High performance mass spectrometry: applications for the Cambridge biological sciences community

高性能质谱：剑桥生物科学界的应用

• 批准号: BB/W019620/1
• 负责人: Kathryn Lilley
• 金额: $ 37.64万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019620%2F1
• 关键词: performance; mass; spectrometry; applications; Cambridge

The Cambridge Centre for Proteomics (CCP) is a well-established mass spectrometry core facility in the Department of Biochemistry, University of Cambridge. CCP primarily offers proteomics services. Proteomics is the study of all proteins in a sample which could be proteins extracted from plant leaves, human parasites, bacteria, fungi and clinical samples such as plasma and urine. Researchers generally wish to know what proteins are present in their samples, but importantly the abundance of the proteins and how this changes in response to stress, drug treatment or disease state. Also, researchers may want information on the post translational modifications in the proteome and how this changes upon perturbation, with phosphorylation being the most popular modification studied.CCP uses mass spectrometry as its major tool to interrogate the proteome. This instrument measures the mass of ions introduced typically via a liquid chromatography device, together referred to as LC-MS. From these mass measurements, both the identity of proteins present and their amount can be calculated. Mass spectrometry can also be used to characterise other biomolecules such as sugars, lipids and nucleic acids. CCP is a very popular and busy facility. It currently has access to two mass spectrometers. Its Thermo QExactive is 9 years old and is very unreliable. It is covered by a contract to fix it if it breaks, but its manufacturer will discontinue the contract for this instrument in the next 2 years. This instrument is insensitive compared with more modern instruments. CCP also has access to 55% of the time of a 5-year-old Thermo Fusion Lumos Tribrid, which is more sensitive and a very good instrument to carry out a form of quantitative proteomics known as isobaric tagging. This instrument lacks sensitivity compared with newer instruments and is able to separate ions introduced into the instrument based on their mobility in the gas phase. Together, these instruments do not provide CCP with the capacity to process samples submitted by its users.The need for a more up to data mass spectrometer has become critical. CCP supports its staff and equipment through fee for service and grant funding. CCP simply cannot sustain its services and will be faced with significant cuts. Moreover, it has increasing demands from it users including those with limited sample amounts or who need more elaborate characterisation of post translational modification.Other facilities in Cambridge are reserved for the institutes in which they sit and have no spare capacity to assist CCP.The requested LC-MS, Bruker TimsTOF Pro 2 is much more sensitive than CCP's current instruments. It processes samples more quickly and hence will increase the capacity of CCP and reduce sample turnround significantly. It also is equipped with an ion mobility functionality which assists in both sensitivity and distinguishing very similar molecules based on their cross sectional area as gaseous ions. This instrument excels at quantitative label free proteomics approaches that do not require the use of isobaric tagging, and is highly complementary to what is currently available here. Its ion mobility function also helps distinguish other classes of biomolecules with similar masses but subtly different behaviours within ion mobility. There is no similar LC-MS instrument in CCP or indeed in Cambridge.This LC-MS system will support all CCP users from across the University of Cambridge, outside research institutes, users form other academic institutions and users from industry. The proposed equipment, will not only allow them to carry out their planned work, but enable new avenues of research which have been hitherto impossible with existing instrumentation. The support from the University of Cambridge means that this proposal requests less than half of the cost of the TimsTOF Pro 2, and thus represents value for money.

剑桥蛋白质组学中心（CCP）是剑桥大学生物化学系一个完善的质谱核心设施。CCP主要提供蛋白质组学服务。蛋白质组学是对样品中所有蛋白质的研究，样品可以是从植物叶片、人类寄生虫、细菌、真菌和临床样品（如血浆和尿液）中提取的蛋白质。研究人员通常希望知道他们的样品中存在什么蛋白质，但更重要的是蛋白质的丰度，以及这种蛋白质在压力、药物治疗或疾病状态下是如何变化的。此外，研究人员可能想要了解蛋白质组的翻译后修饰以及这种修饰在扰动下如何变化，磷酸化是研究中最流行的修饰。CCP使用质谱法作为其主要工具来询问蛋白质组。该仪器测量通常通过液相色谱装置引入的离子的质量，统称为LC-MS。从这些质量测量中，可以计算出存在的蛋白质的身份及其数量。质谱法也可用于表征其他生物分子，如糖、脂质和核酸。CCP是一个非常受欢迎和繁忙的设施。它目前有两个质谱仪。它的Thermo QExactive已经用了9年了，非常不可靠。如果它坏了，它是有修理合同的，但它的制造商将在未来两年内终止该仪器的合同。与更现代的仪器相比，这台仪器不灵敏。CCP还可以使用已有5年历史的Thermo Fusion Lumos Tribrid的55%的时间，这是一种更敏感的工具，也是一种非常好的工具，可以进行一种被称为等压标记的定量蛋白质组学。与较新的仪器相比，该仪器缺乏灵敏度，并且能够根据其在气相中的迁移率分离引入仪器的离子。总之，这些仪器不提供CCP处理其用户提交的样品的能力。对更先进的质谱仪的需求已经变得至关重要。CCP通过服务费和赠款资助来支持其员工和设备。CCP根本无法维持其服务，将面临大幅削减。此外，它的用户需求越来越大，包括那些样本量有限或需要更详细地描述翻译后修饰的用户。剑桥的其他设施是为他们所在的研究所保留的，没有多余的能力来帮助CCP。所要求的LC-MS，布鲁克TimsTOF Pro 2比CCP目前的仪器更敏感。它处理样品的速度更快，因此将增加CCP的能力，并显著减少样品周转率。它还配备了离子迁移功能，有助于灵敏度和区分非常相似的分子，基于它们作为气体离子的横截面积。该仪器擅长于不需要使用等压标记的定量无标签蛋白质组学方法，并且与目前可用的方法高度互补。它的离子迁移功能也有助于区分其他种类的生物分子具有相似的质量，但在离子迁移中有细微的不同行为。在CCP或剑桥没有类似的LC-MS仪器。该LC-MS系统将支持剑桥大学、外部研究机构、其他学术机构和行业用户的所有CCP用户。拟议中的设备不仅可以让他们进行计划中的工作，还可以为现有仪器迄今无法实现的研究开辟新的途径。剑桥大学的支持意味着该提案所需的成本不到TimsTOF Pro 2的一半，因此代表了物有所值。