Protein NanoArray for Cytokine Profiling in Mouse Models

描述（由申请方提供）：这是一项I期SBIR提案，旨在测试使用超小型蛋白质NanoArray分析小鼠模型系统中细胞因子表达的可行性。BioForce Nanosciences，Inc.，已经开发了用于构建NanoArrays/TM的专有技术，NanoArrays/TM是一种超小型化的测定平台，允许在单个标准微阵列点所占据的相同表面空间中进行一千个或更多的分子测试。利用该技术，我们提出了建立一个9组分抗体夹心NanoArray，并使用小鼠重组细胞因子和细胞培养模型系统测试其灵敏度，精密度和线性。相对于传统的微阵列形式，NanoArray平台提供的尺寸减小提供了一种用于创建诊断测试的机制，该诊断测试需要非常小的样品体积，接近单细胞的体积。该测定系统特别适合于涉及小动物模型系统的研究，其中常规分析技术可能需要致死量的起始样品材料。我们预测，我们的系统将提供强大的蛋白质分析数据，从小鼠尾出血，穿刺活检吸出物，和人类血液样本的手指刺破。此外，NanoArray平台提供了小型化的好处，但仍然允许使用标准荧光方法进行读数，从而允许从单个图像中收集数据。这一属性为预算和/或仪器资源有限的科学家增加了基于阵列的生物分析的可用性，并为现有方法目前无法进行的临床测试打开了大门。 我们已经为我们的I期研究选择了细胞因子生物标志物。细胞因子与多种肾脏和消化系统疾病的发病机制有关，包括糖尿病、进行性肾衰竭和炎症性肠病。使用细胞因子基准将使我们能够产生一个纳米阵列生物标志物筛选，可以与现有的标准ELISA和基于微阵列的测试进行比较，并立即对使用转基因小鼠疾病模型的研究人员有用。该检测方法的开发将为针对其他疾病靶点和/或信号通路定制的蛋白质谱芯片的第11阶段开发铺平道路，用于小鼠模型系统以及涉及极小样本量的人体测试。

DESCRIPTION (provided by the applicant): This is a Phase I SBIR proposal to test the feasibility of using an ultra miniaturized protein NanoArray for profiling cytokine expression in mouse model systems. BioForce Nanosciences, Inc., has developed proprietary technology for the construction of NanoArrays/TM, an ultra miniaturized assay platform that allows a thousand or more molecular tests to be carried out in the same surface space occupied by a single standard microarray spot. Using this technology, we propose to build a 9 component antibody sandwich NanoArray and test its sensitivity, precision and linearity using mouse recombinant cytokines and cell culture model systems. The size reduction offered by the NanoArray platform, relative to the conventional microarray format, provides a mechanism for the creation of diagnostic tests that require extremely small sample volumes, approaching that of a single cell. This assay system is particularly well-suited for studies involving small animal model systems, where conventional analysis techniques may require lethal amounts of starting sample material. We predict that our system will provide robust protein profiling data from mouse tail-bleeds, needle biopsy aspirates, and finger pricks of human blood samples. Furthermore, the NanoArray platform provides the benefits of miniaturization but still allows readout using standard fluorescence methods permitting data collection from a single image. This attribute increases the availability of array-based bioanalyses for scientists with limited budgets and/or instrumentation resources, and opens the doorway for clinical testing not currently possible with existing methodology. We have chosen cytokine biomarkers for our Phase I study. Cytokines have been implicated in the pathogenesis of a wide variety of diseases of the kidney and digestive systems, including diabetes, progressive renal failure and inflammatory bowel disease. Using the cytokine benchmark will allow us to produce a NanoArray biomarker screen that can be compared with existing standard ELISA and microarray-based tests, and be immediately useful to researchers using transgenic mouse models of disease. Development of this assay will pave the way for Phase 11development of protein profiling chips tailored to other disease targets and/or signaling pathways, for use in mouse model systems, and for human testing involving extremely small sample volumes.