Protein NanoArray for Cytokine Profiling in Mouse Models

用于小鼠模型细胞因子分析的蛋白质纳米阵列

• 批准号: 6748544
• 负责人: Eric R HENDERSON
• 金额: $ 25.03万
• 依托单位: BIOFORCE NANOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-20 至 2005-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6748544
• 关键词: animal tissue; biomarker; biomedical equipment development; cytokine; fluorescent dye /probe; laboratory mouse; method development; miniature biomedical equipment; nanotechnology; protein quantitation /detection; protein structure function; tissue /cell culture

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.

描述（由申请人提供）：这是I期SBIR建议，用于测试使用超小化蛋白纳米array在小鼠模型系统中分析细胞因子表达的可行性。 BioForce Nanosciences，Inc。已开发了用于构建纳米阵列/TM的专有技术，纳米阵列/TM是一个超小型化的测定平台，允许在单个标准微阵列占据的同一表面空间中进行一千个或更多的分子测试。使用这项技术，我们建议使用小鼠重组细胞因子和细胞培养模型系统来构建9个组分抗体三明治纳米阵列，并测试其敏感性，精度和线性。相对于传统的微阵列格式，纳米array平台提供的尺寸减小为创建需要极小样品量的诊断测试提供了一种机制，即接近单个单元格。该测定系统特别适合涉及小动物模型系统的研究，在这种研究中，常规分析技术可能需要致命的起始样品材料。我们预测，我们的系统将提供来自小鼠尾巴，针头活检抽吸物和人类血液样本的手指刺的强大蛋白质分析数据。此外，NanoArray平台提供了微型化的好处，但仍然允许使用标准荧光方法进行读数，从而可以从单个图像中收集数据。该属性增加了预算和/或仪器资源有限的科学家的基于阵列的生物分析的可用性，并为目前使用现有方法开放了临床测试的门口。 我们在我们的I期研究中选择了细胞因子生物标志物。细胞因子与肾脏和消化系统多种疾病的发病机理有关，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生成可以将其与现有标准ELISA和基于微阵列的测试进行比较的纳米阵列生物标志物屏幕，并立即使用转基因小鼠疾病模型来对研究人员有用。该测定法的开发将为第11阶段开发的蛋白质谱分析芯片铺平道路，适合其他疾病靶标和/或信号通路，用于小鼠模型系统以及涉及极小样品体积的人体测试。