Ultrasensative analysis of biological molecules

生物分子的超灵敏分析

• 批准号: RTI-2021-00055
• 负责人: Macauley, Matthew
• 金额: $ 7.74万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718777
• 关键词: Ultrasensative; analysis; biological; molecules

Biomolecules such as cytokines, chemokines, and phosphorylated cellular signaling components can elicit cellular responses at very low concentrations due to their ability to propagate signals. Such low concentrations can make detection of these critical mediators challenging, particularly from complex biological specimens. Low levels of biomolecules, limited amounts of a biological specimen, and the need to do multiple timepoints are inherent challenges in measuring the levels of bioanalytes from biological samples. Ultrasensitive detection of bioanalytes using electrochemiluminescence overcomes low concentrations because the limits of detection for this method far exceed (100X) traditional approaches that have a chromogenic readout (e.g. ELISA). Combining ultrasensitive detection with multiplexing provides a solution by enabling many bioanalytes to be measured in a single small aliquot of a precious sample. Our team of investigators at the University of Alberta have a great need for sensitive measurements. The Macauley lab studies immunomodulatory receptors that bind carbohydrates, with particular interest in immune cells in the brain, called microglia; the Richard lab studies the impact of nutrition on immune homeostasis; the Willing lab studies the role microbes play in intestinal health in livestock species; and the Field lab studies the roles that fats in the postnatal diet play in supporting immune cell development. Making fundamental discoveries in these areas requires precise and sensitive detection of biomolecules, such as: cytokines, chemokines, phosphorylated signaling molecules, antibodies, and metabolites. The only openly available electrochemiluminescence reader on the University of Alberta campus recently reached the end of its usable life and is no longer operational because parts to fix this instrument are not available. This has left many research groups stranded with the inability to carry out these measurements. Accordingly, there is an urgent need for a reliable ultrasensitive electrochemiluminescence reader housed in a shared space. Having these capabilities is critical for the research programs of our team as well as dozens of other research groups at the University of Alberta. The previous instrument, housed in the Alberta Diabetes Institute molecular core facility, was well used by many groups across campus with an average of over 200 plates run per year. Therefore, we are requesting funds for an ultrasensitive electrochemiluminescence instrument for detecting bioanalytes from complex biological samples to accelerate the discovery of mechanisms controlling immune cell function, factors impacting host-microbial interactions, and the mechanisms by which nutrition regulates immune homeostasis and development. In addition to generating experimental data that are critical for our research programs, acquiring an ultrasensitive electrochemiluminescence reader will provide valuable learning opportunities for our trainees.

生物分子如细胞因子、趋化因子和磷酸化的细胞信号传导组分由于其传播信号的能力而可以在非常低的浓度下引发细胞应答。如此低的浓度可能使这些关键介质的检测具有挑战性，特别是从复杂的生物样本中。低水平的生物分子、有限量的生物样本以及需要在多个时间点进行测量是测量来自生物样品的生物分析物水平的固有挑战。使用电化学发光的生物分析物的超灵敏检测克服了低浓度，因为该方法的检测极限远远超过（100倍）具有显色读出的传统方法（例如ELISA）。将超灵敏检测与多路复用相结合提供了一种解决方案，使许多生物分析物能够在珍贵样品的单个小等分试样中进行测量。我们在阿尔伯塔大学的研究团队非常需要灵敏的测量。Macauley实验室研究结合碳水化合物的免疫调节受体，特别关注大脑中的免疫细胞，称为小胶质细胞; Richard实验室研究营养对免疫稳态的影响; Willing实验室研究微生物在家畜肠道健康中的作用; Field实验室研究产后饮食中的脂肪在支持免疫细胞发育中的作用。在这些领域进行基础性发现需要精确和灵敏地检测生物分子，例如：细胞因子，趋化因子，磷酸化信号分子，抗体和代谢物。阿尔伯塔大学校园内唯一一台公开使用的电化学发光阅读器最近达到了使用寿命，由于没有固定该仪器的部件，因此不再运行。这使得许多研究小组无法进行这些测量。因此，迫切需要一种容纳在共享空间中的可靠的超灵敏电化学发光读取器。拥有这些能力对于我们团队的研究计划以及阿尔伯塔大学的其他数十个研究小组至关重要。以前的仪器，设在阿尔伯塔糖尿病研究所分子核心设施，是很好地使用了许多团体在校园内，平均每年超过200板运行。因此，我们正在申请资金，用于从复杂的生物样品中检测生物分析物的超灵敏电化学发光仪器，以加速发现控制免疫细胞功能的机制，影响宿主-微生物相互作用的因素，以及营养调节免疫稳态和发育的机制。除了生成对我们的研究计划至关重要的实验数据外，获得超灵敏的电化学发光阅读器将为我们的学员提供宝贵的学习机会。