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

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，可以加速发现，降低开发成本，并提高最终选定的候选药物的强度。这一过程的关键一步是在潜在候选药物存在或不存在的情况下，准确理解药物靶点的三维结构。目前，一种称为x射线晶体分析的方法是确定结构的最重要方法，但操作起来很昂贵。两种设备将被开发出来——一种是制造小血管来同时测试多种药物的化学成分，另一种是用来更好地研究药物的结构。与目前的技术相比，微流控装置性能更好，操作更容易，成本更低。拟议的项目将导致两种微流体装置的商业化。第一种用于高通量筛选蛋白质结晶条件，将具有以下特点：(a)使用反扩散筛选每平方厘米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对流体流动进行排序，并创建隔离的存储室。第二种装置将被开发出来，使用一种新的反馈机制来分离蛋白质晶体的成核和生长，从而生产出适合x射线衍射的晶体。该设备将由低分子量热塑性塑料制成，允许数百个相同晶体在片上连续衍射，从而避免昂贵和耗时的冷冻保护。这将改善结晶条件的筛选，优化结晶速率，减少蛋白质的用量，消除繁琐的结晶处理步骤。该奖项反映了美国国家科学基金会的法定使命，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。

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.