Improved Design and Integration of SID Technology for Multiple MS Platforms

研发1个项目总结。表面诱导解离(SID)是一种有效的探测工具 蛋白质复合体的拓扑学在自然质谱学领域中的应用。SID拥有唯一的 有效地切割非共价蛋白质复合体的界面以产生单体亚基的优势 以及小顺序齐聚物。此外，SID产品通常保持紧凑/高度结构化 离子迁移率测量与SID相结合的构象可以用来洞察蛋白质 通过碰撞截面(CCS)测量亚基结构。这项研发旨在改善SID 跨NMS领域使用的多个质量分析平台的技术(即四极时间 飞行(QTOF)、OrbitRapTM和傅里叶变换离子回旋共振(FTICR)平台)。 目前的SID设备具有至少10个直流电极，跨越仪器离子路径的3-4厘米。这些 可以调整电极电压以将离子引向离轴表面(相对于 用于激活离子-表面碰撞并随后将产品离子收集回 原始离子轨迹轴。透镜也可以被调谐以在不激活的情况下引导离子通过设备， 允许进行标准的MS实验(所谓的“飞行”模式)。 本研发的目标1是简化SID实验的调整，同时保留或改进离子 在SID和“穿越”实验中的传输效率。一种包含轴上驱动器的新型SID设备 碰撞表面和离子地毯将被开发成比目前的SID器件需要更少的电极，从而 简化SID的调整要求。离子地毯阵列的径向聚焦特性有望有助于 离子传输效率。该装置将在更高分辨率的QTOF样机上实现。 Aim 2将采用Aim 1中的改进，以改进Orbitrap平台中当前的SID性能 用于高分辨率NMS实验。此外，电流的SID加速电压限制约为70V 轨道诱骗配置。与Thermo Fisher的合作将通过提供原理图来促进进展 将独立电源整合到QExactive+上的电子板中所需的信息 OSU的EMR平台和QExactive高频UHMR平台建议购买。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，该平台有一个 最大四极杆质量分离极限为m/z 6000，对于选择更大的蛋白质来说太低了 复合体。目前，碰撞诱导解离的收集性能有所下降 由于原始冲突信元空间在SID设备和被截断的冲突信元之间被分割而产生的碎片。 这两个缺点将通过与供应商合作重新设计仪器来解决 (布鲁克)，如目标3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.