PFI-TT: Improved microfluidic devices for protein crystallization and x-ray diffraction

PFI-TT：改进的用于蛋白质结晶和 X 射线衍射的微流体装置

• 批准号: 1919094
• 负责人: Seth Fraden
• 金额: $ 25万
• 依托单位: Brandeis University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1919094&HistoricalAwards=false
• 关键词: PFI; TT; Improved; microfluidic; devices

The broader impact/commercial potential of this Partnerships for Innovation - Technology Translation (PFI-TT) project is in the improvement of structure-based drug design, a valuable pharmaceutical discovery tool, to accelerate discovery, reduce development costs, and boost the strength of the ultimately selected drug candidate. A critical step in this process is the accurate understanding of the three-dimensional structure of the drug target, in the presence or absence of the potential drug candidate. Currently, a method called X-ray crystallographic analysis is the most important way to determine structure, but it is expensive to operate. Two devices will be developed - one, to make small vessels to test many types of drug chemistries simultaneously, and another for better studies of the structure. Compared to current technologies, the microfluidic devices will perform better and be easier to operate, at a reduced cost.The proposed project will lead to commercialization of two microfluidic devices. The first, intended for the high-throughput screening of protein crystallization conditions, will be developed with the following features: (a) ability to screen 400 different conditions per square centimeter using counter-diffusion, (b) no cross-contamination, (c) consumes less than 10 nanoliters protein per condition, (d) low cost to manufacture and operate, and (e) improved performance over existing technologies. The technical innovation of this device is to use passive capillary valves operating with minimal external control to sequence fluid flow and create isolated storage compartments. A second device will be developed to produce crystals optimized for X-ray diffraction using a novel feedback mechanism to separate protein crystal nucleation from growth. The device will be manufactured from a low molecular weight thermoplastic, allowing on-chip serial diffraction from hundreds of identical crystals thereby avoiding expensive and time consuming cryoprotection. This will improve screening of crystallization conditions, optimize the rate of crystallization, reduce the amount of protein and eliminate laborious crystal processing steps.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该创新技术转化伙伴关系（PFI-TT）项目的更广泛影响/商业潜力在于改进基于结构的药物设计，这是一种有价值的药物发现工具，可加速发现，降低开发成本，并提高最终选定的候选药物的强度。 在这个过程中的一个关键步骤是准确理解的三维结构的药物靶标，在存在或不存在的潜在候选药物。目前，一种称为X射线晶体学分析的方法是确定结构的最重要方法，但操作成本很高。 将开发两种设备-一种是制造小血管，同时测试多种类型的药物化学物质，另一种是更好地研究结构。与现有技术相比，微流控装置将表现更好，更容易操作，成本更低。拟议的项目将导致两个微流控装置的商业化。第一种，用于蛋白质结晶条件的高通量筛选，将开发具有以下特征的：（a）使用反向扩散每平方厘米筛选400种不同条件的能力，（B）无交叉污染，（c）每种条件消耗少于10纳升蛋白质，（d）制造和操作成本低，以及（e）优于现有技术的性能。该设备的技术创新是使用被动毛细管阀，在最小的外部控制下操作，以顺序流体流动并创建隔离的储存室。第二个设备将被开发，以产生优化的X射线衍射晶体使用一种新的反馈机制，以分离蛋白质晶体成核生长。该设备将由低分子量热塑性塑料制成，允许来自数百个相同晶体的芯片上连续衍射，从而避免昂贵和耗时的低温保护。 这将改善结晶条件的筛选，优化结晶速率，减少蛋白质的量，并消除费力的晶体加工步骤。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。