Protein NanoArray for Cytokine Profiling in Mouse Models

描述(由申请人提供)：这是一项第一阶段的SBIR提案，旨在测试在小鼠模型系统中使用超微型蛋白质纳米阵列来分析细胞因子表达的可行性。Bioforce Nanosciences，Inc.开发了用于构建纳米阵列/TM的专利技术，这是一个超小型分析平台，允许在单个标准微阵列点占据的相同表面空间内进行1000或更多的分子测试。利用这一技术，我们构建了9组分抗体夹心纳米阵列，并利用小鼠重组细胞因子和细胞培养模型系统对其灵敏度、精密度和线性进行了测试。与传统微阵列格式相比，纳米阵列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，该诊断测试需要极小的样本体积，接近单个细胞的体积。这种分析系统特别适合于涉及小型动物模型系统的研究，在这些系统中，传统的分析技术可能需要致命数量的起始样品材料。我们预测，我们的系统将提供强大的蛋白质图谱数据，从小鼠尾巴出血，针刺活组织抽吸，和指刺的人类血液样本。此外，纳米阵列平台提供了小型化的好处，但仍然允许使用允许从单个图像收集数据的标准荧光方法进行读出。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，并为临床测试打开了大门，这是现有方法学目前无法实现的。 我们已经为我们的第一阶段研究选择了细胞因子生物标志物。细胞因子参与了多种肾脏和消化系统疾病的发病机制，包括糖尿病、进行性肾功能衰竭和炎症性肠病。使用细胞因子基准将使我们能够生产出一种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。这项测试的开发将为第11阶段开发蛋白质图谱芯片铺平道路，这些芯片是为其他疾病靶点和/或信号通路量身定做的，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.