A cellenONE BSC to enhance UK capability for single-cell proteomics

cellenONE BSC 可增强英国单细胞蛋白质组学能力

• 批准号: BB/W019744/1
• 负责人: Edward Emmott
• 金额: $ 23.23万
• 依托单位: University of Liverpool
• 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%2FW019744%2F1
• 关键词: cellenONE; BSC; enhance; UK; capability

Single-cell studies are transforming many areas of biological research; these include studies of infection, differentiation, construction of cell atlases, and more. Single-cell proteomics is a rapidly emerging addition to the single-cell toolset, with single-cell proteomics by mass spectrometry capable of measuring the abundance of thousands of proteins across hundreds to thousands of single-cells. By acquiring state-of-the-art instrumentation for single-cell proteomics sample preparation, this proposal seeks to enhance UK capabilities in single-cell proteomics. Specific applications of this instrument include:i) Further development and application of single-cell proteomics. The cellenONE increases the sensitivity of single-cell proteomics by minimizing sample losses during processing. We will exploit this increased sensitivity to target specific proteins of interest for reproducible analysis across single-cells, study post-translational modifications at single-cell level, as well as apply this system to develop new methods for spatial and subcellular single-cell 'omics.ii) Cell/picolitre dispensing for other single-cell approaches. In this proposal we specifically apply this instrument to single-cell western blotting, providing information on protein molecular weight, and more sensitive comparative analysis as the direct introduction of cells into nanowells permits multiple samples to be analysed per chip, minimizing batch effects. Similarly, the cellenONE instrument will be applied to single-cell RNAseq studies, where the gentle acoustic dispensing used by the system can overcome cell-size-based biases or problems analysing fragile cells in existing approaches.We propose to bring this novel instrumentation which enables automation of the entire single-cell proteomics workflow to the Centre for Proteome Research (CPR) at the University of Liverpool (UoL). Making full use of our existing suite of LC-MS/MS instruments, and using our established cost recovery model which ensures long-term maintenance and support (supported by LIV-SRF which oversees research facilities at UoL), this instrumentation will be made available to researchers working across BBSRC priority areas, both locally and nationally. LIV-SRF provides financial support to maintain and develop open facilities such as the CPR, including long-term support of staff. LIV-SRF also awards 'LIV-SRF Vouchers' which provide academics, especially ECRs with matched funds to support facility usage and develop preliminary data, to support future funding or fellowship applications. All five current cellenONE systems in academic labs in the UK are located in London or the East of England, and none are currently used for single-cell proteomics. This instrument will be applied to multiple BBSRC-funded research areas, and our demonstration projects and letters of support for this system have focused on research impacting infection biology, fundamental cell signaling research, and immunity. This instrument will enable us to both directly contribute to these research areas, as well as support training for other UK academics and industry users to establish single-cell proteomics at their own institutions.

单细胞研究正在改变生物学研究的许多领域；这些包括感染、分化、细胞图谱构建等方面的研究。单细胞蛋白质组学是单细胞工具集的一个快速新兴的补充，通过质谱分析的单细胞蛋白质组学能够测量数百到数千个单细胞中数千种蛋白质的丰度。通过购买最先进的单细胞蛋白质组学样品制备仪器，该提案旨在增强英国在单细胞蛋白质组学方面的能力。该仪器的具体应用包括：i)单细胞蛋白质组学的进一步开发和应用。 cellenONE 通过最大限度地减少处理过程中的样品损失来提高单细胞蛋白质组学的灵敏度。我们将利用这种对目标特定蛋白质的敏感性增加，进行单细胞的可重复分析，研究单细胞水平的翻译后修饰，并应用该系统开发空间和亚细胞单细胞组学的新方法.ii) 其他单细胞方法的细胞/皮升分配。在本提案中，我们特别将该仪器应用于单细胞蛋白质印迹，提供蛋白质分子量信息和更灵敏的比较分析，因为将细胞直接引入纳米孔允许每个芯片分析多个样品，从而最大限度地减少批次效应。同样，cellenONE 仪器将应用于单细胞 RNAseq 研究，其中系统使用的温和声学分配可以克服基于细胞大小的偏差或现有方法中分析脆弱细胞的问题。我们建议引入这种新颖的仪器，使利物浦大学 (UoL) 蛋白质组研究中心 (CPR) 的整个单细胞蛋白质组工作流程的自动化。充分利用我们现有的 LC-MS/MS 仪器套件，并使用我们既定的成本回收模型来确保长期维护和支持（由负责监督伦敦大学研究设施的 LIV-SRF 提供支持），该仪器将可供使用面向在本地和国家 BBSRC 优先领域工作的研究人员。 LIV-SRF 提供财务支持来维护和开发 CPR 等开放设施，包括对员工的长期支持。 LIV-SRF 还颁发“LIV-SRF 优惠券”，为学术界（尤其是 ECR）提供配套资金，以支持设施使用和开发初步数据，以支持未来的资助或奖学金申请。英国学术实验室目前所有五个 cellenONE 系统均位于伦敦或英格兰东部，目前没有一个系统用于单细胞蛋白质组学。该仪器将应用于多个 BBSRC 资助的研究领域，我们对该系统的示范项目和支持信重点关注影响感染生物学、基础细胞信号研究和免疫的研究。该仪器将使我们能够直接为这些研究领域做出贡献，并支持其他英国学者和行业用户在自己的机构建立单细胞蛋白质组学的培训。