CORE--ELISA MICROARRAY FACILITY

核心--ELISA 微阵列设备

• 批准号: 7637343
• 负责人: RICHARD C ZANGAR
• 金额: $ 11.55万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-10 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7637343
• 关键词: Affinity; Antibodies; Antigens; Biological Assay; Biological Markers; Clinical; DNA Microarray Chip; DNA Microarray format; Data; Dose; Environmental Exposure; Enzyme-Linked Immunosorbent Assay; Exposure to; Human; Inflammation; Inflammatory Response; Knowledge; Lipopolysaccharides; Monitor; Mus; Nitrates; Numbers; Oxidative Stress; Paraquat; Plasma; Proteins; Proteomics; Purpose; Reagent; Sampling; Screening procedure; Set protein; Side; Site; Smoke; Smoker; Source; Standards of Weights and Measures; Stimulus; Stream; System; Testing; Toxic effect; Translating; cigarette smoking; environmental agent; environmental stressor; instrument; nitrate; novel; programs; research clinical testing; response; sensor

Increased oxidative stress and inflammatory responses are common effects of exposure to environmental agents. There are many putative protein markers of environmental insult, although there is limited knowledge of the usefulness of these proteins for monitoring low-dose exposures in humans. In addition to putative protein markers, the proteomic and DNA microarray analyses proposed in Projects 1 and 2 are expected to identify new candidate biomarkers for exposure to side-stream smoke. The purpose of this ELISA microarray core is to evaluate the usefulness of candidate protein biomarkers for assessing environmental exposures. For Project 1, we will screen -50 proteins in -120 human plasma samples to identify proteins that are altered in response to cigarette smoke. Although most of these analyses will be undertaken using commercial reagents, this core will also develop sandwich ELISA's for up to 10 biomarkers for which commercial assays are unavailable. For Project 2, a subset (-20) of the proteins screened in humans will be analyzed in mice exposed to cigarette smoke, lipopolysaccharide, CCI4 or paraquat. The mouse data will identify markers that show selectivity for cigarette smoke exposure. The data for known markers will also provide baseline information for comparison with novel biomarkers such as nitrated proteins identified in this U54 program. Since the ELISA microarray format is a highly efficient system for evaluating multiple affinity reagents, once we have identified the best candidate biomarkers of exposure, we will screen over 500 human plasma samples using the most promising markers identified so far. Since the on-site multiplexed sensor that will be developed in Project #3 is limited to 4 or 5 assays, these ELISA microarray tests are essential to winnow down the number of candidate biomarkers and thereby identify the most useful set of proteins for this instrument. Furthermore, since ELISA analyses are routinely used as a quantitative clinical test, any ELISA's we develop can be directly translated to a standard clinical test. Therefore, this core will facilitate the develop of routine clinical tests and of a on-site sensor for evaluating exposure to environmental stimuli such as side-stream cigarette smoke.

氧化应激和炎症反应增加是暴露在环境中的常见后果 探员们。有许多假定的环境侮辱蛋白质标记物，尽管有限 了解这些蛋白质在监测人体低剂量暴露中的作用。除了……之外 假定的蛋白质标记，项目1和2中提出的蛋白质组和DNA微阵列分析是 预计将确定暴露在侧流烟雾中的新的候选生物标志物。这样做的目的是 ELISA微阵列的核心是评估候选蛋白质生物标志物的有用性 暴露在环境中。对于项目1，我们将在-120个人体血浆样本中筛选-50个蛋白质，以 识别因吸烟而改变的蛋白质。尽管这些分析中的大多数都将是 使用商业试剂，该核心还将开发最多10个生物标志物的夹心ELISAs 无法进行商业检测的产品。对于项目2，筛选到的蛋白质子集(-20) 人类将在暴露于香烟烟雾、脂多糖、CCI4或百草枯的小鼠身上进行分析。这个 鼠标数据将识别显示香烟烟雾暴露的选择性的标记。已知的数据 标记物还将提供基线信息，用于与硝化等新型生物标记物进行比较 在这个U54计划中确定的蛋白质。由于ELISA微阵列格式是一种高效的系统 评估多种亲和试剂，一旦我们确定了暴露的最佳候选生物标记物，我们 将使用到目前为止发现的最有希望的标记物来筛选500多个人类血浆样本。自.以来 将在项目#3中开发的现场多路传感器仅限于4到5次检测，这些酶联免疫吸附试验 微阵列测试对于筛选候选生物标志物的数量至关重要，从而识别 对这台仪器最有用的一组蛋白质。此外，由于酶联免疫分析通常被用作 定量临床试验，我们开发的任何一种ELISA都可以直接转化为标准的临床试验。 因此，这一核心将促进常规临床测试和现场评估传感器的开发。 暴露在环境刺激下，如侧流香烟烟雾。