Ultrasensative analysis of biological molecules

生物分子，如细胞因子、趋化因子和磷酸化的细胞信号成分，由于其传播信号的能力，可以在非常低的浓度下引发细胞反应。如此低的浓度会使这些关键介体的检测具有挑战性，特别是从复杂的生物标本中。生物分子水平低、生物样本数量有限以及需要进行多个时间点测试是测量生物样本中生物分析物水平的内在挑战。使用电化学发光法对生物分析物的超灵敏检测克服了低浓度，因为这种方法的检测极限远远超过(100倍)具有显色读数的传统方法(例如，酶联免疫吸附试验)。通过将超灵敏检测与多路复用相结合，可以在一小份珍贵样品中测量许多生物分析物，从而提供了一种解决方案。我们阿尔伯塔大学的研究团队非常需要灵敏的测量。麦考利实验室研究结合碳水化合物的免疫调节受体，尤其对大脑中的免疫细胞小胶质细胞感兴趣；理查德实验室研究营养对免疫动态平衡的影响；乐于接受的实验室研究微生物在牲畜物种肠道健康中的作用；菲尔德实验室研究出生后饮食中的脂肪在支持免疫细胞发育方面的作用。要在这些领域做出根本性的发现，需要对生物分子进行精确和灵敏的检测，例如：细胞因子、趋化因子、磷酸化信号分子、抗体和代谢物。艾伯塔大学校园里唯一公开提供的电化学发光读取器最近达到了使用寿命的尽头，不再运行，因为没有修复这个仪器的部件。这使得许多研究小组陷入了无法进行这些测量的困境。因此，迫切需要一种安装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以及艾伯塔大学其他几十个研究小组的研究项目至关重要。之前的仪器安装在艾伯塔省糖尿病研究所的分子核心设施中，校园内的许多小组都很好地使用了它，平均每年运行200多个盘子。因此，我们正在申请资金，用于从复杂的生物样本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，获得一个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。

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.