Targeted Antibody Microarrays:Tool for Toxicoproteomics

描述(申请人提供)：基因表达已经通过吞吐量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，由于基因的翻译/降解速率和翻译后修饰的不同，基因的mRNA表达可能与蛋白质表达不相关。有或没有预先分离蛋白质的质谱学已经开发出来，但实验的负担落在了使用者身上。 在第一阶段，我们成功地开发并商业化了大鼠药物代谢酶抗体微阵列，其中包含76种抗体，包括细胞色素P450及其辅酶、第二阶段药物代谢酶、细胞凋亡相关蛋白、看家蛋白和Flag内部对照蛋白的抗体。 在第二阶段，我们将(1)进一步改进基于聚丙烯酰胺的大鼠水凝胶阵列，(2)改进和开发额外的带有各种表面材料的芯片阵列，(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)开发人药物代谢酶抗体微阵列，(6)探索抗体阵列的用途，发现5类毒物的标志物。 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。

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.