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: 6643998
• 负责人: Eric R HENDERSON
• 金额: $ 24.94万
• 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 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6643998
• 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 提案，旨在测试使用超微型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. 开发了用于构建 NanoArrays/TM 的专有技术，这是一个超小型化分析平台，允许在单个标准微阵列点占据的同一表面空间中进行一千个或更多的分子测试。利用该技术，我们建议构建 9 组分抗体夹心纳米阵列，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、精确度和线性。相对于传统的微阵列格式，NanoArray 平台提供的尺寸减小提供了一种创建诊断测试的机制，该诊断测试需要极小的样本量，接近单细胞的样本量。该测定系统特别适合涉及小动物模型系统的研究，其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来自小鼠尾部放血、针活检抽吸物和人类血液样本的手指刺破的可靠蛋白质分析数据。此外，NanoArray 平台提供了小型化的优势，但仍然允许使用标准荧光方法进行读出，从而允许从单个图像收集数据。这一属性提高了预算和/或仪器资源有限的科学家进行基于阵列的生物分析的可用性，并为目前现有方法无法实现的临床测试打开了大门。 我们为我们的第一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生产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标和/或信号通路定制的蛋白质分析芯片的第 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。