Infrastructure to Support Ultra-High Separation of Biological Samples

支持生物样品超高分离度的基础设施

• 批准号: RTI-2022-00531
• 负责人: GolemiKotra, Dasantila
• 金额: $ 10.93万
• 依托单位: York University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=742964
• 关键词: Infrastructure; Support; Ultra; Separation; Biological

In this proposal, we aim to establish an infrastructure that is capable of ultra-high separation of complex biological samples in a highly reproducible and quantitative manner. Currently, this capacity is lacking in my laboratory. Therefore, we are requesting an Ultra Performance Liquid Chromatography system (UPLC). The proposed instrument is equipped with a fully integrated "on-line" mass spectrometer (UPLC-MS) orthogonal to an optical detector. The research in my laboratory focuses on understanding the bacterial response to cell wall damage. The underlying theme of the research program to be supported by the acquisition of the state-of-the-art UPLC-MS is the separation and characterization of complex bacterial biomolecules. The proposed infrastructure will impact the research on four fronts: First, it will enable the isolation and analysis of complex biological samples, such as cell wall components of bacteria, full identification of modified proteins from cell extracts, and separation of enzymatic products. Second, it will minimize impurities and thus facilitating the downstream processing of the samples. Third, it will increase the confidence in data acquisition and quantification. Lastly, the integrated mass detector will significantly decrease the reliance on specialized analytical lab services to determine the identity of our analytes, thus optimizing the research time and decreasing its cost. Therefore, UPLC-MS is indispensable to the advancement of the research in my group. In addition, the advantages provided by UPLC-MS, such as highly reproducible flowrates, high-resolution separations and highly reliable quantitative data, will meet today's research standards required from granting agencies and journals for an unambiguous reproducibility and quality of analytical separations and quantitative analyses of the experimental data. Moreover, this instrument will provide an indispensable training platform to the HQPs in analytical skills, which are in high demand in research, pharmaceutical and diagnostic settings. As such, the trainees will be marketable to the current needs of industry and research settings. Currently, the training of HQPs is stifled. Since its inception in 2004 (the year I joined YorkU), my research program revolves around biological molecules that need to be separated, isolated, identified and characterized. So, the requested instrument is crucial in supporting the essence of my research, its expansion and impact on the understanding of bacterial response to cell wall damage. Above all, throughout these years at YorkU, my laboratory has trained a diverse body of trainees; coming from different backgrounds and walks of life. Many had dreams of becoming professors or doctors. Others were mothers that wanted a better life for their children and be a role model to them. It is fulfilling to see that the training in my laboratory has enabled them to fulfill their aspirations.

在这项提案中，我们的目标是建立一个基础设施，能够以高度可重复和定量的方式超高分离复杂的生物样品。目前，我们的实验室缺乏这种能力。因此，我们需要一个超高效液相色谱系统（UPLC）。建议的仪器配备了一个完全集成的“在线”质谱仪（UPLC-MS）正交的光学检测器。 我实验室的研究重点是了解细菌对细胞壁损伤的反应。通过收购最先进的UPLC-MS支持的研究计划的基本主题是复杂细菌生物分子的分离和表征。拟议的基础设施将在四个方面影响研究：首先，它将能够分离和分析复杂的生物样品，如细菌的细胞壁成分，从细胞提取物中完全鉴定修饰的蛋白质，以及分离酶产物。其次，它将最大限度地减少杂质，从而促进样品的下游处理。第三，它将增加数据采集和量化的信心。最后，集成的质量检测器将大大减少对专业分析实验室服务的依赖，以确定我们分析物的身份，从而优化研究时间并降低成本。因此，超高效液相色谱-质谱联用技术对本课题组的研究进展是必不可少的。此外，UPLC-MS提供的优势，如高重现性的流速，高分辨率的分离和高度可靠的定量数据，将满足当今的研究标准，要求授权机构和期刊明确的重现性和分析分离的质量和实验数据的定量分析。 此外，这一工具将为总部质量保证人员提供分析技能方面的不可或缺的培训平台，而这在研究、制药和诊断环境中需求量很大。因此，受训人员将适应工业和研究环境的当前需求。目前，对HQP的培训受到抑制。 自2004年（我加入约克大学的那一年）成立以来，我的研究项目围绕着需要分离，分离，鉴定和表征的生物分子。因此，所要求的仪器对于支持我的研究的本质、其扩展以及对理解细菌对细胞壁损伤的反应的影响至关重要。最重要的是，在约克大学的这些年里，我的实验室培养了一批来自不同背景和各行各业的学员。许多人梦想成为教授或医生。其他人是母亲，希望为自己的孩子过上更好的生活，并成为他们的榜样。看到在我的实验室里的培训使他们能够实现自己的愿望，我感到很满足。