Collaboration with Thermo Fisher will facilitate progress by providing the schematic information necessary to incorporate independent power supplies into electronic boards on the QExactive+ EMR platform at OSU and QExactive HF UHMR platform proposed for purchase. This collaboration will also facilitate adapting technology developed from Aim 1 into the ion optics within Orbitrap Exactive platforms. Aim 3 focuses on improving SID technology in an FTICR platform. Currently, this platform has a maximum quadrupole mass isolation limit of m/z 6000, which is too low for selection of larger protein complexes. There is currently a diminished performance in the collection of collision induced dissociation fragments due to the original collision cell space being split between an SID device and truncated collision cell. Both of these shortcomings will be addressed through instrument redesign in collaboration with the vendor (Bruker) as described in Aim 3.