New microscale sample processing approaches for application to proteomics and bioanalysis

适用于蛋白质组学和生物分析的新型微量样品处理方法

• 批准号: RGPIN-2016-06636
• 负责人: Waldron, Karen
• 金额: $ 1.82万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=667971
• 关键词: New; microscale; sample; processing; approaches

The health and the well-being of Canadians rely not only on accurate diagnosis and effective treatment but also a pollution-free environment in which to live. Chemical analysis plays a key role in such goals by helping uncover the causes of disease, or by following a patient's drug therapy response, or by testing water, soil and air for toxins to both plants and animals. By far the most common methods for said chemical analyses are grouped under the term “analytical separations”, encompassing the powerful techniques of chromatography, electrophoresis and mass spectrometry. The complexity of samples submitted for chemical analysis, sometimes containing over a thousand different compounds, is such that rigorous sample preparation techniques are required prior to the analytical separation. In fact, the selectivity and detection sensitivity of the above-mentioned analytical separation methods improve greatly with preparation techniques tailored to the analysis. Therefore, our research program is dedicated to developing and optimizing sample preparation methods with specific application to microliter scale bioanalysis and towards enhancing the power of state-of-the-art analytical separations. ***The identification of proteins and the understanding of their biological interactions is key to developing diagnostic and therapeutic strategies. Therefore, we are designing sample preparation methods that aid in protein determination, including proteins at low abundance because these are the ones suspected to have relevance for disease diagnosis. We are developing fractionation strategies and enzymatic methods that render proteins easier to analyze by mass spectrometry and address the current trend towards miniaturization. Capillary electrophoresis is our main microscale method, which is ideally suited because nanolitre volumes can be injected, it is easily automated and it can be translated into microchip format for additional miniaturization. Our immobilized enzyme microreactor methods are being developed to allow simple construction and rapid protein digestion for better mass spectrometry analysis. Innovative combinations of these microscale techniques and methods applied to minute sample volumes has the long term potential to impact disease diagnosis and monitoring in humans and animals. **

加拿大人的健康和福祉不仅依赖于准确的诊断和有效的治疗，而且还依赖于无污染的生活环境。化学分析在这些目标中发挥着关键作用，有助于发现疾病的原因，或通过跟踪患者的药物治疗反应，或通过测试水，土壤和空气中的植物和动物毒素。到目前为止，用于所述化学分析的最常见的方法被归类在术语“分析分离”下，包括色谱法、电泳法和质谱法的强大技术。提交用于化学分析的样品的复杂性，有时包含超过一千种不同的化合物，使得在分析分离之前需要严格的样品制备技术。事实上，上述分析分离方法的选择性和检测灵敏度随着针对分析定制的制备技术而大大提高。因此，我们的研究计划致力于开发和优化样品制备方法，具体应用于微升规模的生物分析，并提高最先进的分析分离的能力。* 蛋白质的鉴定和对它们的生物学相互作用的理解是开发诊断和治疗策略的关键。因此，我们正在设计有助于蛋白质测定的样品制备方法，包括低丰度蛋白质，因为这些蛋白质被怀疑与疾病诊断相关。我们正在开发分馏策略和酶促方法，使蛋白质更容易通过质谱分析，并解决目前的小型化趋势。毛细管电泳是我们主要的微尺度方法，这是非常适合的，因为可以注入纳升体积，它很容易自动化，它可以转换成微芯片格式，以进一步微型化。我们的固定化酶微反应器方法正在开发中，以允许简单的结构和快速的蛋白质消化更好的质谱分析。这些微尺度技术和方法的创新组合应用于微小的样品体积，具有影响人类和动物疾病诊断和监测的长期潜力。**