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米级、基于液滴的高通量筛选（HTS）和化学基因组学仪器。在这个平台上已经演示了许多生化分析。我们预计在第二阶段创建一个集成的台式仪器，其吞吐量超过1000万个数据点/天。在这种筛选中使用的化合物和目标的数量将比目前使用的方法低1000倍。因此，化合物库可以针对一系列化学上、结构上或功能上相关的靶标进行筛选，或者实际上对用户可用的所有靶标进行筛选，而不必重新合成它。它将在每个反应中使用几纳升的试剂溶液进行分析，从而实现针对大量目标快速经济地进行大型化合物文库的化学基因组筛选。为了实现减少试剂消耗和提高吞吐量的优势，试剂必须以消除试剂浪费和允许快速，方便装载的格式存储。大多数生化试剂需要低温储存，多次冻融循环会降低试剂质量。在第一阶段，我们打算证明大型化合物库确实可以以与我们的实验室芯片技术兼容的纳升格式存储，然后用于分析。展示低温保存的纳升试剂的可行性，将允许在第二阶段开发一个完整的纳米升仪器系统，用于HTS和化学基因组学，具有前所未有的吞吐量和经济性。

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.