Development and Implementation of Immunoassays

免疫分析的开发和实施

• 批准号: 6900539
• 负责人: Shirley Jacqueline Gee
• 金额: $ 26.58万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6900539
• 关键词: acrylamides; biohazard detection; biomarker; biphenyl compounds; ciprofloxacin; environmental contamination; environmental exposure; human tissue; immunologic assay /test; insecticide resistance; laboratory rabbit; pyrethroid; technology /technique development; urinalysis

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.

该提案的总体目的是快速的发展，验证和实施 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暴露的生物标志物。要了解化学暴露对人类种群的影响，必须量化环境水平以及化学物质的内部剂量。将暴露与作用联系起来的研究需要分析各种矩阵中的大量样品（即土壤，沉积物，水，空气，尿液，血液）。免疫测定有一个 在临床领域的敏感性，速度和成本效益在临床领域表现出声誉。这些优势使其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。一个项目的目的是完成拟除虫菊酯杀虫剂代谢产物的测定。测定库将经过验证，然后使用暴露的尿液样本实施 人口。另一个目的是为生物标志物和环境分析开发新的化合物和类型测定法。所选的化合物是危险化合物聚溴二苯基醚，丙烯酰胺和非氯烷基聚氯化双苯基，杀虫剂菲普罗尼尔和新出现的污染物抗生素抗生素环丙沙星。该复合列表反映了紧迫的环境/人类健康问题以及其他超级基金的需求 程序中的项目。这些测定将格式化用于单层和多体分析阵列的快速实验室分析以及用于现场使用。在改进测定技术方面将付出巨大的努力。一种方法是通过消除基质效应来提高灵敏度。为此，将评估各种萃取方法和清理方法（即免疫症状，固相微萃取，微波辅助提取），以便于具有免疫测定的界面，提高敏感性，鲁棒性，鲁棒性和分析速度。推进与免疫诊断的技术以及对新生物传感器设计的应用和开发相关的技术也是一个目标。改善用作标签和酶底物的酶以及新型荧光团作为标签的开发是很小的步骤，最终将应用于高通量生物传感器的发展。