Infrastructure to Support Ultra-High Separation of Biological Samples

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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.