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Elements and systems for ultrasensitive protein analysis

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:
依托单位国家:
加拿大
项目类别:
Discovery Grants Program - Individual
财政年份:
2016
资助国家:
加拿大
项目状态:
已结题
起止时间:
2016-01-01 至 2017-12-31

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中文摘要
翻译
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。近年来出现了超灵敏的蛋白质分析技术,但这些技术复杂,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,我们建议开发新的技术,用于超灵敏的,多重的蛋白质分析,用于生物标记物的发现和护理点测试。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。(I)和(Ii)在设计上都考虑到了多路复用,并且都被设计为抑制交叉反应。(Iii)我们将开发3D打印毛细管微流控电路,这种电路是根据类似电子电路的元素库设计的,可以在几个小时内通过快速原型制作而成。这些毛细管微流体是自供电和可编程的,将被开发为低成本、用于实验室和护理点分析的超灵敏测试。 这一计划将对检测科学和技术产生重大影响,并造福于人类健康。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。
英文摘要
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) & (ii) are designed with multiplexing in mind and are designed to suppress cross-reactivity. (iii) We will develop 3D printed capillary microfluidic circuits that are designed from a library of elements, akin to electronic circuits, and can be made within a few hours by rapid prototyping. These capillary microfluidics are self-powered and programmable, and will be developed into low cost, ultrasensitive test for laboratory and point-of-care analysis. This program is poised to significantly impact assay 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.
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Integrative and Translational Biomedical Engineering
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    2022
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Microfluidic Chain Reaction: Exploration, Expansion, and Application
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  • 批准号:
    CRC-2018-00085
  • 项目类别:
    Canada Research Chairs
  • 资助金额:
    $14.57万
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    2021
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Elements and systems for ultrasensitive protein analysis
  • 批准号:
    RGPIN-2016-06723
  • 项目类别:
    Discovery Grants Program - Individual
  • 资助金额:
    $7.87万
  • 财政年份:
    2021
  • 负责人:
    Juncker, David
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