Development & Implementation of Immunoassays for Human & Environmental Monitoring

该提案的总体目标是开发、验证和实施快速 作为人体暴露生物标志物的危险物质及其分解产物的免疫化学分析方法。为了了解化学品暴露对人类人口的影响，必须对环境水平以及化学品的内部剂量进行量化。将暴露与影响联系起来的研究需要分析各种基质(即土壤、沉积物、水、空气、尿液、血液)中的大量样本。免疫分析有一种 在临床领域以其敏感度、速度和成本效益而享有盛誉。这些优点使它们成为测量尿代谢物作为暴露生物标志物的理想选择。此外，该技术已应用于环境分析。其中一个项目的目标是完成拟除虫菊酯类杀虫剂代谢物的检测开发。化验库将进行验证，然后使用暴露的尿样进行实施 人口。另一个目标是开发新的化合物和类别选择性分析方法，用于生物标记物和环境分析。选择的化合物是危险化合物多溴联苯醚、丙烯酰胺和非共面多氯联苯、杀虫剂氟虫腈和新兴的污染抗生素环丙沙星。这份复合清单反映了紧迫的环境/人类健康问题以及其他超级基金的需求 计划内的项目。分析的格式将在单一和多个分析物阵列中进行快速实验室分析，并供现场使用。将在改进检测技术方面做出相当大的努力。一种方法是通过消除基质效应来提高灵敏度。为此，将评估各种提取和净化方法(即免疫亲和、固相微萃取、微波辅助提取)，以便于与免疫分析进行接口，提高分析的灵敏度、稳健性和速度。推进与免疫诊断相关的技术以及应用和开发新的生物传感器设计也是一个目标。对用作标记和酶底物的酶的改进以及作为标记的新型荧光团的开发都是很小的步骤，最终将应用于高通量生物传感器的开发。

The overall objective of this proposal is the development, validation and implementation of rapid immunochemical-based methods for the analysis of hazardous materials and their breakdown products as biomarkers of human exposure. To understand the impact of chemical exposures on human populations, environmental levels as well as the internal doses of the chemicals must be quantified. Studies that link exposure to effect require analysis of large numbers of samples in a variety of matrices (i.e. soil, sediment, water, air, urine, blood). Immunoassays have a demonstrated reputation in the clinical arena for their sensitivity, speed, and cost effectiveness. These advantages make them ideal for measuring urinary metabolites as biomarkers of exposure. In addition, the technique has been applied to environmental analysis. One project aim is to complete the assay development of metabolites of pyrethroid insecticides. The library of assays will be validated and then implemented using urine samples from an exposed population. Another aim is to develop new compound and class-selective assays for both biomarker and environmental analysis. The compounds selected are the hazardous compounds polybrominated diphenyl ethers, acrylamide, and noncoplanar poly chlorinated biphenyls, the insecticide fipronil and the emerging pollutant antibiotic ciprofloxacin. This compound list reflects pressing environmental/human health problems as well as the needs of other Superfund projects within the Program. The assays will be formatted for rapid laboratory analysis in single and multianalyte arrays and for field use. Considerable effort will be placed on improving assay technology. One approach is to improve sensitivity by eliminating matrix effects. Toward this end a variety of extractions and cleanup methods will be evaluated (i.e. immunoaffmity, solid phase microextraction, microwave-assisted extraction) for ease of interface with immunoassays, improvements to sensitivity, robustness and speed of analysis. Advancing the technologies associated with immunodiagnostics and application to and development of new biosensor designs is also a goal. Improvements to enzymes used as labels and to enzyme substrates as well as development of novel fluorophores as labels are small steps that will be applied ultimately to the development of high-throughput biosensors.