Development and Implementation of Immunoassays

本提案的总体目标是开发、验证和实施快速 基于免疫化学的方法，用于分析危险材料及其分解产物，作为人类接触的生物标志物。为了了解化学品接触对人类的影响，必须对化学品的环境水平和体内剂量进行量化。将接触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。免疫分析具有 在临床竞技场中因其灵敏度、速度和成本效益而表现出声誉。这些优点使其成为测量尿液代谢物作为暴露生物标志物的理想选择。此外，该技术已被应用于环境分析。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分析开发。将对测定库进行验证，然后使用来自暴露者的尿样实施 人口另一个目标是开发新的化合物和类别选择性测定，用于生物标志物和环境分析。选定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、杀虫剂氟虫腈和新出现的污染物抗生素环丙沙星。这份综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。该检测试剂盒将被格式化，用于单分析物和多分析物阵列的快速实验室分析以及现场使用。将在改进分析技术方面作出相当大的努力。一种方法是通过消除基质效应来提高灵敏度。为此，将评价各种提取和净化方法（即免疫亲和法、固相微萃取、微波辅助提取），以便于与免疫测定结合，提高灵敏度、稳健性和分析速度。推进与免疫诊断相关的技术以及新生物传感器设计的应用和开发也是一个目标。对用作标记的酶和酶底物的改进以及作为标记的新型荧光团的开发是最终将应用于高通量生物传感器的开发的小步骤。

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.