Protease chain reactions for molecular analysis of cancer markers

用于癌症标志物分子分析的蛋白酶链反应

基本信息

批准号：
8896302
负责人：
Biao Ruan
金额：
$ 7.35万
依托单位：
POTOMAC AFFINITY PROTEINS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-19 至 2016-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Our objective is to develop a Protease Chain Reaction technology (ProCR) that will enable ultra- sensitive and highly-quantitative detection of cancer markers. The basic element of a protease chain reaction is a protease-inhibitor complex. The protease is inactive when bound to the inhibitor but, once freed, is capable of cleaving the inhibitor and releasing additional free protease. A trace of free protease initiates the chain reaction and eventually causes the release of all protease from the inhibitory complex. The lag time preceding the full release of protease is determined by the initial concentration of free protease. The Phase I project developed a first-generation of self-amplifying complexes that form the core of the ProCR detection technology. All Phase I milestones were significantly exceeded. In Phase II we will develop powerful molecular sensors based on self- amplifying complexes. A sensor consists of three elements: 1) a sensing element that responds to the presence of an analyte; 2) the processing/computational element that amplifies and quantifies the signal from the detection element; and 3) the transducer element that produces an optical signal. Through this combination, extremely sophisticated enzymatic sensors will be built to detect cancer markers. Accordingly the three experimental Aims are to: 1) Engineer sensing elements; 2) Engineer enhanced processing/computational elements; and 3) Engineer signaling components. Fundamentally, a protein chain reaction is a powerful analogue computer with two key characteristics that greatly facilitate the detection of target molecules. 1) It can convert th concentration of a specific target molecule into a time signature. 2) It can create enormous signal amplification, analogous to the amplification of DNA by a polymerase chain reaction (PCR). Thus detection is enabled because the final observable signal produced by a target molecule can be very large and the time lag until onset of the signal is precisely correlated with the concentration of target molecule. The long term goal is to develop protease-inhibitor complexes as enzymatic nano-processors which can be combined to detect multiple signals and to control output with multiple logic gates.

描述（由申请人提供）：我们的目的是开发一种蛋白酶链反应技术（PRACR），该技术将对癌症标志物进行超敏感和高度量化的检测。蛋白酶链反应的基本元素是蛋白酶抑制剂复合物。蛋白酶在与抑制剂结合时无活性，但是一旦释放，蛋白酶就能裂解抑制剂并释放额外的游离蛋白酶。自由蛋白酶的痕迹会引起链反应，并最终导致所有蛋白酶从抑制性络合物中释放出来。蛋白酶的完整释放之前的滞后时间取决于游离蛋白酶的初始浓度。第一阶段项目开发了构成PROCR检测技术核心的自我扩增复合物的第一代。所有第一阶段的里程碑都大大超过了。在第二阶段，我们将基于自我扩增的复合物开发强大的分子传感器。传感器由三个要素组成：1）对分析物的存在响应的传感元素； 2）放大和量化检测元件的信号的处理/计算元素； 3）产生光学信号的传感器元件。通过这种组合，将构建极其复杂的酶传感器来检测癌症标志物。因此，三个实验目的是：1）工程师感应元素； 2）工程师增强的处理/计算元素； 3）工程师信令组件。从根本上讲，蛋白质链反应是一款强大的模拟计算机，具有两个关键特征，极大地促进了靶分子的检测。 1）它可以将特定目标分子的TH浓度转换为时间签名。 2）它可以产生巨大的信号扩增，类似于通过聚合酶链反应（PCR）扩增DNA。因此启用了检测，因为目标分子产生的最终可观察信号可能非常大，并且时间滞后直到信号发作与靶分子的浓度完全相关。长期目标是开发蛋白酶抑制剂复合物作为酶促纳米处理器，可以组合以检测多个信号并用多个逻辑门控制输出。