I-Corps: Microfluidics for Protein Crystallization and X-ray Diffraction

I-Corps项目更广泛的影响/商业潜力是为制药公司和生命科学家提供一种成本效益高、快速的方法，通过x射线晶体学来确定高分辨率的蛋白质结构。生产高分辨率的蛋白质结构是基于结构的药物设计的主要步骤之一。虽然降低成本和提高效率是基于结构的药物设计的重要好处，但缺乏高质量，快速和低成本的晶体生产的通用方法仍然是这一过程中的主要困难。微流体是一种强大的液体处理工具，可以将筛选实验中的溶液量减少到纳升，较早地被引入蛋白质结晶市场。然而，由于高性价比，这些方法都没有进入市场。我们的创新是一种用于蛋白质结晶的x射线透明微流控芯片，与目前市场上的解决方案相比，它更便宜，更容易操作，性能更高。通过将该设备商业化并推向市场，制药公司和生命科学家将减少通过x射线衍射确定蛋白质的高分辨率结构所花费的时间和金钱，这是目前基于结构的药物发现的瓶颈。这个I-Corps项目介绍了一种廉价的、x射线透明的微流控芯片，用于蛋白质结晶。对于许多医学和生物应用，如制药工程，蛋白质的分子结构是必不可少的。确定蛋白质结构最准确的方法是x射线晶体学，这一过程需要分析蛋白质晶体的x射线衍射模式。然而，蛋白质结晶仍然具有挑战性，因为每种蛋白质都有自己的相图，因此，需要尝试许多不同的条件来寻找蛋白质结晶的最佳条件。我们的技术解决了蛋白质结晶的四个核心挑战；它(1)筛选蛋白质晶体稳定性的化学条件，(2)利用优化的加工动力学产生晶体，(3)用室温晶体获得结构，(4)通过芯片上衍射避免晶体处理。片上x射线衍射已经证明了我们的蛋白质结晶芯片。我们第一个设计的原型已经制作完成，并且用一种特定的溶液进行了蛋白质结晶测试。我们的设计是一种液体处理芯片，与目前的微流控芯片相比具有多种优势。它是由廉价的生物相容性材料通过快速制造程序建造的，并且可以根据客户的需要进行修改。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 I-Corps project is to provide pharmaceutical companies and life scientists with a cost effective and fast approach for determining the high-resolution protein structure by X-ray crystallography. Production of high-resolution structures of proteins is one the main steps in structure-based drug design. Although cost reduction and efficiency improvements are the important benefits of structure-based drug design, the lack of generalized methods for high quality, fast and low-cost crystal production is still a major struggle in this process. Microfluidics, is a powerful liquid handling tool that reduces the amount of solution in a screening experiment to nanoliters, have been introduced to the protein crystallization market earlier. However, none of these approaches have penetrated the market because of the high cost to performance ratio. Our innovation is an x-ray transparent microfluidic chip for protein crystallization which is less expensive, easier to operate and more performant in comparison to the current market solutions. By commercializing this device and introducing it to the market, pharmaceutical companies and life scientists will reduce the time and money spent to determine the high-resolution structure of proteins from x-ray diffraction, which is a current bottleneck in structure-based drug discovery.This I-Corps project introduces an inexpensive, x-ray transparent microfluidic chip for protein crystallization. For many medical and biological applications, such as pharmaceutical engineering, the molecular structure of a protein is essential. The most accurate way to determine protein structure is X-ray crystallography, a process that requires the analysis of X-ray diffraction patterns of protein crystals. However, protein crystallization remains challenging because each protein has its own phase diagram, thus, many different conditions should be tried to find the optimal conditions for protein crystallization. Our technology addresses four core challenges in protein crystallization; it (1) screens chemical conditions for protein crystal stability, (2) produces crystals using optimized processing kinetics, (3) obtains structure with room temperature crystals, and (4) avoids crystal handling by diffracting on chip. On-chip X-ray diffraction has been demonstrated for our protein crystallization chips. Prototypes of our first design have been made and have been tested for protein crystallization with one specific solution. Our design is a liquid handling chip with multiple advantages over current microfluidic chips. It is built from inexpensive bio-compatible material via a rapid fabrication procedure and it can be altered as needed based on the customer need.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.