CORE--ELISA MICROARRAY FACILITY

增加的氧化应激和炎症反应是暴露于环境中的常见效应。 剂.有许多推定的蛋白质标记的环境损害，虽然有有限的， 了解这些蛋白质对监测人类低剂量接触的有用性。除了 假定的蛋白质标记物，项目1和2中提出的蛋白质组学和DNA微阵列分析， 预计将确定新的候选生物标志物暴露于侧流烟雾。这样做的目的 ELISA微阵列的核心是评估候选蛋白质生物标志物的有用性， 环境暴露。对于项目1，我们将在约120个人血浆样品中筛选约50种蛋白质， 识别对香烟烟雾作出反应而改变的蛋白质。虽然大多数分析都是 使用商业试剂进行，该核心还将开发用于多达10种生物标志物的夹心ELISA 商业测定法对此是不可用的。对于项目2，使用了一个亚组（~ 20）的蛋白质筛选， 将在暴露于香烟烟雾、脂多糖、CCl 4或百草枯的小鼠中分析人类。的 小鼠数据将鉴定显示对香烟烟雾暴露的选择性的标记。已知的数据 标记物也将提供基线信息，用于与新的生物标记物如硝化的 在U 54计划中鉴定的蛋白质。由于ELISA微阵列形式是一种高效的系统， 通过评估多种亲和试剂，一旦我们确定了暴露的最佳候选生物标志物， 将使用迄今为止发现的最有希望的标记筛选500多个人类血浆样本。以来 将在项目#3中开发的现场多路复用传感器限于4或5个测定，这些ELISA 微阵列测试对于筛选候选生物标志物的数量并由此鉴定 最有用的一组蛋白质此外，由于ELISA分析常规用作免疫分析。 定量临床测试，我们开发的任何ELISA都可以直接转化为标准临床测试。 因此，该核心将促进常规临床测试和用于评估的现场传感器的开发。 暴露于环境刺激，如侧流香烟烟雾。

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.