Nanoliter-scale HTS and chemogenomics instrumentation

描述(由申请人提供)：拟议项目的总体目标是开发一种纳升规模的液滴仪器，用于高通量筛选(HTS)和化学基因组学。许多生化分析已经在这个平台上进行了演示。我们预计，在第二阶段，将创造一个综合的台式仪器，其吞吐量能够超过10,000,000个数据点/天。在这种筛选中使用的化合物和靶标的数量将比目前使用的方法低1000倍。因此，化合物文库可以针对一系列化学、结构或功能相关的目标进行筛选，甚至可以针对用户可用的所有目标进行筛选，而不必重新合成它。它将在每个反应中使用几纳升试剂溶液进行分析，从而能够快速、经济地针对大型靶板对大型化合物文库进行化学基因组学筛选。为了实现减少试剂消耗和增加产量的优点，试剂必须以一种消除试剂浪费的格式存储，并允许快速、方便地装载。大多数生物化学物质需要低温储存，而且多次冻融循环会降低试剂质量。在第一阶段，我们打算证明，大的化合物文库确实可以存储在与我们的芯片实验室技术兼容的纳升等分格式，然后用于分析。证明使用低温保存的纳升等量试剂进行分析的可行性，将允许在第二阶段以前所未有的吞吐量和经济性开发用于高温超导和化学基因组学的完整纳升仪器系统。

DESCRIPTION (provided by applicant): The overall goal of the proposed project is to develop a nanoliter-scale, droplet-based instrumentation for high-throughput screening (HTS) and chemical genomics. A number of biochemical assays have been demonstrated on this platform. We anticipate creating, in Phase II, an integrated benchtop instrument capable of throughputs exceeding 10,000,000 data points/day. The amounts of both compounds and targets used in such screening will be up to 1,000 times lower than in the currently used methods. Therefore, a compound library can be screened against a family of chemically, structurally or functionally related targets, - or indeed all targets available to a user, - without having to re-synthesize it. It will perform the analyses using several nanoliters of reagent solutions per reaction, thus enabling chemical genomics screening of large compound libraries against large panels of targets rapidly and economically. To realize the advantages of reduced reagent consumption and increased throughput, reagents have to be stored in a format eliminating reagent waste and allowing rapid, convenient loading. Most biochemicals require cryogenic storage, and multiple freeze-thaw cycles reduce reagent quality. In Phase I, we intend to demonstrate that large libraries of compounds can indeed be stored in nanoliter aliquot format compatible with our lab-on-chip technology, and then used in assays. Demonstrating feasibility of assays with cryogenically preserved nanoliter aliquots of reagents will allow development, in Phase II, of a complete nanoliter instrumentation system for HTS and chemical genomics with unprecedented throughput and economy.