Elements and systems for ultrasensitive protein analysis

决定许多疾病（特别是癌症）结局的最重要因素是诊断时的疾病进展。不幸的是，疾病通常只在症状出现后的晚期才被发现。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，并可用作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标，就有可能早期筛查和诊断疾病。目前临床上使用的血液测试检测到的蛋白质仅占血液蛋白质的100亿分之一。然而，为了发现用于早期诊断的生物标志物，需要能够同时测量多种蛋白质并且灵敏度高出一千到一百万倍的技术。最近出现了超灵敏蛋白质分析技术，但它们很复杂，只能在实验室中操作，多重蛋白质测量要么不可行，要么只能以牺牲灵敏度和可靠性为代价，因为测定设计没有考虑多种试剂之间的交叉反应性。 在这里，我们建议开发用于生物标志物发现和即时检测的蛋白质超灵敏多重分析的新技术。这项工作将沿着沿着三个轴组织：（i）一个数字纳米点阵列，包括数百万个纳米尺度的点，每个点可以捕获蛋白质将使用我们开发的低成本图案化方法。捕获的蛋白质将以单分子灵敏度检测并计数。我们介绍了一种新的噪声抑制算法的基础上空间discromination，可以提高检测灵敏度和鲁棒性。(ii)将开发一种新的基于微珠的共定位固定化试剂的测定法。珠子将被添加到样品中，并混合，从而“舀”起所有的蛋白质，然后将被检测到。科学和技术，造福人类健康。这笔赠款还将提供5个本科生和研究生的跨学科环境，包括纳米技术，医疗保健和商业化的培训年度支持，并培养明天的科学家，企业家和领导者。

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.