High Throughput Modular Microfluidic Systems for Drug Discovery/Development

描述(由申请人提供)：生物微机械系统(BioMEMS)被视为下一代平台，用于在药物发现等领域执行许多不同的基于生物的分析，因为它们具有提高性能和降低分析成本的潜力。然而，到目前为止，在高通量筛选(HTS)中使用生物MEMS的报道中，在产量和成本方面的潜在改进尚未实现。本研究致力于开发新的高度集成的聚合物生物微系统平台，以实现在利用并行红外荧光分析筛选候选药物文库方面的优势。为了证明该技术的有效性，将筛选L1核酸内切酶抑制剂(L1-en)。L1-EN通过TTTT‘aa共识序列诱导双链断裂，并可能导致导致细胞衰老、癌症形成或进展的遗传损伤。因此，L1-en抑制剂的发现(目前还没有发现)可以在减少癌症治疗或衰老某些方面的遗传不稳定性方面发挥重要作用。将要开发的平台有几项使能技术，这些技术将通过显著减少设备需求和最大限度地减少消耗品成本，使HTS能力传播到更广泛的药物发现社区。该系统将由大量的流体处理器(96-192)组成，这些处理器配置在使用微复制技术生产的6英寸聚合物晶片上。每个流体处理器将包括(1)互连芯片(S)，用于将化合物直接从滴定板输送到主处理器晶片；(2)微型混合器，用于加速试剂混合；(3)高表面积生物反应器，其中固定目标；以及(4)分组流体流动池，用于使用多通道荧光检测来识别潜在的铅。L1-en的底物将用新的近红外荧光酞菁染料进行双重标记，这种染料提供高灵敏度和低本底。流体流动池的平行荧光读出将通过将一组紧密间隔的通道成像到CCD相机上来进行。模拟和建模将指导选择合适的读出格式和光学配置。按照完成的格式，该系统将能够在24小时内筛选19万种候选药物。流体处理将通过微流体完成，机器人干预最少，因为只需要一个样品转移步骤就可以将候选药物加载到系统中。

DESCRIPTION (provided by applicant): BioMEMS (Biological-MicroElectroMechanical Systems) are seen as the next generation platform for performing many different biologically-based assays in areas such as drug discovery, due to their potential for providing improved performance and low assay cost. However, in reports to date of BioMEMS used in high throughput screening (HTS), the potential improvements in throughput and costs have yet to be realized. This research focuses on developing new highly integrated polymer BioMEMS platforms to achieve advantages in the screening of libraries of drug candidates using parallelized infrared fluorescence assays. As a demonstration of the efficacy of the technology, inhibitors of L1 endonuclease (L1-EN) will be screened. L1-EN induces double-strand breaks via a TTTT'AA consensus sequence and can contribute to genetic damage responsible for cellular aging, cancer formation or progression. Therefore, discovery of inhibitors of L1-EN, of which there is currently none, could play an important role in minimizing genetic instability during cancer therapies or certain aspects of aging. The platforms to be developed have several enabling technologies that will allow the dissemination of HTS capabilities into the broader drug-discovery community by significantly reducing equipment demands and minimizing consumable costs. The system will consist of a large number of fluidic processors (96-192) configured on a 6" polymer wafer produced using micro- replication. Each fluidic processor will consist of (1) interconnected chip(s) to feed compounds directly from titer plates to the main processor wafer; (2) micromixer that speeds reagent mixing; (3) high surface area bioreactor in which the target is immobilized; and (4) grouped fluidic flow-cells for identifying potential leads using multi-channel fluorescence detection. A substrate of L1-EN will be dual-labeled with new, near-IR fluorescent phthalocyanine dyes, which provide high sensitivity and low background. Parallel fluorescence readout from the fluidic flow cells will be carried out by imaging a group of tightly spaced channels onto a CCD camera. Simulation and modeling will guide the selection of an appropriate readout format and optical configuration. In its finished format, the system will be able to screen -190,000 drug candidates in 24 hr. Fluid handling will be accomplished via microfluidics with minimal robotic intervention, as only a single sample transfer step is required to load the system with drug candidates.