Infrastructure to Support Ultra-High Separation of Biological Samples

在这项提议中，我们的目标是建立一个能够以高度可重复性和定量的方式对复杂生物样本进行超高分离的基础设施。目前，我的实验室缺乏这种能力。因此，我们需要一套超高性能的高效液相色谱系统(UPLC)。该仪器配备了一个与光学检测器垂直的全集成“在线”质谱仪(UPLC-MS)。我实验室的研究重点是了解细菌对细胞壁破坏的反应。收购最先进的UPLC-MS将支持这项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：第一，它将能够分离和分析复杂的生物样本，如细菌的细胞壁成分，从细胞提取物中全面鉴定修饰蛋白质，以及分离酶产品。其次，它将最大限度地减少杂质，从而方便样品的下游处理。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显著减少对专门的分析实验室服务的依赖，以确定我们的分析物的身份，从而优化研究时间并降低其成本。因此，UPLC-MS对于推进本课题组的研究是不可或缺的。此外，UPLC-MS提供的优势，如高重现性、高分辨率分离和高度可靠的定量数据，将满足授权机构和期刊对实验数据的分析分离和定量分析的明确重复性和质量的当今研究标准。此外，该仪器将为HQP提供一个不可或缺的分析技能培训平台，这些技能在研究、制药和诊断环境中的需求量很大。因此，受训人员将适应当前工业和研究环境的需求。目前，HQP的培训被扼杀了。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展开。因此，所要求的仪器对于支持我的研究的本质、其扩展和对了解细菌对细胞壁破坏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，她们希望孩子过上更好的生活，并成为他们的榜样。看到我的实验室的培训使他们能够实现他们的抱负，我感到很满意。

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.