Protein NanoArray for Cytokine Profiling in Mouse Models

描述（由申请人提供）：这是一项I期SBIR提案，旨在测试使用超小型化蛋白质纳米阵列分析小鼠模型系统中细胞因子表达的可行性。BioForce Nanosciences, Inc.已经开发了构建NanoArrays/TM的专有技术，这是一种超小型化的分析平台，允许在单个标准微阵列点占用的相同表面空间内进行1000或更多的分子测试。利用该技术，我们构建了一个9组分抗体三明治NanoArray，并利用小鼠重组细胞因子和细胞培养模型系统测试了其灵敏度、精密度和线性度。与传统的微阵列格式相比，NanoArray平台提供的尺寸减小提供了一种机制，用于创建需要极小样本量的诊断测试，接近单个细胞的样本量。该分析系统特别适合于涉及小动物模型系统的研究，在这些研究中，传统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可靠的蛋白质分析数据，这些数据来自小鼠尾出血、针活检抽吸和人类血液样本的手指刺痛。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读取，允许从单个图像收集数据。这一特性为预算和/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实现的临床测试打开了大门。

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.