Targeted Antibody Microarrays:Tool for Toxicoproteomics

描述（由申请人提供）：通过通量DMA微阵列分析基因表达，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。已经开发了具有/不具有蛋白质的预先分离的质谱法，但是实验的负担在用户身上。 在第一阶段，我们成功地开发和商业化的大鼠药物代谢酶抗体微阵列包含76个抗体的细胞色素P450和辅酶，第二阶段药物代谢酶，骨化相关蛋白，管家蛋白和Flag内部控制蛋白。 在第二阶段，我们将（1）进一步改进大鼠聚丙烯酰胺基水凝胶阵列，（2）改进和开发用各种表面材料涂覆的芯片的额外阵列，（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）探索抗体微阵列的实用性并发现5类毒物的标记物。 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是毒理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.