Targeted Antibody Microarrays:Tool for Toxicoproteomics

靶向抗体微阵列：毒理学蛋白质组学工具

• 批准号: 6938769
• 负责人: Hyesook Kim
• 金额: $ 46.26万
• 依托单位: DETROIT R & D, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6938769
• 关键词: SDS polyacrylamide gel electrophoresis; antibody specificity; biological signal transduction; biomarker; drug metabolism; environmental toxicology; enzyme activity; gel; gene expression; laboratory mouse; laboratory rat; method development; microarray technology; polyacrylamide; proteomics; surface coating; toxicant screening; toxin metabolism; western blottings

DESCRIPTION (provided by applicant): Gene expression has been analyzed by throughput DMA microarrays which simultaneously monitor the expression of thousands of genes. However, mRNA expression of a gene may not correlate with protein expression because of the differences in rates of mRNA translation/degradation and post-translational modification. Mass spectrometry with/without prior separation of proteins has been developed but the burden of the experiment is on the user. During Phase I, we successfully developed and commercialized rat drug-metabolizing enzyme antibody microarrays containing 76 antibodies for cytochromes P450 and co-enzymes, phase II drug-metabolizing enzymes, apoptosis-related proteins, house-keeping proteins and Flag internal control protein. During Phase II, we will (1) further improve rat polyacrylamide-based hydrogel arrays, (2) improve and develop additional arrays with chips coated with various surface materials, (3) develop and explore signal enhancing methods, (4) develop mouse drug-metabolizing enzyme antibody microarrays, (5) develop human drug-metabolizing enzyme antibody microarrays and (6) explore utility of antibody arrays and discover markers for 5 classes of toxicants. The targeted arrays produced with form-specific antibodies of closely related proteins involved with drug and environmental toxicant metabolism will be an invaluable tool for toxicoproteomics.

描述（由申请人提供）：基因表达已通过吞吐量 DMA 微阵列进行分析，该微阵列同时监测数千个基因的表达。然而，由于 mRNA 翻译/降解和翻译后修饰速率的差异，基因的 mRNA 表达可能与蛋白质表达不相关。已经开发出预先分离蛋白质或不分离蛋白质的质谱法，但实验的负担由用户承担。 一期阶段，我们成功开发并商业化了含有76种细胞色素P450和辅酶抗体的大鼠药物代谢酶抗体微阵列、二期药物代谢酶、凋亡相关蛋白、管家蛋白和Flag内参蛋白。 在第二阶段，我们将（1）进一步改进基于大鼠聚丙烯酰胺的水凝胶阵列，（2）改进和开发带有涂有各种表面材料的芯片的其他阵列，（3）开发和探索信号增强方法，（4）开发小鼠药物-代谢酶抗体微阵列，(5)开发人类药物代谢酶抗体微阵列，(6)探索抗体阵列的实用性并发现5类毒物的标记物。 用与药物和环境毒物代谢密切相关的蛋白质的形式特异性抗体产生的靶向阵列将成为毒物蛋白质组学的宝贵工具。