Elements and systems for ultrasensitive protein analysis

用于超灵敏蛋白质分析的元件和系统

• 批准号: RGPIN-2016-06723
• 负责人: Juncker, David
• 金额: $ 7.87万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=692929
• 关键词: Elements; systems; ultrasensitive; protein; analysis

The single most significant factor determining the outcome of many diseases, and notably cancer, is disease progression at diagnosis. Unfortunately, diseases are typically only detected at an advanced stage, after symptoms appear. A hallmark of diseases is the dysregulation of proteins in cells, and which are released into the blood, and can be used as biomarkers for disease diagnosis. It follows that it might be possible to screen and diagnose diseases early if protein indicators of disease can be discovered. Current blood tests used in the clinic detect a protein constituting as little as one part in 10 billion of blood proteins. However, to discover biomarkers for early diagnosis, technologies that can measure multiple proteins simultaneously and are a thousand to a million times more sensitive are needed. Ultrasensitive protein analysis technologies have emerged recently, but they are complicated, can only operate in the lab, and multiplex protein measurement is either not feasible, or only possible at the expense of sensitivity and reliability because the assay design did not take into consideration cross-reactivity among multiple reagents.****Here, we propose to develop new technologies for ultrasensitive, multiplex analysis of proteins for biomarker discovery and for point-of-care testing. The work will be organized along three axes: (i) A digital nanodot array that comprise millions of nanoscale-dots that each can capture proteins will be made using a low-cost patterning method we developed. Captured proteins will be detected with single-molecule sensitivity, and counted. We introduce a novel noise suppression algorithm based on spatial discromination that can enhance assay sensitivity and robustness. (ii) A novel microbead-based assay with colocalized immobilization of reagents will be developed. Beads will be added to the sample, and mixed, thus “scooping” up all the proteins, which will then be detected. Both (i) ssay science and technology, and benefit human health. This grant will also provide annual support for training of 5 undergraduate and graduate students within a transdisciplinary environment encompassing nanotechnology, healthcare and commercialization, and train tomorrow's scientists, entrepreneurs, and leaders. ********

决定许多疾病(尤其是癌症)结局的最重要的一个因素是确诊时的疾病进展。不幸的是，疾病通常只有在症状出现后的晚期才能被发现。疾病的一个标志是细胞中蛋白质的失调，这些蛋白质被释放到血液中，可以作为疾病诊断的生物标志物。因此，如果能够发现疾病的蛋白质指示物，就有可能及早筛查和诊断疾病。目前用于临床的血液测试检测到一种蛋白质，其组成比例低至100亿血液蛋白质中的一部分。然而，为了发现早期诊断的生物标记物，需要能够同时测量多个蛋白质并且灵敏度高出一千到一百万倍的技术。近年来出现了一些超灵敏的蛋白质分析技术，但这些技术很复杂，只能在实验室中操作，而且由于分析设计没有考虑多个试剂之间的交叉反应，所以多重蛋白质测量要么是不可行的，要么是以牺牲灵敏度和可靠性为代价的。*在这里，我们建议开发新的超灵敏蛋白质多重分析技术，用于生物标志物的发现和护理点检测。这项工作将沿着三个轴组织：(I)使用我们开发的低成本图案化方法制作由数百万个纳米级的点组成的数字纳米点阵列，每个纳米点都可以捕获蛋白质。捕获的蛋白质将以单分子灵敏度进行检测和计数。提出了一种新的基于空间判定的噪声抑制算法，提高了检测的灵敏度和稳健性。(Ii)将开发一种新的基于微珠的试剂共定位固定化分析方法。珠子将被添加到样本中，并混合在一起，这样就可以“挖出”所有的蛋白质，然后被检测出来。既讲科学技术，又有益于人类健康。这笔赠款还将提供年度支持，在包括纳米技术、医疗保健和商业化的跨学科环境中培训5名本科生和研究生，并培训未来的科学家、企业家和领导者。********